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. (Ship on dry ice.)

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.

Cross-reactivity

No cross reactivity with other proteins.

Reactive Species

P00059 is reactive to SMAD3 in Human

Calculated molecular weight

48.1 kDa

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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Applications

P00059 is guaranteed for ELISA, Flow Cytometry, IHC, WB Boster Guarantee

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
This affinity purified antibody has been tested for use in ELISA, immunohistochemistry, and western blot.  Specific conditions for reactivity should be optimized by the end user. Expect a band approximately 48 kDa in size corresponding to phosphorylated Smad3 protein by western blotting in the appropriate stimulated tissue or cell lysate or extract.  Less than 0.2% reactivity is observed against the non-phosphorylated form of the immunizing peptide.  This antibody is phospho specific for dual phosphorylated pS423 and pS425 of Smad3. Stimulation with 2 ng/ml TGF-beta for 1 hour is suggested.

Validation Images & Assay Conditions

See full target information SMAD3

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