Anti-Smad3 antibody [EP568Y]

• Supplier: Abcam
• Catalog: ab40854
• Prices: $404.00
• Sizes: 100 µl
• Host: Rabbit
• Clonality: Monoclonal
• Isotype: IgG
• Clone: EP568Y
• Applications: WB IHC-P ELISA FC ICC/IF ICC/IF
• Species Reactivities: Mouse, Rat, Human
• Antigen: Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Smad3 aa 200-300 (internal sequence)
• Blocking Peptide: Smad3 peptide
• Description: Rabbit Monoclonal
• Gene: SMAD3
• Conjugate: Unconjugated

Product images

Anti-Smad3 antibody [EP568Y] (ab40854) at 1/1300 dilution (unpurified) + Rat liver tissue lysate at 10 µgSecondaryPeroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Anti-Smad3 antibody [EP568Y] (ab40854) at 1/5300 dilution (purified) + Rat liver tissue lysate at 10 µgSecondaryPeroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

All lanes : Anti-Smad3 antibody [EP568Y] (ab40854) at 1/1300 dilution (unpurified)Lane 1 : HT-29 cell lysateLane 2 : HT-1080 cell lysateLysates/proteins at 10 µg per lane.SecondaryPeroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

All lanes : Anti-Smad3 antibody [EP568Y] (ab40854) at 1/5300 dilution (purified)Lane 1 : HT-29 cell lysateLane 2 : HT-1080 cell lysateLysates/proteins at 10 µg per lane.SecondaryPeroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Anti-Smad3 antibody [EP568Y] (ab40854) at 1/5000 dilution (unpurified) + Jurkat cell lysate at 10 µg

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue labelling Smad3 with unpurified ab40854 at 1/500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Counterstained with Hematoxylin.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue labelling Smad3 with purified ab40854 at 1/2000. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Counterstained with Hematoxylin.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostate carcinoma tissue labelling unpurified ab40854 at 1/100 dilution.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue labelling unpurified ab40854.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colonic adenocarcinoma tissue labelling unpurified ab40854.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung adenocarcinoma tissue labelling unpurified ab40854.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human glioma tissue labelling unpurified ab40854.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human gastric adenocarcinoma tissue labelling unpurified ab40854.

Immunocytochemsitry/Immunofluorescence analysis of HepG2 cells labelling Smad3 (green) with unpurified ab40854 at 1/500. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/200) was used as the secondary antibody. Counterstained with DAPI (blue).

Immunocytochemsitry/Immunofluorescence analysis of HepG2 cells labelling Smad3 (green) with purified ab40854 at 1/2000. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/200) was used as the secondary antibody. Counterstained with DAPI (blue).

Overlay histogram showing HCT116 cells stained with unpurified ab40854 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab40854, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

Flow cytometry analysis of HT-29 cells labelling Smad3 with unpurified ab40854 at 1/50 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Green - Isotype control, rabbit monoclonal IgG.

Flow cytometry analysis of HT-29 cells labelling Smad3 with purified ab40854 at 1/210 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Green - Isotype control, rabbit monoclonal IgG.

Standard Curve for Smad3 (Analyte: Smad3 protein (His tag) (ab89353, unpurified)); dilution range 1pg/ml to 1µg/ml using Capture Antibody Mouse monoclonal [AF9F7] to Smad3 (ab75512) at 5µg/ml and Detector Antibody Rabbit monoclonal [EP568Y] to Smad3 (ab40854) at 0.5µg/ml.

ab40854 staining Smad3 in human granulosa cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with ethanol and triton and blocked for 1 hour at 26°C. Samples were incubated with primary antibody (1/200) for 16 hours at 4°C. An undiluted IRDye® 800CW-conjugated goat anti-rabbit IgG (H+L) polyclonal was used as the secondary antibody. Left - negative control (4 replicates).See Abreview

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