Smad3 [p Ser423, p Ser425] Antibody (ST0493)

Novus Biologicals | Catalog # NBP2-67513

Recombinant Monoclonal Antibody
Novus Biologicals
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Key Product Details

Validated by

Biological Validation

Species Reactivity

Human, Mouse

Applications

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Immunocytochemistry/ Immunofluorescence

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Clone # ST0493 expressed in HEK293
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Product Specifications

Immunogen

Synthetic phospho-peptide corresponding to residues surrounding Ser423 and 425 of human Smad3. (SwissProt: P84022 Human; SwissProt: Q8BUN5 Mouse)

Modification

p Ser423, p Ser425

Localization

Cytoplasm, Nucleus.

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for Smad3 [p Ser423, p Ser425] Antibody (ST0493)

Western Blot: Smad3 [p Ser423, p Ser425] Antibody (ST0493) [NBP2-67513] - Analysis of Smad3 [p Ser423, p Ser425] on A549 cell lysates. Lane 1 : A549 cells, whole cell lysate, 10 ug /lane. Lane 2 : A549 cells were treated with 5 ng/ml TGF beta1 for 24 hours, whole cell lysates, 10 ug/lane. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody Anti-Smad3 [p Ser423, p Ser425] (1/500), Anti-Smad3 antibody (1/500) and Anti-GAPDH antibody (1/10,000)was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG H&L (HRP) Secondary Antibody at 1:200,000 dilution was used for 1 hour at room temperature. Predicted band size: 48 kDa Observed band size: 60 kDa Exposure time: 3 seconds
Immunocytochemistry/Immunofluorescence: Smad3 [p Ser423, p Ser425] Antibody (ST0493) [NBP2-67513] - Staining Phospho-Smad3(S423/S425) in A431 cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
Immunohistochemistry-Paraffin: Smad3 [p Ser423, p Ser425] Antibody (ST0493) [NBP2-67513] - Analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-Phospho-Smad3 [p Ser423, p Ser425) antibody washed with ddH2O and PBS, and then probed with the primary antibody at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Western Blot: Smad3 [p Ser423, p Ser425] Antibody (ST0493) [NBP2-67513] - Analysis of Phospho-Smad3(S423/S425) on K562 cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody ( 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:5,000 dilution was used for 1 hour at room temperature.
Western Blot: Smad3 [p Ser423, p Ser425] Antibody (ST0493) [NBP2-67513] - Lane 1: K562 cells, whole cell lysate, 10ug/lane, Lane 2: K562 cells treated with 2.8ug/ul lambda-PP for 30 minutes, whole cell lysates, 10ug/lane. All lanes : Anti-Phospho-Smad3(S423/S425) antibody at 1:500 dilution. Anti-GAPDH antibody at 1:10,000 dilution. Goat Anti-Rabbit IgG H&L (HRP) (HA1001) at 1/200,000 dilution. Predicted band size: 48 kDa Observed band size: 55 kDa Blocking and diluting buffer: 5% BSA. Exposure time: 3 minutes 43 seconds
Western Blot: Smad3 [p Ser423, p Ser425] Antibody (ST0493) [NBP2-67513] - Analysis of Smad3 [p Ser423, p Ser425] on different lysates with Rabbit anti-Smad3 [p Ser423, p Ser425] antibody at 1/500 dilution. Lane 1: A549 cell lysate Lane 2: K562 cell lysate Lysates/proteins at 10 ug/Lane. Predicted band size: 48 kDa Observed band size: 60 kDa Exposure time: 2 minutes; 10% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:300,000 dilution was used for 1 hour at room temperature.
Immunohistochemistry-Paraffin: Smad3 [p Ser423, p Ser425] Antibody (ST0493) [NBP2-67513] - Analysis of paraffin-embedded mouse skin tissue using anti-Phospho-Smad3(S423/S425) antibody. Counter stained with hematoxylin.
Immunohistochemistry-Paraffin: Smad3 [p Ser423, p Ser425] Antibody (ST0493) [NBP2-67513] - Analysis of paraffin-embedded human large intestine tissue with Rabbit anti-Phospho-Smad3 [p Ser423, p Ser425] antibody washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemistry-Paraffin: Smad3 [p Ser423, p Ser425] Antibody (ST0493) [NBP2-67513] - Analysis of paraffin-embedded mouse skin tissue with Rabbit anti-Smad3 [p Ser423, p Ser425] antibody washed with ddH2O and PBS, and then probed with the primary antibody at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Applications for Smad3 [p Ser423, p Ser425] Antibody (ST0493)

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:50-1:200

Immunohistochemistry-Paraffin

1:50-1:500

Western Blot

1:500-1:2000

Formulation, Preparation, and Storage

Purification

Protein A purified

Formulation

TBS (pH7.4), 0.05% BSA, 40% Glycerol

Preservative

0.05% Sodium Azide

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: Smad3

The protein encoded by the Smad3 gene belongs to the SMAD, a family of proteins similar to the gene products of the Drosophila gene 'mothers against decapentaplegic' (Mad) and the C. elegans gene Sma. SMAD proteins are signal transducers and transcriptional modulators that mediate multiple signaling pathways. This protein functions as a transcriptional modulator activated by transforming growth factor-beta and is thought to play a role in the regulation of carcinogenesis. (provided by RefSeq)

Long Name

Mothers Against DPP Homolog 3

Alternate Names

DKFZp586N0721, DKFZp686J10186, hMAD-3, hSMAD3, HsT17436, JV15-2MGC60396, MAD homolog 3, mad protein homolog, MAD, mothers against decapentaplegic homolog 3, MAD, mothers against decapentaplegic homolog 3 (Drosophila), 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, SMAD, mothers against DPP homolog 3 (Drosophila), Smad3

Gene Symbol

SMAD3

Additional Smad3 Products

Product Documents for Smad3 [p Ser423, p Ser425] Antibody (ST0493)

Certificate of Analysis

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Product Specific Notices for Smad3 [p Ser423, p Ser425] Antibody (ST0493)

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.

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Protocols

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FAQs for Smad3 [p Ser423, p Ser425] Antibody (ST0493)

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  • Q: I was wondering if you can recommend any primary polyclonal Smad3 antibody (reactivity: human) for gel supershifts?

    A: We do provide a wide range of Smad3 antibodies, but unfortunately none of them have been tested for gel shift assay, so we can't guarantee they will work for an application never tested before.

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