Anti-Phospho-SMAD3 S423/S425 Antibody

Product Name

Anti-Phospho-SMAD3 S423/S425 Antibody

View all SMAD3 Antibodies

SKU/Catalog Number

P00059

Size

100ug

Form

Liquid (sterile filtered)

Description

Boster Bio Anti-Phospho-SMAD3 S423/S425 Antibody (Catalog # P00059). Tested in ELISA, IHC, WB applications. This antibody reacts with Human.

Storage & Handling

Store vial at -20°C prior to opening. Aliquot contents and freeze at -20°C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4°C as an undiluted liquid. Dilute only prior to immediate use. Expiration date is one (1) year from date of opening.

Cite This Product

Anti-Phospho-SMAD3 S423/S425 Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # P00059)

Host

Rabbit

Contents

0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2, 0.01% (w/v) Sodium Azide

Clonality

Polyclonal

Isotype

IgG

Immunogen

Anti-SMAD3 pS423pS425 antibody was prepared from whole rabbit serum produced by repeated immunizations with a dual phosphorylated synthetic peptide corresponding to a c-terminal region with Serine 423 and Serine 425 of human SMAD3 protein.

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross reactivity with other proteins.

Reactive Species

P00059 is reactive to SMAD3 in Human

Applications

P00059 is guaranteed for ELISA, IHC, WB Boster Guarantee

Observed Molecular Weight

Calculated molecular weight

48.081kDa

Background of SMAD3

This antibody is suitable for Cancer, Immunology and Nuclear Signaling research. Smad3 (also known as Mothers against decapentaplegic homolog 3 Mothers against DPP homolog 3, Mad3, hMAD-3, JV15-2 or hSMAD3) is a transcriptional modulator activated by TGF-beta (transforming growth factor) and activin type 1 receptor kinase. These activators exert diverse effects on a wide array of cellular processes. The Smad proteins mediate much of the signaling responses induced by the TGF-b superfamily. Briefly, activated type I receptor phosphorylates receptor-activated Smads (R-Smads) at their c-terminal two extreme serines in the SSXS motif, e.g. Smad2 and Smad3 proteins in the TGF-b pathway, or Smad1, Smad5 or Smad8 in the BMP pathway. Then the phosphorylated R-Smad translocated into nucleus, where they regulate transcription of target genes. Based on microarray and animal model experiments, Smad3 accounts for at least 80% of all TGF-b-mediated response.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

ELISA: 1:15,000 - 1:30,000
Flow Cytometry: User optimized
IHC: 1:500
WB: 1:2,000 - 1:20,000

Validation Images & Assay Conditions

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SMAD3 was detected in paraffin-embedded sections of epidermal keratinocytes at 40X using rabbit anti-SMAD3 phospho S423/phospho S425 Antigen Affinity purified polyclonal antibody (Catalog # P00059) at 2.5 μg/mL. The image shows localization of the antibody as the precipitated red signal, with a hematoxylin purple nuclear counterstain. Personal Communi-cation, Tina Roush, LifeSpanBiosciences, Seattle, WA.

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Western blot analysis of endogenous SMAD3 expression in stimulated cell lysates. Lysates were prepared from control cells (-lanes), or cells stimulated with 2 ng/ml TGF for 1 hour. Phosphorylated SMAD3 was detected using rabbit anti-SMAD3 phospho S423/phospho S425 Antigen Affinity purified polyclonal antibody (Catalog # P00059). Personal Communication. Ying Zhang, NIH, CCR, Bethesda, MD.

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Western blot analysis of SMAD3 expression in HEK293 lysate (lane 1), HeLa Lysate (lane 2), MCF-7 Lysate (lane 3), Jurkat Lysate (lane 4), A431 Lysate (lane 5), A549 Lysate (lane 6), LNCap Lysate (lane 7), MOLT-4 Lysate (lane 8), Ramos Lysate (lane 9), Raji Lsyate (lane 10), A-172 Lysate (lane 11) and NIH/3T3 Lysate (lane 12). Phosphorylated SMAD3 at 35KD was detected using rabbit anti-SMAD3 phospho S423/phospho S425 Antigen Affinity purified polyclonal antibody (Catalog # P00059) at 1ug/mL. The blot was developed using chemiluminescence (ECL) method (Catalog # EK1002).

Gene/Protein Information For SMAD3 (Source: Uniprot.org, NCBI)

Alternative Names

DKFZp586N0721; DKFZp686J10186; hMAD-3; hSMAD3; HsT17436; JV15-2MGC60396; MAD homolog 3; mad protein homolog; MAD, mothers against decapentaplegic homolog 3 (Drosophila); MAD, mothers against decapentaplegic homolog 3; mad3; MADH3mad homolog JV15-2; mothers against decapentaplegic homolog 3; Mothers against DPP homolog 3; SMA- and MAD-related protein 3; SMAD 3; SMAD family member 3HSPC193; SMAD, mothers against DPP homolog 3 (Drosophila); SMAD, mothers against DPP homolog 3; Smad3 SMAD3 HSPC193, HsT17436, JV15-2, LDS1C, LDS3, MADH3 SMAD family member 3 mothers against decapentaplegic homolog 3|MAD homolog 3|MAD, mothers against decapentaplegic homolog 3|SMA- and MAD-related protein 3|SMAD, mothers against DPP homolog 3|hMAD-3|hSMAD3|mad homolog JV15-2|mad protein homolog|mad3|mothers against DPP homolog 3

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