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Phospho-IKB alpha (Ser32) Antibody (YA183)

(Synonyms: IKBA, MAD3, NFKBI, NFKBIA, NF-kappa-B inhibitor alpha, I-kappa-B-alpha, Major histocompatibility complex enhancer-binding protein MAD3, IkB-alpha, IkappaBalpha)
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Based on 1 Customer Validation

Phospho-IKB alpha (Ser32) Antibody (YA183) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to Phospho-IKB alpha (Ser32).

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

    Rabbit

  • Isotype:

    IgG

  • Application:

    WB, ICC/IF, IP

  • Reactivity :

    Human, Mouse

  • Formulation:

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

  • Conjugation:
    Non-conjugated
  • Experimental Validation Results for Phospho-IKB alpha (Ser32) Antibody (YA183)
  • Experimental Validation Results for Phospho-IKB alpha (Ser32) Antibody (YA183)
  • Phospho-IKB alpha (Ser32) Antibody (YA183) Product Image
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Tel: 609-228-6898 E-mail: [email protected] [email protected]

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Use of MCE Products

Applications

Application
WB Info
WB: Western Blot
ICC/IF Info
ICC/IF: Immunocytochemistry/
Immunofluorescence
IP Info
IP: Immunoprecipitation
Dilution Ratio 1:1000 1:50-1:200 Use at an assay dependent concentration.

Product Details

Description

Phospho-IKB alpha (Ser32) Antibody (YA183) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to Phospho-IKB alpha (Ser32).

  • Host Rabbit
  • Clonality Recombinant,Monoclonal
  • Species Reactivity
    Human, Mouse
  • Observed Molecular Weight
    Observed band size: 36 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: 36 kDa
Species Reactivity Database
Immunogen

Synthetic phosphopeptide corresponding to residues surrounding Ser32 of Human IKB alpha.The exact sequence is proprietary to MCE.

Sensitivity

Endogenous

Purification

Protein A affinity purified.

Conjugation

Non-conjugated

Modification

Phosphorylated

Isotype

IgG

RRID

AB_3102540

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-IKB alpha (Ser32) Antibody (YA183)
    Western blot analysis of extracts from Hela(lane 2(20μg) ) and Hela(lane 3(20μg), Human TNF-alpha treated (HY-P7058, 20 ng/mL Human TNF-α, 2hrs) as indicated, using Phospho-IKB alpha (Ser32) (HY-P80446) Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% BSA 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% BSA in TBST at 4°C overnight. Goat Anti-Rabbit/Mouse IgG-HRP Secondary Antibody (1/10,000) was used for 1 hour at room temperature.
  • Experimental Validation Results for Phospho-IKB alpha (Ser32) Antibody (YA183)
    Immunocytochemistry analysis of Hela cells labeling Phospho-IKB alpha (Ser32) with Phospho-IKB alpha (Ser32) antibody (HY-P80466) 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-IKB alpha (Ser32) antibody (HY-P80466) 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).

Background

  • Function

    IKB alpha inhibits the activity of dimeric NF-kappa-B/REL complexes by trapping REL (RELA/p65 and NFKB1/p50) dimers in the cytoplasm by masking their nuclear localization signals. On cellular stimulation by immune and pro-inflammatory responses, becomes phosphorylated promoting ubiquitination and degradation, enabling the dimeric RELA to translocate to the nucleus and activate transcription[1][2][3][4][5][6].

  • Subcellular Localization

    Cytoplasm; Nucleus

  • Expression


    Induction:Induced in adherent monocytes

  • Subunit

    Interacts with RELA; the interaction requires the nuclear import signal (PubMed:1493333). Part of a 70-90 kDa complex at least consisting of CHUK, IKBKB, NFKBIA, RELA, ELP1 and MAP3K14 (PubMed:9751059). Interacts with NKIRAS1 and NKIRAS2 (PubMed:10657303). Interacts with isoform 1 and isoform 2 of RWDD3; the interaction enhances sumoylation (PubMed:17956732, PubMed:23469069). Interacts with PRMT2 (PubMed:16648481). Interacts with PRKACA in platelets; this interaction is disrupted by thrombin and collagen (PubMed:20356841). Interacts with MEFV (PubMed:18577712). Interacts with DDRGK1; positively regulates NFKBIA phosphorylation and degradation (PubMed:23675531). Interacts with HNRNPA2B1; the interaction may be mediated by the RRM2 domain of HNRNPA2B1, and HNRNPA2B1 may interact simultaneously with FAM76B and either NFKBIA or NFKBIE to form a complex (PubMed:37643469)

  • SwissProt ID

    P25963

  • Gene ID
    4792 [NCBI]
  • Synonyms

    IKBA, MAD3, NFKBI, NFKBIA, NF-kappa-B inhibitor alpha, I-kappa-B-alpha, Major histocompatibility complex enhancer-binding protein MAD3, IkB-alpha, IkappaBalpha

