Anti-CD80 antibody [EPR1157(2)] - BSA and Azide free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD80 antibody [EPR1157(2)] - BSA and Azide free (AB271905)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling CD80 with ab134120 at a dilution of 1/1000. Heat mediated antigen retrieval was performed using Trs/EDTA buffer, pH9 (ab93684). ImmunoHistoProbe one step HRP Polymer was used. Counter stained with hematoxylin.

The image shows cytoplasmic and membrane staining.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134120).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-CD80 antibody [EPR1157(2)] - BSA and Azide free (AB271905)

This data was developed using the same antibody clone in a different buffer formulation containing PBS BSA glycerol and sodium azide (ab134120).

• IP

Lab

Immunoprecipitation - Anti-CD80 antibody [EPR1157(2)] - BSA and Azide free (AB271905)

This data was developed using the same antibody clone in a different buffer formulation (ab134120).

CD80 was immunoprecipitated from 0.35 mg Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate with ab134120 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab134120 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Lane 1:

Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate 10 μg

Lane 2:

Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of <a href='/en-us/products/primary-antibodies/cd80-antibody-epr11572-ab134120'>ab134120</a> in Raji whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>)

Observed band size: 60 kDa

false

Exposure time: 6s

• WB

Lab

Western blot - Anti-CD80 antibody [EPR1157(2)] - BSA and Azide free (AB271905)

This data was developed using the same antibody clone in a different buffer formulation (ab134120).

Lanes 1 - 3 : Merged signal (red and green). Green - ab134120 observed at 55 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.

ab134120 was shown to react with CD80 in wild-type Raji cells in western blot with loss of signal observed in CD80 knockout sample. Wild-type and CD80 knockout Raji cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween^®) before incubation with ab134120 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4°C at a 1 in 1000 Dilution and a 1 in 20000 dilution respectively. Blots were incubated 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 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-CD80 antibody [EPR1157(2)] (<a href='/en-us/products/primary-antibodies/cd80-antibody-epr11572-ab134120'>ab134120</a>) at 1/1000 dilution

Lane 1:

Wild-type Raji (Human Burkitt's lymphoma cell line) whole cell lysate at 20 µg

Lane 2:

CD80 knockout Raji (Human Burkitt's lymphoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human CD80 knockout Raji cell line (<a href='/en-us/products/cell-lines/human-cd80-knockout-raji-cell-line-ab273873'>ab273873</a>)

Lane 3:

Ramos (Human Burkitt's lymphoma cell line) whole cell lysate at 20 µg

Predicted band size: 33 kDa

Observed band size: 55 kDa

false