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Phospho-AKT1(Ser473) Antibody (YA227)

(Synonyms: AKT 1 antibody; AKT antibody; AKT1 antibody; AKT1_HUMAN antibody; MGC99656 antibody; PKB antibody; PKB-ALPHA antibody; PRKBA antibody; Protein Kinase B Alpha antibody; Protein kinase B antibody; Proto-oncogene c-Akt antibody; RAC Alpha antibody; RAC antibody; RAC-alpha serine/threonine-protein kinase antibody; RAC-PK-alpha antibody; )
4 Cited Publications
Customer Review

Based on 4 publication(s) in Google Scholar

Phospho-AKT1(Ser473) Antibody (YA227) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to Phospho-AKT1(Ser473).

For research use only. We do not sell to patients.
  • Host:

    Rabbit

  • Isotype:

    IgG

  • Application:

    WB, ICC/IF, IHC-P

  • Reactivity :

    Human, Mouse, Rat, Dog

  • Formulation:

    Supplied in 1*TBS (pH7.4), 0.05% BSA and 40% Glycerol. Preservative: 0.05% Sodium Azide.

  • Conjugation:
    Non-conjugated
ALL WB ICC IHC-P
  • Experimental Validation Results for Phospho-AKT1(Ser473) Antibody (YA227)
  • Experimental Validation Results for Phospho-AKT1(Ser473) Antibody (YA227)
  • Experimental Validation Results for Phospho-AKT1(Ser473) Antibody (YA227)
  • Experimental Validation Results for Phospho-AKT1(Ser473) Antibody (YA227)
  • Experimental Validation Results for Phospho-AKT1(Ser473) Antibody (YA227)
  • Phospho-AKT1(Ser473) Antibody (YA227) Product Image
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Publications Citing Use of MedChemExpress (MCE) Phospho-AKT1(Ser473) Antibody (YA227)

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4 Publications Citing Use of MCE Phospho-AKT1(Ser473) Antibody (YA227) (8)
WB
  • WB
  • WB
  • WB
  • WB
  • WB
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  • WB
  • WB

Applications

Application
WB Info
WB: Western Blot
ICC/IF Info
ICC/IF: Immunocytochemistry/
Immunofluorescence
IHC-P Info
IHC-P: Immunohistochemistry-Paraffin
IF-Tissue Info
IF-Tissue: Immunofluorescence-Tissue
IHC-F Info
IHC-F: Immunohistochemistry-Frozen
Dilution Ratio 1:5000-1:10000 1:100-1:1000 1:200-1:1000 1:200-1:500 1:100

Product Details

Description

Phospho-AKT1(Ser473) Antibody (YA227) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to Phospho-AKT1(Ser473).

  • Host Rabbit
  • Clonality Recombinant,Monoclonal
  • Species Reactivity
    Human, Mouse, Rat, Dog
  • Observed Molecular Weight
    Observed band size: 56 kDa Info
    Note: Due to possible protein modifications or aggregation, the molecular weight should be confirmed by actual measurement, and the predicted value is for reference only.
  • Calculated Molecular Weight Predicted band size: 56 kDa
Species Reactivity Database
Immunogen

Synthetic phosphopeptide corresponding to residues surrounding Ser473 of Human Akt1.The exact sequence is proprietary to MCE.

Sensitivity

Endogenous

Purification

Protein A affinity purified.

Conjugation

Non-conjugated

Modification

Phosphorylated

Isotype

IgG

RRID

AB_3102397

Product Properties

  • Appearance

    Solution

  • Formulation

    Supplied in 1*TBS (pH7.4), 0.05% BSA and 40% Glycerol. Preservative: 0.05% Sodium Azide.

  • Concentration

    Batch-dependent, Please check the COA for the concentration of each lot. Check Lot Concentration

  • Storage & Stability

    Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.

  • Shipping

    Shipping with blue ice.

Verification Images

  • Experimental Validation Results for Phospho-AKT1(Ser473) Antibody (YA227)
    Western blot analysis of extracts from A549(lane 2(20μg) , MCF-7(lane 3(20μg) and Jurkat(lane 4(20ug) using Phospho-AKT1(HY-P80276 Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% non-fat milk in TBST for 2 hour at room temperature. The primary antibody (1/1000) and Loading control antibody (Beta Actin, HY-P80438, 1/10000) was used in 5% non-fat milk in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (1/10000) was used for 1 hour at room temperature.
  • Experimental Validation Results for Phospho-AKT1(Ser473) Antibody (YA227)
    Immunocytochemistry analysis of Hela cells treated with or without 100nM Calyculin A for 30 minutes labeling Phospho-AKT1(Ser473) with Phospho-AKT1(Ser473) Antibody (HY-P80276)at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with Phospho-AKT1(Ser473) Antibody (HY-P80276) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
  • Experimental Validation Results for Phospho-AKT1(Ser473) Antibody (YA227)
    Immunocytochemistry analysis of C6 cells treated with or without 100nM Calyculin A for 30 minutes labeling Phospho-AKT1(Ser473) with A Phospho-AKT1(Ser473) Antibody (HY-P80276) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with Phospho-AKT1(Ser473) Antibody (HY-P80276)at 1/100 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
  • Experimental Validation Results for Phospho-AKT1(Ser473) Antibody (YA227)
    Immunohistochemical analysis of paraffin-embedded mouse lung tissue using Phospho-AKT1 Antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/100 dilution in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Experimental Validation Results for Phospho-AKT1(Ser473) Antibody (YA227)
    Immunohistochemical analysis of paraffin-embedded mouse lung tissue using Phospho-AKT1 Antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/100 dilution in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Background

  • Function

    AKT1 (protein kinase Bα, PKBα) is a serine/threonine kinase that functions as a central effector of the PI3K/AKT/mTOR signaling pathway, regulating cellular growth, survival, metabolism, proliferation, and differentiation across multiple tissues and cell types[1][2][3]. Activation of AKT1 occurs downstream of PI3K through phosphoinositide-dependent kinase-1 (PDK1) -mediated phosphorylation, enabling downstream regulation of targets involved in protein synthesis, apoptosis control, and metabolic adaptation[4][5]. Mechanistically, AKT1 contributes to angiogenesis, physiological tissue growth, immune-cell function, and stress-response signaling, making it a critical node in both normal physiology and disease-associated signaling networks[2][4][5]. In disease models, dysregulated AKT signaling is strongly associated with cancer, metabolic disorders, cardiovascular pathology, and inflammatory conditions, and constitutive activation of AKT-dependent pathways promotes oncogenic progression and therapeutic resistance[2][6][7]. Compared with related isoforms, AKT1 exhibits distinct biological functions despite substantial structural homology with AKT2 and AKT3; genetic studies demonstrate that AKT1 deficiency primarily causes growth retardation, whereas AKT2 deficiency induces insulin resistance and diabetes, and AKT3 loss predominantly affects brain development[4][5]. Isoform-specific investigations further show that AKT1 and AKT2 can exert non-redundant or even opposing effects on migration, invasion, inflammation, and fibrogenesis depending on cellular context[2][6][8]. For experimental applications, increasing efforts focus on developing AKT1-selective inhibitors and isoform-selective cellular models, which provide valuable tools for dissecting AKT isoform-specific signaling and for evaluating targeted therapeutic strategies with improved selectivity profiles[6][7][9].

  • Subcellular Localization

    Cytoplasm; Nucleus; Cell membrane; Mitochondrion intermembrane space

  • Expression


    Tissue_specificity:This protein is expressed in prostate cancer, and its expression level gradually increases from normal to malignant (protein level) . It is expressed in all human cell types analyzed to date. The phosphorylated form of Tyr-176 is significantly increased in the progressive stages of breast cancer (i.e., from normal to hyperplastic (ADH) , ductal carcinoma in situ (DCIS) , invasive ductal carcinoma (IDC) , and lymph node metastasis (LNMM) stages) .

  • Subunit

    Interacts with BTBD10 (By similarity). Interacts with KCTD20 (By similarity). Interacts (via the C-terminus) with CCDC88A (via its C-terminus). Interacts with GRB10; the interaction leads to GRB10 phosphorylation thus promoting YWHAE-binding (By similarity). Interacts with AGAP2 (isoform 2/PIKE-A); the interaction occurs in the presence of guanine nucleotides. Interacts with AKTIP. Interacts (via PH domain) with MTCP1, TCL1A and TCL1B. Interacts with CDKN1B; the interaction phosphorylates CDKN1B promoting 14-3-3 binding and cell-cycle progression. Interacts with MAP3K5 and TRAF6. Interacts with BAD, PPP2R5B, STK3 and STK4. Interacts (via PH domain) with SIRT1. Interacts with SRPK2 in a phosphorylation-dependent manner. Interacts with RAF1. Interacts with TRIM13; the interaction ubiquitinates AKT1 leading to its proteasomal degradation. Interacts with TNK2 and CLK2. Interacts (via the C-terminus) with THEM4 (via its C-terminus). Interacts with and phosphorylated by PDPK1. Interacts with PA2G4 (By similarity). Interacts with KIF14; the interaction is detected in the plasma membrane upon INS stimulation and promotes AKT1 phosphorylation (PubMed:24784001). Interacts with FAM83B; activates the PI3K/AKT signaling cascade (PubMed:23676467). Interacts with WDFY2 (via WD repeats 1-3) (PubMed:16792529). Forms a complex with WDFY2 and FOXO1 (By similarity). Interacts with FAM168A (PubMed:23251525). Interacts with SYAP1 (via phosphorylated form and BSD domain); this interaction is enhanced in a mTORC2-mediated manner in response to epidermal growth factor (EGF) stimulation and activates AKT1 (PubMed:23300339). Interacts with PKHM3 (By similarity). Interacts with FKBP5/FKBP51; promoting interaction between Akt/AKT1 and PHLPP1, thereby enhancing dephosphorylation and subsequent activation of Akt/AKT1 (PubMed:28147277). Interacts with TMEM175; leading to formation of the lysoK(GF) complex (PubMed:32228865). Acts as a negative regulator of the cGAS-STING pathway by mediating phosphorylation of CGAS during mitosis, leading to its inhibition (PubMed:26440888)

  • SwissProt ID

    P31749

  • Gene ID
    207 [NCBI]
  • Synonyms

    AKT 1 antibody; AKT antibody; AKT1 antibody; AKT1_HUMAN antibody; MGC99656 antibody; PKB antibody; PKB-ALPHA antibody; PRKBA antibody; Protein Kinase B Alpha antibody; Protein kinase B antibody; Proto-oncogene c-Akt antibody; RAC Alpha antibody; RAC antibody; RAC-alpha serine/threonine-protein kinase antibody; RAC-PK-alpha antibody;

  • Research Field

    Neuroscience

References

[1]. Guerau-de-Arellano M, et al. Akt isoforms in the immune system. Front Immunol. 2022 Aug 23;13:990874. [Content Brief]

[2]. Reyes-Gordillo K, et al. Akt1 and Akt2 Isoforms Play Distinct Roles in Regulating the Development of Inflammation and Fibrosis Associated with Alcoholic Liver Disease. Cells. 2019 Oct 29;8(11):1337. [Content Brief]

[3]. O'Neill BT, et al. Akt1 in the cardiovascular system: friend or foe? J Clin Invest. 2005 Aug;115(8):2059-64. [Content Brief]

[4]. Tellai AD, et al. The complex post-transcriptional regulation of genes coding for methionine adenosyl transferase: New insights for liver cancer. Biochimie. 2025 Nov;238(Pt A):19-28. [Content Brief]

[5]. Quambusch L, et al. Cellular model system to dissect the isoform-selectivity of Akt inhibitors. Nat Commun. 2021 Sep 6;12(1):5297. [Content Brief]

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Phospho-AKT1(Ser473) Antibody (YA227) Related Classifications

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Measured by its ability to inhibit anti-CD3-induced proliferation of stimulated CTLL-2 mouse cytotoxic T cells. The ED50 this effect is 0.8023 μg/ml, corresponding to a specific activity is 1246.42 units/mg.

Assay Types
  • 1/16

    MedChemExpress Validation
     Western blot analysis of extracts from A549(lane 2(20μg) , MCF-7(lane 3(20μg) and Jurkat(lane 4(20ug) using Phospho-AKT1(HY-P80276 Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% non-fat milk in TBST for 2 hour at room temperature. The primary antibody (1/1000) and Loading control antibody (Beta Actin, HY-P80438, 1/10000) was used in 5% non-fat milk in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (1/10000) was used for 1 hour at room temperature.

  • 1/16

    MedChemExpress Validation
     Immunocytochemistry analysis of Hela cells treated with or without 100nM Calyculin A for 30 minutes labeling Phospho-AKT1(Ser473) with Phospho-AKT1(Ser473) Antibody (HY-P80276)at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with Phospho-AKT1(Ser473) Antibody (HY-P80276) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).

  • 1/16

    MedChemExpress Validation
     Immunocytochemistry analysis of C6 cells treated with or without 100nM Calyculin A for 30 minutes labeling Phospho-AKT1(Ser473) with A Phospho-AKT1(Ser473) Antibody (HY-P80276) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with Phospho-AKT1(Ser473) Antibody (HY-P80276)at 1/100 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).

  • 1/16

    MedChemExpress Validation
     Immunohistochemical analysis of paraffin-embedded mouse lung tissue using Phospho-AKT1 Antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/100 dilution in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • 1/16

    MedChemExpress Validation
     Immunohistochemical analysis of paraffin-embedded mouse lung tissue using Phospho-AKT1 Antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/100 dilution in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • 1/16

    Phospho-AKT1(Ser473) Antibody (YA227) purchased from MedChemExpress. Usage Cited in: Phytomedicine. 2026 Jan:150:157741. [Abstract]

    Customer Validation
     A549 and LLC cells were treated with increasing doses of TXC for 24 h, followed by western blotting for ERBB2, PI3K, phospho-PI3K, AKT, phospho-AKT, mTOR, phospho-Mtor.

  • 1/16

    Phospho-AKT1(Ser473) Antibody (YA227) purchased from MedChemExpress. Usage Cited in: Phytomedicine. 2026 Jan:150:157741. [Abstract]

    Customer Validation
     Subcutaneous LLC xenografts were established in C57BL/6 mice, the tumors were harvested on day 18. Western blot analysis of ERBB2, PI3K, phospho-PI3K, AKT, phospho-AKT, mTOR, and phospho-mTOR in tumor lysates.

  • 1/16

    Phospho-AKT1(Ser473) Antibody (YA227) purchased from MedChemExpress. Usage Cited in: Phytomedicine. 2026 Jan:150:157741. [Abstract]

    Customer Validation
     Western blot analysis of ERBB2, PI3K, phospho-PI3K, AKT, phospho-AKT, mTOR, phospho-mTOR, HMOX1, and GPX4 in HCC827 and HCC827-GR cells treated with TXC or gefitinib.

  • 1/16

    Phospho-AKT1(Ser473) Antibody (YA227) purchased from MedChemExpress. Usage Cited in: Phytomedicine. 2026 Jan:150:157741. [Abstract]

    Customer Validation
     Following 12 h of cotreatment, Western blotting for ERBB2, PI3K, phospho-PI3K, AKT, phospho-AKT, mTOR, phospho-mTOR, HMOX1, GPX4, P-gp, and MRP1.

