Anti-ROR2 antibody [EPR27134-21] - BSA and Azide free

• WB

Supplier Data

Western blot - Anti-ROR2 antibody [EPR27134-21] - BSA and Azide free (AB309484)

This data was developed using ab309483, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST Low expression : 293T(PMID : 25542006) Lysates were freshly made and used for Western blotting immediately to minimize protein degradation. In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : Lane 1 : 180 seconds, Lane 2-3 : 37 seconds, Lane 3 : 114 seconds

All lanes:

Western blot - Anti-ROR2 antibody [EPR27134-21] (<a href='/en-us/products/primary-antibodies/ror2-antibody-epr27134-21-ab309483'>ab309483</a>) at 1/1000 dilution

Lane 1:

Saos-2 (human osteosarcoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

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

Lane 3:

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

Lane 4:

NIH/3T3 (mouse embryonic fibroblast) 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

Observed band size: 125 kDa

false

• WB

Supplier Data

Western blot - Anti-ROR2 antibody [EPR27134-21] - BSA and Azide free (AB309484)

This data was developed using ab309483, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : Intercept® (TBS) Blocking Buffer diluted with an equal volume of TBS Lysates at 20 µg per lane. The samples were run on a Bis-Tris gel under reducing conditions. Western blot : Anti-ROR2 antibody (ab309483) 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, ab309483 was shown to bind specifically to ROR2. Target of interest was observed at 125 kDa in wild-type HAP1 cell lysates (lane 1) with no signal observed at this size in ROR2 knockout cell line (lane 2). To generate this image, samples were first run on an SDS-PAGE gel then transferred onto an immobilon-FL PVDF membrane. Membranes were blocked in a fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4 °C. Blots were washed in TBS-T, incubated with secondary antibodies Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution for 1 h at room temperature, washed again then imaged.

Lane 1:

Wild-type HAP1(human chronic myelogenous leukemia near-haploid cell line) whole cell lysate at 20 µg

Lane 2:

ROR2 knockout HAP1 whole cell lysate

Observed band size: 125 kDa

false