Anti-Smad4 antibody [EP618Y] - BSA and Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [EP618Y] - BSA and Azide free (AB215968)

This IHC data was generated using the same anti-Smad4 antibody clone, EP618Y, in a different buffer formulation (cat# ab40759).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue labelling Smad4 with purified ab40759 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [EP618Y] - BSA and Azide free (AB215968)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma tissue labelling Smad4 with unpurified ab40759 at a 1/100 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40759).

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• IHC-P

AbReview31424****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [EP618Y] - BSA and Azide free (AB215968)

Unpurified ab40759 staining Smad4 in rat femur tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% BSA for 20 minutes at 22°C; antigen retrieval was by heat mediation in a citrate buffer pH6.0. Samples were incubated with primary antibody (1/200 in blocking buffer) for 2 hours at 20°C. An undiluted HRP-conjugated goat anti-rabbit IgG polyclonal (1/250) was used as the secondary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40759).

This image is courtesy of an anonymous Abreview.

• IHC-P

AbReview46882****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [EP618Y] - BSA and Azide free (AB215968)

4% PFA-fixed, paraffin-embedded postnatal day 21 mouse ovary tissue stained for Smad4 using ab40759 at 1/500 dilution in immunohistochemical analysis.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40759).

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

This image is courtesy of an anonymous Abreview.

• WB

Lab

Western blot - Anti-Smad4 antibody [EP618Y] - BSA and Azide free (AB215968)

This data was developed using ab40759, the same antibody clone in a different buffer formulation. Different batches of ab40759 were tested on Ramos (Human Burkitt's lymphoma B lymphocyte) lysate at 0.7 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 60 kDa.

All lanes:

Western blot - Anti-Smad4 antibody [EP618Y] (<a href='/en-us/products/primary-antibodies/smad4-antibody-ep618y-ab40759'>ab40759</a>)

Predicted band size: 60 kDa

false

• WB

Lab

Western blot - Anti-Smad4 antibody [EP618Y] - BSA and Azide free (AB215968)

This WB data was generated using the same anti-Smad4 antibody clone, EP618Y, in a different buffer formulation (cat# ab40759).

Lane 1 : Wild type HAP1 whole cell lysate (20 μg)
Lane 2 : SMAD4 knockout HAP1 whole cell lysate (20 μg)
Lane 3 : HepG2 whole cell lysate (20 μg)
Lane 4 : Jurkat whole cell lysate (20 μg)

Lanes 1 - 4 : Merged signal (red and green). Green - ab40759 observed at 60 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab40759 was shown to specifically react with SMAD4 in wild type HAP1 cells. No band was observed when SMAD4 knockout HAP1 samples were used. Wild-type and SMAD4 knockout samples were subjected to SDS-PAGE. ab40759 and ab9484 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Smad4 antibody [EP618Y] (<a href='/en-us/products/primary-antibodies/smad4-antibody-ep618y-ab40759'>ab40759</a>)

Predicted band size: 60 kDa

false