Anti-Scavenging Receptor SR-BI 抗体 [25/CLA-1] (ab300632)

False colour image of Western blot: Anti-Scavenging Receptor SR-BI antibody [25/CLA-1] staining at 1/1000 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab300632 was shown to bind specifically to Scavenging Receptor SR-BI. A band was observed at 70 and 75 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in SCARB1 knockout cell line ab282646. 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-Mouse IgG H&L (IRDye^® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed (ab216777) at 1/20000 dilution.

Blocking / Diluting buffer and concentration: 5% NFDM/TBST

False colour image of Western blot: Anti-Scavenging Receptor SR-BI antibody [725/CLA-1] (ab300632) staining at 1/1000 dilution, shown in green; Rabbit anti-GAPDH antibody [16891] (ab181602) loading control staining at 1/20000 dilution, shown in red.

In Western blot, ab300632 was shown to bind specifically to SCARB1. A band was observed at 80 kDa in wild-type HAP1 cell lysates with no signal observed at this size in the SCARB1 knockout cell line. To generate this image, wild-type and SCARB1 knockout HAP1 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a PVDF-FL membrane. Membranes were blocked in Odyssey diluted in 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. 

Performed under reducing conditions.

Immunohistochemical analysis of paraffin-embedded human liver labeling SCARB1 with ab300632 at 1/1000 dilution (1.01 µg/ml) followed by anti-mouse IgG1 antibody (ab125913) for 8 minutes during the Leica DS9800 kit staining procedure. Positive staining on human liver. The section was incubated with ab300632 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody only without primary.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

Immunohistochemical analysis of paraffin-embedded human cerebrum labeling SCARB1 with ab300632 at 1/1000 dilution (1.01 µg/ml) followed by anti-mouse IgG1 antibody (ab125913) for 8 minutes during the Leica DS9800 kit staining procedure. Positive staining on blood vessels of human cerebrum. The section was incubated with ab300632 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody only without primary.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma labeling SCARB1 with ab300632 at 1/1000 dilution (1.01 µg/ml) followed by anti-mouse IgG1 antibody (ab125913) for 8 minutes during the Leica DS9800 kit staining procedure. Positive staining on human hepatocellular carcinoma. The section was incubated with ab300632 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody only without primary.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins