Smad2/3 [p Ser423, p Ser425] Antibody [Unconjugated]

Western blot shows lysates of Mv1Lu mink lung epithelial cell line untreated (-) or treated (+) with 10 ng/mL Recombinant Human TGF-beta 2 (Catalog # 302-B2) for 24 hours. PVDF membrane was probed with 1:500 dilution of Rabbit Anti-Human Phospho-Smad2 (S465/S467)/ Smad3 (S423/S425)Polyclonal Antibody (Catalog # AB3226), followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). Specific bands were detected for Phospho-Smad2 (S465/S467)/ Smad3 (S423/S425) at approximately 60 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Simple Western lane view shows lysates of HEK293T human embryonic kidney cell line untreated (-) or treated (+) with 200 nM PMA for 20 minutes, loaded at 0.5 mg/mL. A specific band was detected for Phospho-Smad2 (S465/S467)/Smad3 (S423/S425) at approximately 68 kDa (as indicated) using 1:50-1:100 dilution of Rabbit Anti-Human Phospho-Smad2 (S465/S467)/Smad3 (S423/S425) Polyclonal Antibody (Catalog # AB3226). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Smad2/3 phosphorylated at S465/S467 and S423/S425 was detected in immersion fixed Mv1Lu mink lung epithelial cell line treated with 10 ng/mL of Recombinant Human TGF-beta 2 (Catalog # 302-B2) using Rabbit Anti-Human Phospho-Smad2 (S465/S467)/ Smad3 (S423/S425) Polyclonal Antibody (Catalog # AB3226) at 1:50-1:100 dilution for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

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Smad2/3 [p Ser423, p Ser425] Antibody [Unconjugated] Summary

• Immunogen: Phosphopeptide containing human Smad3 S423/S425 sites
• Modification: p Ser423, p Ser425
• Specificity: Detects human Phospho-Smad2/3 when dually phosphorylated at S465/S467 and S423/S425, respectively.
• Source: N/A
• Isotype: Serum
• Clonality: Polyclonal
• Host: Rabbit
• Gene: SMAD3
• Purity Statement: N/A

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 1:50-1:100 dilution
  • Simple Western 1:50-1:100 dilution
  • Western Blot 1:500 dilution
• Application Notes: In Simple Western only 10-15 uL of the recommended dilution is used per data point.

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from rabbit serum in PBS and Sodium Azide.
• Preservative: Sodium Azide
• Reconstitution Instructions: Reconstitute with 250 μL sterile PBS containing 0.02% Sodium Azide.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Smad2/3 [p Ser423, p Ser425] Antibody [Unconjugated]

• hMAD-2
• hSMAD2
• JV18
• JV18-1SMAD, mothers against DPP homolog 2 (Drosophila)
• MAD homolog 2
• Mad protein homolog
• Mad, mothers against decapentaplegic homolog 2
• MADH2MGC34440
• MADR2MGC22139
• Mad-related protein 2
• mother against DPP homolog 2
• mothers against decapentaplegic homolog 2
• Mothers against DPP homolog 2
• Sma- and Mad-related protein 2
• SMAD 2
• SMAD family member 2MAD, mothers against decapentaplegic homolog 2 (Drosophila)
• SMAD, mothers against DPP homolog 2
• Smad2
• Smad2/3

Background

Smad3 (mothers against DPP homolog 3) is a transcription factor that functions downstream of the transforming growth factor beta receptor (TGF-beta R). Smad3 is phosphorylated in cells stimulated with TGF-beta , complexes with Smad4, and translocates to the nucleus to upregulate gene transcription. Smad3 is critical for signaling fibrosis and wound healing.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Smad2/3 Antibody (AB3226)(3)

Publications using AB3226

• Leung L, Radulovich N, Zhu C, Wang D, To C, Ibrahimov E, Tsao M Loss of canonical Smad4 signaling promotes KRAS driven malignant transformation of human pancreatic duct epithelial cells and metastasis. PLoS ONE, 2013-12-27;8(12):e84366. 2013-12-27 [PMID: 24386371] (Western Blot, Human)
• Yeh YH, Kuo CT, Chan TH, Chang GJ, Qi XY, Tsai F, Nattel S, Chen WJ Transforming growth factor-beta and oxidative stress mediate tachycardia-induced cellular remodelling in cultured atrial-derived myocytes. Cardiovasc. Res., 2011-02-02;91(1):62-70. 2011-02-02 [PMID: 21289011] (ICC, Western Blot, Mouse)
• Xing D, Bonanno JA Effect of cAMP on TGFbeta1-induced corneal keratocyte-myofibroblast transformation. Invest. Ophthalmol. Vis. Sci., 2008-10-20;50(2):626-33. 2008-10-20 [PMID: 18936144] (Western Blot, Rabbit)

Review for Smad2/3 Antibody (AB3226) (1)

Average Rating: 5

(Based on 1 review)

We have 1 review tested in 1 species: Mouse.

reviewed by: Verified Customer WB Mouse 01/05/2015

Summary

• Application: Western Blot
• Sample Tested: See PMID 22868230
• Species: Mouse

Bioinformatics

• Gene Symbol: SMAD3