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Smad2/3 Antibody

(Synonyms: SMAD2; MADH2; MADR2; Mothers against decapentaplegic homolog 2; MAD homolog 2; Mothers against DPP homolog 2; JV18-1; Mad-related protein 2; hMAD-2; SMAD family member 2; SMAD 2; Smad2; hSMAD2)
Customer Review

Based on 1 Customer Validation

Smad2/3 Antibody is a Rabbit-derived and non-conjugated IgG polyclonal antibody, targeting to Smad2/3.

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

    Rabbit

  • Isotype:

    IgG

  • Application:

    WB, ICC/IF, FC, IP, ChIP

  • Reactivity :

    Human, Mouse, Rat

  • Formulation:

    Supplied in 50 mM Tris-Glycine (pH 7.4), 0.15 M NaCl, 40% Glycerol and 0.05% BSA. Preservative: 0.01% Sodium azide

  • Conjugation:
    Non-conjugated
ALL WB FC ICC
  • Experimental Validation Results for Smad2/3 Antibody
  • Experimental Validation Results for Smad2/3 Antibody
  • Experimental Validation Results for Smad2/3 Antibody
  • Experimental Validation Results for Smad2/3 Antibody
  • Experimental Validation Results for Smad2/3 Antibody
  • Smad2/3 Antibody Product Image
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To place orders, for customer services and technical support, please contact: MedChemExpress USA

Tel: 609-228-6898 E-mail: [email protected] [email protected]

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Applications

Application
WB Info
WB: Western Blot
ICC/IF Info
ICC/IF: Immunocytochemistry/
Immunofluorescence
IP Info
IP: Immunoprecipitation
FC Info
FC: Flow Cytometry
ChIP Info
ChIP: Chromatin Immunoprecipitation
Dilution Ratio 1:500-1:1000 1:50-1:200 1:20 1:50-1:100 1:20

Product Details

Description

Smad2/3 Antibody is a Rabbit-derived and non-conjugated IgG polyclonal antibody, targeting to Smad2/3.

  • Host Rabbit
  • Clonality Polyclonal
  • Species Reactivity
    Human, Mouse, Rat
  • Observed Molecular Weight
    Observed band size: 52,60 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: 52 kDa
Species Reactivity Database
Immunogen

Synthetic peptide corresponding to Human Smad2 smad2 aa2-280.

Sensitivity

Endogenous

Purification

affinity purified

Conjugation

Non-conjugated

Modification

Unmodified

Isotype

IgG

RRID

AB_3102740

Product Properties

  • Appearance

    Solution

  • Formulation

    Supplied in 50 mM Tris-Glycine (pH 7.4), 0.15 M NaCl, 40% Glycerol and 0.05% BSA. Preservative: 0.01% 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 Smad2/3 Antibody
    Western blot analysis of extracts from Hela(lane 2(20ug) , HEK293(lane 3(20ug) and C6(lane 4(20ug) using Smad2/3 Antibody (HY-P80893) 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-P83730, 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 Smad2/3 Antibody
    Western blot analysis was performed on extracts from Hela (lane 1, 15 μg), HT-1080 (lane 2, 15 μg), A549 (lane 3, 15 μg), LnCaP (lane 4, 15 μg), RAW264.7 (lane 5, 15 μg), and C6 (lane 6, 15 μg) using Smad2/3 Rabbit pAb. Proteins were transferred to a PVDF membrane and blocked with 5% non - fat milk in TBST at 4°C overnight. The primary antibody (1:1000 dilution) and the loading control antibody (beta-Actin(HRP), HY-P80993, 1:5000 dilution) was incubated in 5% non-fat milk in TBST for 1 hour at 37°C. Goat Anti - Rabbit IgG - HRP Secondary Antibody (1:20000 dilution) was then applied for 40 minutes at 37°C.
  • Experimental Validation Results for Smad2/3 Antibody
    Flow cytometric analysis of 1X106 NIH-3T3 cells labeling Smad2/3 Antibody(HY-P80893, red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Then stained with the primary antibody at 1/100 dilution for an hour at 4℃. AF488-conjugated Goat Anti-Rabbit IgG H&L (HY-P8002) was used as the secondary antibody at 1/1,000 dilution for 30 minutes at 4℃. Rabbit IgG Isotype Control (HY-P80879, blue) was used as the isotype control, cells without incubation with primary antibody were used as the unlabeled control (black).
  • Experimental Validation Results for Smad2/3 Antibody
    Immunocytochemistry analysis of Hela cells labeling Smad2/3 Antibody (HY-P80893) 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 Smad2/3 Antibody (HY-P80893) 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 Smad2/3 Antibody
    Immunocytochemistry analysis of A549 cells labeling Smad2/3 Antibody (HY-P80893) 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 Smad2/3 Antibody (HY-P80893) 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

