Anti-B7-H6 antibody [EPR21841]

• WB

Lab

Western blot - Anti-B7-H6 antibody [EPR21841] (AB219205)

Western blot : Anti-B7-H6 antibody [EPR21841] ab219205 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 70-80 kDa in Wild-type Raji UNBOILED cell lysates with no signal observed at this size in NCR3LG1 knockout Raji UNBOILED cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5pc Milk in TBS-0.1 % Tween^™ 20 (TBS-T) 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 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-B7-H6 antibody [EPR21841] (ab219205) at 1/1000 dilution

Lane 1:

Wild-type Raji UNBOILED whole cell lysate at 20 µg

Lane 2:

Western blot - Human NCR3LG1 knockout Raji cell line (<a href='/ko/products/cell-lines/human-ncr3lg1-knockout-raji-cell-line-ab290414'>ab290414</a>) at 20 µg

Lane 3:

HepG2 UNBOILED whole cell lysate at 20 µg

Lane 4:

K562 UNBOILED whole cell lysate at 20 µg

Lane 5:

Daudi UNBOILED whole cell lysate at 20 µg

Lane 6:

SK-OV-3 UNBOILED whole cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 51 kDa

Observed band size: 70-80 kDa

false

• WB

Supplier Data

Western blot - Anti-B7-H6 antibody [EPR21841] (AB219205)

Blocking/Dilution buffer : 5% NFDM/TBST.

MCF7 cells have a low level of B7-H6 expression. The molecular mass observed is consistent with the literature (PMID 24780758).

All lanes:

Western blot - Anti-B7-H6 antibody [EPR21841] (ab219205) at 1/1000 dilution

Lane 1:

K562 (human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate at 20 µg

Lane 2:

MCF7 (human breast adenocarcinoma cell line) whole cell lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 51 kDa

Observed band size: 80 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-B7-H6 antibody [EPR21841] (AB219205)

Exposure time : Lanes 1 and 2 : 92 seconds; Lanes 3 and 4 : 3 minutes.

Blocking/Dilution buffer : 5% NFDM/TBST.

B7-H6 is a glycoprotein and can be de-glycosylated by PNGase F. The molecular mass observed is consistent with the literature (PMID 24780758).

All lanes:

Western blot - Anti-B7-H6 antibody [EPR21841] (ab219205) at 1/1000 dilution

Lane 1:

Untreated HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

HepG2 whole cell lysate treated with PNGase F at 20 µg

Lane 3:

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

Lane 4:

HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 51 kDa

Observed band size: 50-80 kDa

false