  • 1/16

    Phospho-AKT1(Ser473) Antibody (YA227) purchased from MedChemExpress. Usage Cited in: Neuropharmacology. 2026 Jul 1:292:110950. [Abstract]

    Customer Validation
     The expressions of p-PI3K, PI3K, p-AKT, AKT, and ACSL4 in the SN of mice brain were detected by Western blot.

  • 1/16

    Phospho-AKT1(Ser473) Antibody (YA227) purchased from MedChemExpress. Usage Cited in: Neuropharmacology. 2026 Jul 1:292:110950. [Abstract]

    Customer Validation
     SH-SY5Y and PC12 cells were treated with MPP+ (1 mM) in the presence of YL-0919 (10 μg/mL) for 24 h, and expressions of p-PI3K, PI3K, p-AKT, AKT, and ACSL4 were detected by Western blot.

  • 1/16

    Phospho-AKT1(Ser473) Antibody (YA227) purchased from MedChemExpress. Usage Cited in: J Inflamm Res. 2025 Apr 28:18:5727-5739. [Abstract]

    Customer Validation
     THP-1 cells were infected with M. pneumoniae at MOIs of 0, 10, 50, or 100 for 1 h, and protein expression levels were assessed by Western blot.

  • 1/16

    Phospho-AKT1(Ser473) Antibody (YA227) purchased from MedChemExpress. Usage Cited in: Viruses. 2025 Dec 17;17(12):1637. [Abstract]

    Customer Validation
     Quercetin suppresses the PI3K/Akt/mTOR signaling pathway. MARC-145 cells were infected with PRRSV (MOI of 0.8) and treated with quercetin for 48 h.

  • Phospho-AKT1(Ser473) Antibody (YA227) Product Image
    1/16
  • Experimental Validation Results for Phospho-AKT1(Ser473) Antibody (YA227)
  • Experimental Validation Results for Phospho-AKT1(Ser473) Antibody (YA227)
  • Experimental Validation Results for Phospho-AKT1(Ser473) Antibody (YA227)
  • Experimental Validation Results for Phospho-AKT1(Ser473) Antibody (YA227)
  • Experimental Validation Results for Phospho-AKT1(Ser473) Antibody (YA227)
  • Experimental Validation Results for Phospho-AKT1(Ser473) Antibody (YA227)
  • Experimental Validation Results for Phospho-AKT1(Ser473) Antibody (YA227)
  • Experimental Validation Results for Phospho-AKT1(Ser473) Antibody (YA227)
  • Experimental Validation Results for Phospho-AKT1(Ser473) Antibody (YA227)
  • Experimental Validation Results for Phospho-AKT1(Ser473) Antibody (YA227)
  • Experimental Validation Results for Phospho-AKT1(Ser473) Antibody (YA227)
  • Experimental Validation Results for Phospho-AKT1(Ser473) Antibody (YA227)
  • Experimental Validation Results for Phospho-AKT1(Ser473) Antibody (YA227)
  • Phospho-AKT1(Ser473) Antibody (YA227) Product Image
  • Experimental Validation Results for Phospho-AKT1(Ser473) Antibody (YA227)
    1/1
    MedChemExpress Validation
     Western blot analysis of extracts from A549(lane 2(20μg) , MCF-7(lane 3(20μg) and Jurkat(lane 4(20ug) using Phospho-AKT1(HY-P80276 Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% non-fat milk in TBST for 2 hour at room temperature. The primary antibody (1/1000) and Loading control antibody (Beta Actin, HY-P80438, 1/10000) was used in 5% non-fat milk in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (1/10000) was used for 1 hour at room temperature.
  • Experimental Validation Results for Phospho-AKT1(Ser473) Antibody (YA227)
    1/1
    MedChemExpress Validation
     Immunocytochemistry analysis of Hela cells treated with or without 100nM Calyculin A for 30 minutes labeling Phospho-AKT1(Ser473) with Phospho-AKT1(Ser473) Antibody (HY-P80276)at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with Phospho-AKT1(Ser473) Antibody (HY-P80276) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
  • Experimental Validation Results for Phospho-AKT1(Ser473) Antibody (YA227)
    1/1
    MedChemExpress Validation
     Immunocytochemistry analysis of C6 cells treated with or without 100nM Calyculin A for 30 minutes labeling Phospho-AKT1(Ser473) with A Phospho-AKT1(Ser473) Antibody (HY-P80276) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with Phospho-AKT1(Ser473) Antibody (HY-P80276)at 1/100 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
  • Experimental Validation Results for Phospho-AKT1(Ser473) Antibody (YA227)
    1/1
    MedChemExpress Validation
     Immunohistochemical analysis of paraffin-embedded mouse lung tissue using Phospho-AKT1 Antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/100 dilution in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Experimental Validation Results for Phospho-AKT1(Ser473) Antibody (YA227)
    1/1
    MedChemExpress Validation
     Immunohistochemical analysis of paraffin-embedded mouse lung tissue using Phospho-AKT1 Antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/100 dilution in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Experimental Validation Results for Phospho-AKT1(Ser473) Antibody (YA227)
  • Experimental Validation Results for Phospho-AKT1(Ser473) Antibody (YA227)
  • Experimental Validation Results for Phospho-AKT1(Ser473) Antibody (YA227)
  • Experimental Validation Results for Phospho-AKT1(Ser473) Antibody (YA227)
  • Experimental Validation Results for Phospho-AKT1(Ser473) Antibody (YA227)
  • Phospho-AKT1(Ser473) Antibody (YA227) Product Image
    MedChemExpress Validation