    Smad2/3 is a Receptor-regulated SMAD (R-SMAD) that is an intracellular signal transducer and transcriptional modulator activated by TGF-beta (transforming growth factor) and activin type 1 receptor kinases. Binds the TRE element in the promoter region of many genes that are regulated by TGF-beta and, on formation of the SMAD2/SMAD4 complex, activates transcription. Promotes TGFB1-mediated transcription of odontoblastic differentiation genes in dental papilla cells. Positively regulates PDPK1 kinase activity by stimulating its dissociation from the 14-3-3 protein YWHAQ which acts as a negative regulator. May act as a tumor suppressor in colorectal carcinoma[1].

  • Subcellular Localization

    Cytoplasm; Nucleus

  • Expression


    Tissue_specificity:Highly expressed in skeletal muscle, endothelial cells, heart, and placenta.

  • Isoforms & Post-Translational Modification

    Q15796 has 2 isomers: Q15796-1: 52306 Da (predicted); Q15796-2: 48956 Da (predicted).
    Phosphorylated on one or several of Thr-220, Ser-245, Ser-250, and Ser-255. In response to TGF-beta, phosphorylated on Ser-465/467 by TGF-beta and activin type 1 receptor kinases. TGF-beta-induced Ser-465/467 phosphorylation declines progressively in a KMT5A-dependent manner. Able to interact with SMURF2 when phosphorylated on Ser-465/467, recruiting other proteins, such as SNON, for degradation. In response to decorin, the naturally occurring inhibitor of TGF-beta signaling, phosphorylated on Ser-240 by CaMK2. Phosphorylated by MAPK3 upon EGF stimulation; which increases transcriptional activity and stability, and is blocked by calmodulin. Phosphorylated by PDPK1;In response to TGF-beta, ubiquitinated by NEDD4L; which promotes its degradation. Monoubiquitinated, leading to prevent DNA-binding (By similarity). Deubiquitination by USP15 alleviates inhibition and promotes activation of TGF-beta target genes (PubMed:21947082). Ubiquitinated by RNF111, leading to its degradation: only SMAD2 proteins that are 'in use' are targeted by RNF111, RNF111 playing a key role in activating SMAD2 and regulating its turnover (By similarity);Acetylated on Lys-19 by coactivators in response to TGF-beta signaling, which increases transcriptional activity. Isoform short: Acetylation increases DNA binding activity in vitro and enhances its association with target promoters in vivo. Acetylation in the nucleus by EP300 is enhanced by TGF-beta

  • Subunit

    Monomer; in the absence of TGF-beta (PubMed:9670020). Heterodimer; in the presence of TGF-beta (PubMed:9670020). Forms a heterodimer with co-SMAD, SMAD4, in the nucleus to form the transactivation complex SMAD2/SMAD4 (PubMed:15350224, PubMed:24324267, PubMed:9670020). Found in a complex with SMAD3 and TRIM33 upon addition of TGF-beta (PubMed:16751102). Identified in a complex that contains at least ZNF451, SMAD2, SMAD3 and SMAD4 (PubMed:24324267). Interacts (via the MH2 domain) with ZFYVE9; may form trimers with the SMAD4 co-SMAD (PubMed:10615055). Interacts with TAZ/WWRT1 (PubMed:18568018). Interacts with FOXH1 (PubMed:9702198). Interacts with SNW1 (PubMed:11278756). Interacts with CREB-binding protein (CBP) and EP300 (PubMed:16862174). Interacts with SNON (PubMed:11389444). Interacts with ALK4/ACVR1B (PubMed:10615055, PubMed:9892009). Interacts with SKOR1 (PubMed:17292623). Interacts with SKOR2 (PubMed:16200078). Interacts with PRDM16 (PubMed:19049980). Interacts (via MH2 domain) with LEMD3 (PubMed:15601644, PubMed:15647271). Interacts with RBPMS (PubMed:17099224). Interacts with WWP1. Interacts (dephosphorylated form, via the MH1 and MH2 domains) with RANBP3 (via its C-terminal R domain); the interaction results in the export of dephosphorylated SMAD3 out of the nucleus and termination of the TGF-beta signaling (PubMed:19289081). Interacts with PDPK1 (via PH domain) (PubMed:17327236). Interacts with DAB2; the interactions are enhanced upon TGF-beta stimulation (PubMed:11387212). Interacts with USP15 (PubMed:21947082). Interacts with PPP5C (PubMed:22781750). Interacts with LDLRAD4 (via the SMAD interaction motif) (PubMed:24627487). Interacts (via MH2 domain) with PMEPA1 (via the SMAD interaction motif) (PubMed:20129061). Interacts with ZFHX3 (PubMed:25105025). Interacts with ZNF451 (PubMed:24324267). Interacts with SMURF2 when phosphorylated on Ser-465/467 (PubMed:11389444). Interacts with PPM1A (PubMed:16751101). Interacts with TGF-beta (PubMed:8980228). Interacts with TGFBR1 (PubMed:9865696). Interacts with TGIF (PubMed:10835638). Interacts with SMAD3 and TRIM33 (PubMed:16751102). Interacts with ZNF580 (PubMed:21599657). Interacts with NEDD4L in response to TGF-beta (By similarity). Interacts with HGS (By similarity). Interacts with AIP1 (By similarity). Interacts with WWP1 (By similarity). Interacts with PML (By similarity). Interacts weakly with ZNF8 (By similarity). Interacts (when phosphorylated) with RNF111; RNF111 acts as an enhancer of the transcriptional responses by mediating ubiquitination and degradation of SMAD2 inhibitors (By similarity). Interacts with YAP1 (when phosphorylated at 'Ser-127') (By similarity). Interacts when phosphorylated with IPO7; the interaction facilitates translocation of SMAD2 to the nucleus (By similarity). Interacts with MTMR4; negatively regulates TGF-beta signaling through SMAD2 dephosphorylation and retention in endosomes (Probable)

