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Smad3 (3D1) Monoclonal Antibody

Lane 1: A549 Cell lysates; probed with Smad3 (3D1) Monoclonal Antibody (bsm-52224R) at 1:1000 overnight at 4˚C. Followed by a conjugated secondary antibody.
Lane 1: A549 Cell lysates; probed with Smad3 (3D1) Monoclonal Antibody (bsm-52224R) at 1:1000 overnight at 4˚C. Followed by a conjugated secondary antibody.
SW480 cells were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100\/PBS and stained with Smad3 (3D1) Monoclonal Antibody (bsm-52224R) at 1:200 and incubated overnight at 4C, followed by secondary antibody incubation, DAPI staining of the nuclei and detection.
SW480 cells were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100\/PBS and stained with Smad3 (3D1) Monoclonal Antibody (bsm-52224R) at 1:200 and incubated overnight at 4C, followed by secondary antibody incubation, DAPI staining of the nuclei and detection.
Paraformaldehyde-fixed, paraffin embedded human lung cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 30 minutes; Blocking buffer at 37\u00b0C for 20min; Antibody incubation with Smad3 (3D1) Monoclonal Antibody (bsm-52224R) at 1:50 overnight at 4\u00b0C, followed by a conjugated secondary and DAB staining.
Paraformaldehyde-fixed, paraffin embedded human lung cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 30 minutes; Blocking buffer at 37\u00b0C for 20min; Antibody incubation with Smad3 (3D1) Monoclonal Antibody (bsm-52224R) at 1:50 overnight at 4\u00b0C, followed by a conjugated secondary and DAB staining.
Paraformaldehyde-fixed, paraffin embedded human lung cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 30 minutes; Blocking buffer at 37\u00b0C for 20min; Antibody incubation with Smad3 (3D1) Monoclonal Antibody (bsm-52224R) at 1:50 overnight at 4\u00b0C, followed by a conjugated secondary and DAB staining.
Paraformaldehyde-fixed, paraffin embedded human lung cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 30 minutes; Blocking buffer at 37\u00b0C for 20min; Antibody incubation with Smad3 (3D1) Monoclonal Antibody (bsm-52224R) at 1:50 overnight at 4\u00b0C, followed by a conjugated secondary and DAB staining.
Flow cytometric analysis of Hela cells with Smad3 (3D1) Monoclonal Antibody (bsm-52224R) at 1\/50 dilution (red) compared with an unlabelled control (cells without incubation with primary antibody; black). Alexa Fluor 488-conjugated goat anti rabbit IgG was used as the secondary antibody.
Flow cytometric analysis of Hela cells with Smad3 (3D1) Monoclonal Antibody (bsm-52224R) at 1\/50 dilution (red) compared with an unlabelled control (cells without incubation with primary antibody; black). Alexa Fluor 488-conjugated goat anti rabbit IgG was used as the secondary antibody.
Lane 1: Mouse Cerebrum lysates; Lane 2: Rat Cerebrum lysates; Lane 3: Mouse Large intestine lysates; Lane 4: Mouse Ovary lysates ; Lane 5: Mouse Lung lysates probed with Smad3 Monoclonal Antibody, Unconjugated (bsm-52224R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
Lane 1: Mouse Cerebrum lysates; Lane 2: Rat Cerebrum lysates; Lane 3: Mouse Large intestine lysates; Lane 4: Mouse Ovary lysates ; Lane 5: Mouse Lung lysates probed with Smad3 Monoclonal Antibody, Unconjugated (bsm-52224R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
Paraformaldehyde-fixed, paraffin embedded Mouse liver; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with Smad3 (3D1) Monoclonal Antibody, Unconjugated (bsm-52224R) at 1:200 overnight at 4\u00b0C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Mouse liver; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with Smad3 (3D1) Monoclonal Antibody, Unconjugated (bsm-52224R) at 1:200 overnight at 4\u00b0C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Rat brain; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with Smad3 (3D1) Monoclonal Antibody, Unconjugated (bsm-52224R) at 1:200 overnight at 4\u00b0C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Rat brain; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with Smad3 (3D1) Monoclonal Antibody, Unconjugated (bsm-52224R) at 1:200 overnight at 4\u00b0C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Mouse ovarian; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with Smad3 (3D1) Monoclonal Antibody, Unconjugated (bsm-52224R) at 1:200 overnight at 4\u00b0C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Mouse ovarian; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with Smad3 (3D1) Monoclonal Antibody, Unconjugated (bsm-52224R) at 1:200 overnight at 4\u00b0C, DAB staining.
HUVEC cells were fixed with 4% PFA for 10min at room temperature,permeabilized with 0.1% PBST for 20 min at room temperature, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with Smad3 (3D1) Monoclonal Antibody(bsm-52224R)at 1:100 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).
HUVEC cells were fixed with 4% PFA for 10min at room temperature,permeabilized with 0.1% PBST for 20 min at room temperature, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with Smad3 (3D1) Monoclonal Antibody(bsm-52224R)at 1:100 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).
Raw264.7 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37\u00b0C for 20 min; Antibody incubation with (Smad3) monoclonal Antibody, Unconjugated (bsm-52224R) 1:100, 90 minutes at 37\u00b0C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37\u00b0C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.
Raw264.7 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37\u00b0C for 20 min; Antibody incubation with (Smad3) monoclonal Antibody, Unconjugated (bsm-52224R) 1:100, 90 minutes at 37\u00b0C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37\u00b0C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.