    Western blot analysis of extracts from A549(lane 2(20μg) , MCF-7(lane 3(20μg) and Jurkat(lane 4(20ug) using Phospho-AKT1(HY-P80276 Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% non-fat milk in TBST for 2 hour at room temperature. The primary antibody (1/1000) and Loading control antibody (Beta Actin, HY-P80438, 1/10000) was used in 5% non-fat milk in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (1/10000) was used for 1 hour at room temperature.

  • Phospho-AKT1(Ser473) Antibody (YA227) Product Image
    MedChemExpress Validation

    Immunocytochemistry analysis of Hela cells treated with or without 100nM Calyculin A for 30 minutes labeling Phospho-AKT1(Ser473) with Phospho-AKT1(Ser473) Antibody (HY-P80276)at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with Phospho-AKT1(Ser473) Antibody (HY-P80276) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).

  • Phospho-AKT1(Ser473) Antibody (YA227) Product Image
    MedChemExpress Validation

    Immunocytochemistry analysis of C6 cells treated with or without 100nM Calyculin A for 30 minutes labeling Phospho-AKT1(Ser473) with A Phospho-AKT1(Ser473) Antibody (HY-P80276) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with Phospho-AKT1(Ser473) Antibody (HY-P80276)at 1/100 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).

  • Phospho-AKT1(Ser473) Antibody (YA227) Product Image
    MedChemExpress Validation

    Immunohistochemical analysis of paraffin-embedded mouse lung tissue using Phospho-AKT1 Antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/100 dilution in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Phospho-AKT1(Ser473) Antibody (YA227) Product Image
    MedChemExpress Validation

    Immunohistochemical analysis of paraffin-embedded mouse lung tissue using Phospho-AKT1 Antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/100 dilution in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Phospho-AKT1(Ser473) Antibody (YA227) Product Image WB
  • Phospho-AKT1(Ser473) Antibody (YA227) Product Image ICC
  • Phospho-AKT1(Ser473) Antibody (YA227) Product Image ICC
  • Phospho-AKT1(Ser473) Antibody (YA227) Product Image IHC-P
  • Phospho-AKT1(Ser473) Antibody (YA227) Product Image IHC-P