bsm-52224R
[Primary Antibody]
Smad3 (3D1) Monoclonal Antibody
DATASHEET
Host: Rabbit
Target Protein: Smad3
Clonality: Monoclonal
Isotype: IgG
Entrez Gene: 4088
Swiss Prot: P84022
Source: Recombinant human Smad3 protein, around N-terminal 1-200aa.
Purification: Purified by Protein A.
Storage Buffer: 0.01M TBS(pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
Storage: Shipped at 4C. Store at -20C for one year. Avoid repeated freeze/thaw cycles.
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.
Size: 100ul
Concentration: 1ug/ul
Cross Reactive Species:
Human
Mouse
Rat
For research use only. Not intended for diagnostic or therapeutic use.
PRODUCT SPECIFIC PUBLICATIONS
- Li L et al. Anti-fibrotic effect of melittin on TRIM47 expression in Human embryonic lung fibroblast through regulating TRIM47 pathway. Life Sci. 2020 Sep 1;256:117893.Read more>>
- Huajun Wang. et al. LncRNA NEAT1 promotes proliferation, migration, invasion and epithelial-mesenchymal transition process in TGF-2-stimulated lens epithelial cells through regulating the miR-486-5p/SMAD4 axis. Cancer Cell Int. 2020 Dec;20(1):1-12Read more>>
- Wu Li-Juan. et al. Dahuang Zhechong Pills Suppress Silicosis Fibrosis Progression via p38 MAPK/TGF-1/Smad Pathway In Vitro. Evid-Based Compl Alt. 2021;2021:6662261Read more>>
VALIDATION IMAGES
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.
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°C for 20min; Antibody incubation with Smad3 (3D1) Monoclonal Antibody (bsm-52224R) at 1:50 overnight at 4°C, 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°C for 20min; Antibody incubation with Smad3 (3D1) Monoclonal Antibody (bsm-52224R) at 1:50 overnight at 4°C, 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.
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°C for 30min; Antibody incubation with Smad3 (3D1) Monoclonal Antibody, Unconjugated (bsm-52224R) at 1:200 overnight at 4°C, 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°C for 30min; Antibody incubation with Smad3 (3D1) Monoclonal Antibody, Unconjugated (bsm-52224R) at 1:200 overnight at 4°C, 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°C for 30min; Antibody incubation with Smad3 (3D1) Monoclonal Antibody, Unconjugated (bsm-52224R) at 1:200 overnight at 4°C, 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).
Raw264.7 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37°C for 20 min; Antibody incubation with (Smad3) monoclonal Antibody, Unconjugated (bsm-52224R) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.