  • SwissProt ID

    Q15796

  • Gene ID
    4087 [NCBI]
  • Synonyms

    SMAD2; MADH2; MADR2; Mothers against decapentaplegic homolog 2; MAD homolog 2; Mothers against DPP homolog 2; JV18-1; Mad-related protein 2; hMAD-2; SMAD family member 2; SMAD 2; Smad2; hSMAD2

  • Research Field

    Signal Transduction

[1]. Eppert K, et al. MADR2 maps to 18q21 and encodes a TGFbeta-regulated MAD-related protein that is functionally mutated in colorectal carcinoma. Cell. 1996 Aug 23;86(4):543-52. [Content Brief]

Smad2/3 Antibody Related Classifications

MOQ
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100 mg

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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(20ug) , HEK293(lane 3(20ug) and C6(lane 4(20ug) using Smad2/3 Antibody (HY-P80893) 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-P83730, 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
     Western blot analysis was performed on extracts from Hela (lane 1, 15 μg), HT-1080 (lane 2, 15 μg), A549 (lane 3, 15 μg), LnCaP (lane 4, 15 μg), RAW264.7 (lane 5, 15 μg), and C6 (lane 6, 15 μg) using Smad2/3 Rabbit pAb. Proteins were transferred to a PVDF membrane and blocked with 5% non - fat milk in TBST at 4°C overnight. The primary antibody (1:1000 dilution) and the loading control antibody (beta-Actin(HRP), HY-P80993, 1:5000 dilution) was incubated in 5% non-fat milk in TBST for 1 hour at 37°C. Goat Anti - Rabbit IgG - HRP Secondary Antibody (1:20000 dilution) was then applied for 40 minutes at 37°C.

  • 1/16

    MedChemExpress Validation
     Flow cytometric analysis of 1X106 NIH-3T3 cells labeling Smad2/3 Antibody(HY-P80893, red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Then stained with the primary antibody at 1/100 dilution for an hour at 4℃. AF488-conjugated Goat Anti-Rabbit IgG H&L (HY-P8002) was used as the secondary antibody at 1/1,000 dilution for 30 minutes at 4℃. Rabbit IgG Isotype Control (HY-P80879, blue) was used as the isotype control, cells without incubation with primary antibody were used as the unlabeled control (black).

  • 1/16

    MedchemExpress Validation
     Immunocytochemistry analysis of Hela cells labeling Smad2/3 Antibody (HY-P80893) 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 Smad2/3 Antibody (HY-P80893) 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 A549 cells labeling Smad2/3 Antibody (HY-P80893) 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 Smad2/3 Antibody (HY-P80893) 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).