  • Research Field

    Signal Transduction

References

[1]. Ganchi PA, et al. I kappa B/MAD-3 masks the nuclear localization signal of NF-kappa B p65 and requires the transactivation domain to inhibit NF-kappa B p65 DNA binding. Mol Biol Cell. 1992 Dec;3(12):1339-52. [Content Brief]

[2]. Quan J, et al. USP39 Regulates NF-κB-Mediated Inflammatory Responses through Deubiquitinating K48-Linked IκBα. J Immunol. 2023 Mar 1;210(5):640-652. [Content Brief]

[3]. Scherer DC, et al. Signal-induced degradation of I kappa B alpha requires site-specific ubiquitination. Proc Natl Acad Sci U S A. 1995 Nov 21;92(24):11259-63. [Content Brief]

[4]. Chen Z, et al. Signal-induced site-specific phosphorylation targets I kappa B alpha to the ubiquitin-proteasome pathway. Genes Dev. 1995 Jul 1;9(13):1586-97. [Content Brief]

[5]. Traenckner EB, et al. Phosphorylation of human I kappa B-alpha on serines 32 and 36 controls I kappa B-alpha proteolysis and NF-kappa B activation in response to diverse stimuli. EMBO J. 1995 Jun 15;14(12):2876-83. [Content Brief]

[6]. Brown K, et al. Control of I kappa B-alpha proteolysis by site-specific, signal-induced phosphorylation. Science. 1995 Mar 10;267(5203):1485-8. [Content Brief]

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Phospho-IKB alpha (Ser32) Antibody (YA183) 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 Hela(lane 2(20μg) ) and Hela(lane 3(20μg), Human TNF-alpha treated (HY-P7058, 20 ng/mL Human TNF-α, 2hrs) as indicated, using Phospho-IKB alpha (Ser32) (HY-P80446) Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% BSA 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% BSA in TBST at 4°C overnight. Goat Anti-Rabbit/Mouse IgG-HRP Secondary Antibody (1/10,000) was used for 1 hour at room temperature.

  • 1/16

    MedchemExpress Validation
     Immunocytochemistry analysis of Hela cells labeling Phospho-IKB alpha (Ser32) with Phospho-IKB alpha (Ser32) antibody (HY-P80466) 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-IKB alpha (Ser32) antibody (HY-P80466) 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-IKB alpha (Ser32) Antibody (YA183) Product Image
    1/16
  • Experimental Validation Results for Phospho-IKB alpha (Ser32) Antibody (YA183)
  • Experimental Validation Results for Phospho-IKB alpha (Ser32) Antibody (YA183)
  • Phospho-IKB alpha (Ser32) Antibody (YA183) Product Image
  • Experimental Validation Results for Phospho-IKB alpha (Ser32) Antibody (YA183)
    1/1
    MedchemExpress Validation
     Western blot analysis of extracts from Hela(lane 2(20μg) ) and Hela(lane 3(20μg), Human TNF-alpha treated (HY-P7058, 20 ng/mL Human TNF-α, 2hrs) as indicated, using Phospho-IKB alpha (Ser32) (HY-P80446) Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% BSA 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% BSA in TBST at 4°C overnight. Goat Anti-Rabbit/Mouse IgG-HRP Secondary Antibody (1/10,000) was used for 1 hour at room temperature.
  • Experimental Validation Results for Phospho-IKB alpha (Ser32) Antibody (YA183)
    1/1
    MedchemExpress Validation
     Immunocytochemistry analysis of Hela cells labeling Phospho-IKB alpha (Ser32) with Phospho-IKB alpha (Ser32) antibody (HY-P80466) 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-IKB alpha (Ser32) antibody (HY-P80466) 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-IKB alpha (Ser32) Antibody (YA183)
  • Experimental Validation Results for Phospho-IKB alpha (Ser32) Antibody (YA183)
  • Phospho-IKB alpha (Ser32) Antibody (YA183) Product Image
    MedchemExpress Validation

    Western blot analysis of extracts from Hela(lane 2(20μg) ) and Hela(lane 3(20μg), Human TNF-alpha treated (HY-P7058, 20 ng/mL Human TNF-α, 2hrs) as indicated, using Phospho-IKB alpha (Ser32) (HY-P80446) Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% BSA 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% BSA in TBST at 4°C overnight. Goat Anti-Rabbit/Mouse IgG-HRP Secondary Antibody (1/10,000) was used for 1 hour at room temperature.

  • Phospho-IKB alpha (Ser32) Antibody (YA183) Product Image
    MedchemExpress Validation

    Immunocytochemistry analysis of Hela cells labeling Phospho-IKB alpha (Ser32) with Phospho-IKB alpha (Ser32) antibody (HY-P80466) 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-IKB alpha (Ser32) antibody (HY-P80466) 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-IKB alpha (Ser32) Antibody (YA183) Product Image WB
  • Phospho-IKB alpha (Ser32) Antibody (YA183) Product Image ICC