Anti-Scavenging Receptor SR-BI antibody [EP1556Y] - Low endotoxin, Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Scavenging Receptor SR-BI antibody [EP1556Y] - Low endotoxin, Azide free (AB176238)

Immunohistochemical staining of paraffin embedded human liver with purified ab52629 at a working dilution of 1/500. The secondary antibody used is ab97051, a goat anti-rabbit IgG (H&L) at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52629).

• WB

Lab

Western blot - Anti-Scavenging Receptor SR-BI antibody [EP1556Y] - Low endotoxin, Azide free (AB176238)

False colour image of Western blot : Anti-Scavenging Receptor SR-BI antibody [EP1556Y] 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, ab52629 was shown to bind specifically to Scavenging Receptor SR-BI. A band was observed at 70/75 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in SCARB1 knockout cell line ab282646 (knockout cell lysate ab283046). To generate this image, wild-type and SCARB1 knockout HEK-293T cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % 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 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-Scavenging Receptor SR-BI antibody [EP1556Y] (<a href='/en-us/products/primary-antibodies/scavenging-receptor-sr-bi-antibody-ep1556y-ab52629'>ab52629</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

SCARB1 knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human SCARB1 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-scarb1-knockout-hek-293t-cell-line-ab282646'>ab282646</a>)

Lane 3:

Human Liver cell lysate at 20 µg

Predicted band size: 60 kDa

Observed band size: 70 kDa,75 kDa

false

• WB

Lab

Western blot - Anti-Scavenging Receptor SR-BI antibody [EP1556Y] - Low endotoxin, Azide free (AB176238)

Lane 1 : Wild-type HAP1 whole cell lysate (20 μg)
Lane 2 : Scavenging Receptor SR-BI knockout HAP1 whole cell lysate (20 μg)
Lane 3 : HepG2 whole cell lysate (20 μg)
Lane 4 : Human liver whole cell lysate (20 μg)

Lanes 1 - 4 : Merged signal (red and green). Green - ab52629 observed at 80 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab52629 was shown to specifically react with Scavenging Receptor SR-BI in wild-type HAP1 cells as signal was lost in Scavenging Receptor SR-BI knockout cells. Wild-type and Scavenging Receptor SR-BI knockout samples were subjected to SDS-PAGE. ab52629 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/2000 dilution and 1/20000 dilution respectively. Blots were developed 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 1/20000 dilution for 1 hour at room temperature before imaging.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab176238).

All lanes:

Western blot - Anti-Scavenging Receptor SR-BI antibody [EP1556Y] (<a href='/en-us/products/primary-antibodies/scavenging-receptor-sr-bi-antibody-ep1556y-ab52629'>ab52629</a>)

Predicted band size: 60 kDa

false