Anti-DR5 antibody [EPR22276] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DR5 antibody [EPR22276] - BSA and Azide free (AB243847)

Immunohistochemical analysis of paraffin-embedded human colon cancer tissue labeling DR5 with ab230969 at 1/1000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Membranous and cytoplasmic staining in human colon cancer cells (PMID : 23284732) is observed. Counterstained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab230969).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DR5 antibody [EPR22276] - BSA and Azide free (AB243847)

Immunohistochemical analysis of paraffin-embedded human endometrium tissue labeling DR5 with ab230969 at 1/2000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining in endothelial cells of human endometrium (PMID : 22789540, 12874246, 15987601) is observed. Counterstained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab230969).

• IP

Supplier Data

Immunoprecipitation - Anti-DR5 antibody [EPR22276] - BSA and Azide free (AB243847)

DR5 was immunoprecipitated from 0.35 mg U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate with ab230969 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab230969 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.
Lane 1 : U-87 MG whole cell lysate 10 μg (Input).
Lane 2 : ab230969 IP in U-87 MG whole cell lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab230969 in U-87 MG whole cell lysate.
Blocking/Dilution buffer : 5% NFDM/TBST.
Exposure time : 20 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab230969).

All lanes:

Immunoprecipitation - Anti-DR5 antibody [EPR22276] (<a href='/ko/products/primary-antibodies/dr5-antibody-epr22276-ab230969'>ab230969</a>)

Predicted band size: 47 kDa

Observed band size: 40 kDa,48 kDa

false

• WB

Lab

Western blot - Anti-DR5 antibody [EPR22276] - BSA and Azide free (AB243847)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab230969).

Western blot : Anti-DR5 antibody [EPR22276] ab230969 staining at 1/1000 dilution, shown in green; Mouse anti-CANX ab238078 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 37-50 kDa in Wild-type HeLa cell lysates with no signal observed at this size in TNFRSF10B knockout HeLa cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5% 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-DR5 antibody [EPR22276] (<a href='/ko/products/primary-antibodies/dr5-antibody-epr22276-ab230969'>ab230969</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysates at 20 µg

Lane 2:

Western blot - Human TNFRSF10B (DR5) knockout HeLa cell line (<a href='/ko/products/cell-lines/human-tnfrsf10b-dr5-knockout-hela-cell-line-ab264922'>ab264922</a>) at 20 µg

Lane 3:

HepG2 cell lysates at 20 µg

Lane 4:

HCT 116 cell lysates at 20 µg

Secondary

All lanes:

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

Predicted band size: 47 kDa

Observed band size: 37-50 kDa

false