Anti-RIP2 antibody [EPR26079-37] - BSA and Azide free

• WB

Supplier Data

Western blot - Anti-RIP2 antibody [EPR26079-37] - BSA and Azide free (AB303555)

Blocking and dilution buffer and concentration : ½ volume of Odyssey Blocking Buffer (TBS)+ ½ volume of 0.1% TBS.  False colour image of Western blot : Anti-RIP2 antibody [EPR26079-37](ab303554) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab303554 was shown to bind specifically to RIP2. A band was observed at 65 kDa in wild-type HeLa cell lysates with no signal observed at this size in RIP2 knockout cell line ab264688 (knockout cell lysate ab258636). To generate this image, wild-type and RIP2 knockout HeLa cell lysates were analyzed. First, samples were run on an SDS-PAGE gel then transferred onto an immobilon-FL PVDF membrane. Membranes were blocked in in Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-RIP2 antibody [EPR26079-37] (<a href='/en-us/products/primary-antibodies/rip2-antibody-epr26079-37-ab303554'>ab303554</a>)

Lane 1:

Wild-type HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

RIP2 knockout HeLa whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-800cw-preadsorbed-ab216772'>ab216772</a>) at 1/20000 dilution

false

• WB

Supplier Data

Western blot - Anti-RIP2 antibody [EPR26079-37] - BSA and Azide free (AB303555)

This data was developed using ab303554, the same antibody clone in a different buffer formulation. Blocking and dilution buffer and concentration : 5% NFDM/TBST.

All lanes:

Western blot - Anti-RIP2 antibody [EPR26079-37] (<a href='/en-us/products/primary-antibodies/rip2-antibody-epr26079-37-ab303554'>ab303554</a>) at 1/1000 dilution

All lanes:

Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 61 kDa

Observed band size: 61 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-RIP2 antibody [EPR26079-37] - BSA and Azide free (AB303555)

This data was developed using ab303554, the same antibody clone in a different buffer formulation. Blocking and dilution buffer and concentration : 5% NFDM/TBST. Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

All lanes:

Western blot - Anti-RIP2 antibody [EPR26079-37] (<a href='/en-us/products/primary-antibodies/rip2-antibody-epr26079-37-ab303554'>ab303554</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

HEK-293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 3:

THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg

Lane 4:

Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 61 kDa

Observed band size: 61 kDa

false

Exposure time: 180s