Anti-Bcl-2 antibody [EPR17509] - BSA and Azide free

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Bcl-2 antibody [EPR17509] - BSA and Azide free (AB219608)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling Bcl-2 with ab182858 at 1/250 (red) compared with a rabbit monoclonal IgG isotype control (ab172730) (black) and a unlabelled control (cells without incubation with primary antibody and secondary antibody (blue)). Goat anti rabbit IgG (FITC) at 1/500 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 (ab182858).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [EPR17509] - BSA and Azide free (AB219608)

Immunohistochemical analysis of paraffin-embedded Human endometrial cancer tissue labeling Bcl-2 with ab182858 at 1/1000 followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500.

Cytoplasm, nuclear membrane and nucleus staining on lymphocytes and cancer cells of Human endometrial cancer tissue is observed.

Counter stained with Hematoxylin.
Negative control : Used PBS instead of primary antibody followed by ab97051 at 1/500.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab182858).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [EPR17509] - BSA and Azide free (AB219608)

Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling Bcl-2 with ab182858 at 1/1000 followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500.

Cytoplasm, nuclear membrane and nucleus staining on lymphocytes of Human tonsil tissue is observed.

Counter stained with Hematoxylin.

Negative control : Used PBS instead of primary antibody followed by ab97051 at 1/500.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab182858).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [EPR17509] - BSA and Azide free (AB219608)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling Bcl-2 with ab182858 at 1/1000 followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500.

Cytoplasm, nuclear membrane and nucleus staining on lymphocytes of Mouse spleen tissue is observed.

Counter stained with Hematoxylin.
Negative control : Used PBS instead of primary antibody followed by ab97051 at 1/500.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab182858).

• WB

Lab

Western blot - Anti-Bcl-2 antibody [EPR17509] - BSA and Azide free (AB219608)

This data was developed using the same antibody clone in a different buffer formulation (ab182858).

Lanes 1- 2 : Merged signal (red and green). Green - ab182858 observed at 26 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab182858 was shown to react with Bcl-2 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab255364 (knockout cell lysate ab263752) was used. Wild-type HeLa and BCL2 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab182858 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 2000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Bcl-2 antibody [EPR17509] (<a href='/en-us/products/primary-antibodies/bcl-2-antibody-epr17509-ab182858'>ab182858</a>) at 1/2000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

BCL2 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human BCL2 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-bcl2-knockout-hela-cell-line-ab255364'>ab255364</a>)

Predicted band size: 26 kDa

Observed band size: 26 kDa

false

• WB

Lab

Western blot - Anti-Bcl-2 antibody [EPR17509] - BSA and Azide free (AB219608)

This data was developed using the same antibody clone in a different buffer formulation (ab182858).

Lanes 1 - 4 : Merged signal (red and green). Green - ab182858 observed at 26 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab182858 was shown to specifically react with BCL2 when BCL2 knockout samples were used. Wild-type and BCL2 knockout samples were subjected to SDS-PAGE. ab182858 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1 ug/ml and 1/10000 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 1/10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Bcl-2 antibody [EPR17509] - BSA and Azide free (ab219608) at 1 µg/mL

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg/mL

Lane 2:

BCL2 knockout HAP1 whole cell lysate at 20 µg/mL

Lane 3:

HeLa whole cell lysate at 20 µg/mL

Lane 4:

THP-1 whole cell lysate at 20 µg/mL

Predicted band size: 26 kDa

Observed band size: 26 kDa

false