Applications

  • WB
  • FCM
  • IHC-P
  • IF(ICC)

Reactivity

  • Human
  • Mouse
  • Rat
Overview
Catalog # bsm-52224R
Product Name Smad3 (3D1) Monoclonal Antibody
Applications WB, FCM, IHC-P, IF(ICC)
Reactivity Human, Mouse, Rat
Specifications
Conjugation Unconjugated
Host Rabbit
Source Recombinant human Smad3 protein, around N-terminal 1-200aa.
Clonality Monoclonal
Clone # 3D1
Isotype IgG
Concentration 1ug/ul
Purification Purified by Protein A.
Storage Buffer Aqueous buffered solution containing 1xTBS (pH7.4), 1%BSA, 40%Glycerol and 0.05% Sodium Azide.
Storage Condition Shipped at 4°C. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.
Target
Gene ID 4088
Swiss Prot P84022
Synonyms Smad3 phospho Ser423; Smad3 phospho S423; p-Smad3 Ser423; hMAD 3; hSMAD3; HSPC193; JV15 2; JV152; MAD mothers against decapentaplegic Drosophila homolog 3; MAD3; MADH 3; MADH3; Mothers against decapentaplegic homolog 3; Mothers against DPP homolog 3; SMA and MAD related protein 3; SMAD 3; SMAD; SMAD-3; SMAD3_HUMAN.
Background Smad3 is a 50 kDa member of a family of proteins that act as key mediators of TGF beta superfamily signaling in cell proliferation, differentiation and development. The Smad family is divided into three subclasses: receptor regulated Smads, activin/TGF beta receptor regulated (Smad2 and 3) or BMP receptor regulated (Smad 1, 5, and 8); the common partner, (Smad4) that functions via its interaction to the various Smads; and the inhibitory Smads, (Smad6 and 7). Activated Smad3 oligomerizes with Smad4 upon TGF beta stimulation and translocates as a complex into the nucleus, allowing its binding to DNA and transcription factors. Phosphorylation of the two TGF beta dependent serines 423 and 425 in the C terminus of Smad3 is critical for Smad3 transcriptional activity and TGF beta signaling.
Application Dilution
WB 1:300-5000
FCM 1:20-100
IHC-P 1:200-400
IF(ICC) 1:50-200

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