  • Smad2/3 Antibody Product Image
    1/16
  • Experimental Validation Results for Smad2/3 Antibody
  • Experimental Validation Results for Smad2/3 Antibody
  • Experimental Validation Results for Smad2/3 Antibody
  • Experimental Validation Results for Smad2/3 Antibody
  • Experimental Validation Results for Smad2/3 Antibody
  • Smad2/3 Antibody Product Image
  • Experimental Validation Results for Smad2/3 Antibody
    1/1
    MedChemExpress Validation
     Western blot analysis of extracts from Hela(lane 2(20ug) , HEK293(lane 3(20ug) and C6(lane 4(20ug) using Smad2/3 Antibody (HY-P80893) 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-P83730, 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 Smad2/3 Antibody
    1/1
    MedChemExpress Validation
     Western blot analysis was performed on extracts from Hela (lane 1, 15 μg), HT-1080 (lane 2, 15 μg), A549 (lane 3, 15 μg), LnCaP (lane 4, 15 μg), RAW264.7 (lane 5, 15 μg), and C6 (lane 6, 15 μg) using Smad2/3 Rabbit pAb. Proteins were transferred to a PVDF membrane and blocked with 5% non - fat milk in TBST at 4°C overnight. The primary antibody (1:1000 dilution) and the loading control antibody (beta-Actin(HRP), HY-P80993, 1:5000 dilution) was incubated in 5% non-fat milk in TBST for 1 hour at 37°C. Goat Anti - Rabbit IgG - HRP Secondary Antibody (1:20000 dilution) was then applied for 40 minutes at 37°C.
  • Experimental Validation Results for Smad2/3 Antibody
    1/1
    MedChemExpress Validation
     Flow cytometric analysis of 1X106 NIH-3T3 cells labeling Smad2/3 Antibody(HY-P80893, red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Then stained with the primary antibody at 1/100 dilution for an hour at 4℃. AF488-conjugated Goat Anti-Rabbit IgG H&L (HY-P8002) was used as the secondary antibody at 1/1,000 dilution for 30 minutes at 4℃. Rabbit IgG Isotype Control (HY-P80879, blue) was used as the isotype control, cells without incubation with primary antibody were used as the unlabeled control (black).
  • Experimental Validation Results for Smad2/3 Antibody
    1/1
    MedchemExpress Validation
     Immunocytochemistry analysis of Hela cells labeling Smad2/3 Antibody (HY-P80893) 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 Smad2/3 Antibody (HY-P80893) 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 Smad2/3 Antibody
    1/1
    MedchemExpress Validation
     Immunocytochemistry analysis of A549 cells labeling Smad2/3 Antibody (HY-P80893) 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 Smad2/3 Antibody (HY-P80893) 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 Smad2/3 Antibody
  • Experimental Validation Results for Smad2/3 Antibody
  • Experimental Validation Results for Smad2/3 Antibody
  • Experimental Validation Results for Smad2/3 Antibody
  • Experimental Validation Results for Smad2/3 Antibody
  • Smad2/3 Antibody Product Image
    MedChemExpress Validation

    Western blot analysis of extracts from Hela(lane 2(20ug) , HEK293(lane 3(20ug) and C6(lane 4(20ug) using Smad2/3 Antibody (HY-P80893) 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-P83730, 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.

  • Smad2/3 Antibody Product Image
    MedChemExpress Validation

    Western blot analysis was performed on extracts from Hela (lane 1, 15 μg), HT-1080 (lane 2, 15 μg), A549 (lane 3, 15 μg), LnCaP (lane 4, 15 μg), RAW264.7 (lane 5, 15 μg), and C6 (lane 6, 15 μg) using Smad2/3 Rabbit pAb. Proteins were transferred to a PVDF membrane and blocked with 5% non - fat milk in TBST at 4°C overnight. The primary antibody (1:1000 dilution) and the loading control antibody (beta-Actin(HRP), HY-P80993, 1:5000 dilution) was incubated in 5% non-fat milk in TBST for 1 hour at 37°C. Goat Anti - Rabbit IgG - HRP Secondary Antibody (1:20000 dilution) was then applied for 40 minutes at 37°C.

  • Smad2/3 Antibody Product Image
    MedChemExpress Validation

    Flow cytometric analysis of 1X106 NIH-3T3 cells labeling Smad2/3 Antibody(HY-P80893, red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Then stained with the primary antibody at 1/100 dilution for an hour at 4℃. AF488-conjugated Goat Anti-Rabbit IgG H&L (HY-P8002) was used as the secondary antibody at 1/1,000 dilution for 30 minutes at 4℃. Rabbit IgG Isotype Control (HY-P80879, blue) was used as the isotype control, cells without incubation with primary antibody were used as the unlabeled control (black).

  • Smad2/3 Antibody Product Image
    MedchemExpress Validation

    Immunocytochemistry analysis of Hela cells labeling Smad2/3 Antibody (HY-P80893) 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 Smad2/3 Antibody (HY-P80893) 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).

  • Smad2/3 Antibody Product Image
    MedchemExpress Validation

    Immunocytochemistry analysis of A549 cells labeling Smad2/3 Antibody (HY-P80893) 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 Smad2/3 Antibody (HY-P80893) 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).

  • Smad2/3 Antibody Product Image WB
  • Smad2/3 Antibody Product Image WB
  • Smad2/3 Antibody Product Image FC
  • Smad2/3 Antibody Product Image ICC
  • Smad2/3 Antibody Product Image ICC