Anti-Scavenging Receptor SR-BI antibody [EPR20190]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Scavenging Receptor SR-BI antibody [EPR20190] (AB217318)

Immunohistochemical analysis of paraffin-embedded human liver tissue labeling Scavenging Receptor SR-BI with ab217318 at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous staining on human liver is observed. Counter stained with Hematoxylin.

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

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Scavenging Receptor SR-BI antibody [EPR20190] (AB217318)

Immunofluorescent analysis of 100% methanol fixed HepG2 (Human liver hepatocellular carcinoma cell line) cells labeling Scavenging Receptor SR-BI with ab217318 at 1/100 dilution followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining on HepG2 cell line.

The nuclear counter stain is DAPI (blue). Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594)) at 1/200 dilution (red).

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Scavenging Receptor SR-BI antibody [EPR20190] (AB217318)

Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma tissue labeling Scavenging Receptor SR-BI with ab217318 at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous staining on human hepatocellular carcinoma is observed. Counter stained with Hematoxylin.

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

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Scavenging Receptor SR-BI antibody [EPR20190] (AB217318)

Immunohistochemical analysis of paraffin-embedded human diffuse large B cell lymphoma tissue labeling Scavenging Receptor SR-BI with ab217318 at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous and cytoplasmic staining on human diffuse large B cell lymphoma is observed. Counter stained with Hematoxylin.

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

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IP

Supplier Data

Immunoprecipitation - Anti-Scavenging Receptor SR-BI antibody [EPR20190] (AB217318)

Scavenging Receptor SR-BI was immunoprecipitated from 0.35 mg of Human fetal liver lysate with ab217318 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab217318 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1 : Human fetal liver lysate, 10 μg (Input).

Lane 2 : ab217318 IP in Human fetal liver lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab217318 in Human fetal liver lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 30 seconds.

All lanes:

Immunoprecipitation - Anti-Scavenging Receptor SR-BI antibody [EPR20190] (ab217318)

Predicted band size: 60 kDa

Observed band size: 82 kDa

false

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Scavenging Receptor SR-BI antibody [EPR20190] (AB217318)

Immunohistochemical analysis of paraffin-embedded rat liver tissue labeling Scavenging Receptor SR-BI with ab217318 at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous and cytoplasmic staining on rat liver is observed. Counter stained with Hematoxylin.

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

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Scavenging Receptor SR-BI antibody [EPR20190] (AB217318)

Immunohistochemical analysis of paraffin-embedded rat cerebral cortex tissue labeling Scavenging Receptor SR-BI with ab217318 at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous and cytoplasmic staining on rat cerebral cortex blood vessel endothelium is observed. Counter stained with Hematoxylin.

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

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Scavenging Receptor SR-BI antibody [EPR20190] (AB217318)

Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling Scavenging Receptor SR-BI with ab217318 at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous staining on mouse liver is observed. Counter stained with Hematoxylin.

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

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• WB

Supplier Data

Western blot - Anti-Scavenging Receptor SR-BI antibody [EPR20190] (AB217318)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure times : Lane 1 : 30 seconds; Lane 2 : 4 seconds; Lane 3 : 2 seconds; Lane 4-5 : 5 seconds.

All lanes:

Western blot - Anti-Scavenging Receptor SR-BI antibody [EPR20190] (ab217318) at 1/2000 dilution

Lane 1:

U937 (Human histiocytic lymphoma cell line) whole cell lysate at 10 µg

Lane 2:

LNCaP (Human prostate cancer cell line) whole cell lysate at 10 µg

Lane 3:

PC-3 (Human prostate adenocarcinoma cell line) whole cell lysate at 10 µg

Lane 4:

THP-1 (Human monocytic leukemia cell line) whole cell lysate at 10 µg

Lane 5:

HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 10 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

Predicted band size: 60 kDa

Observed band size: 82 kDa

false

• WB

Lab

Western blot - Anti-Scavenging Receptor SR-BI antibody [EPR20190] (AB217318)

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 - ab217318 observed at 80 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab217318 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. ab217318 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.

All lanes:

Western blot - Anti-Scavenging Receptor SR-BI antibody [EPR20190] (ab217318)

Predicted band size: 60 kDa

false

• WB

Lab

Western blot - Anti-Scavenging Receptor SR-BI antibody [EPR20190] (AB217318)

False colour image of Western blot : Anti-Scavenging Receptor SR-BI antibody [EPR20190] staining at 1/2000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab217318 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 [EPR20190] (ab217318) at 1/2000 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

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• WB

Supplier Data

Western blot - Anti-Scavenging Receptor SR-BI antibody [EPR20190] (AB217318)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure times : Lane 1-3 : 4 seconds; Lane 4-6 : 3 minutes.

Scavenging Receptor SR-BI undergoes N-glycosylation when it is expressed. The molecular weight observed is consistent with what has been described in the literature (PMID : 12016218; PMID : 10821819; PMID : 224442701).

All lanes:

Western blot - Anti-Scavenging Receptor SR-BI antibody [EPR20190] (ab217318) at 1/2000 dilution

Lane 1:

Human fetal liver lysate at 20 µg

Lane 2:

Mouse liver lysate at 20 µg

Lane 3:

Rat liver lysate at 20 µg

Lane 4:

Mouse heart lysate at 10 µg

Lane 5:

Mouse kidney lysate at 10 µg

Lane 6:

Mouse spleen lysate at 10 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

Predicted band size: 60 kDa

Observed band size: 82 kDa

false

• WB

Supplier Data

Western blot - Anti-Scavenging Receptor SR-BI antibody [EPR20190] (AB217318)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Scavenging Receptor SR-BI antibody [EPR20190] (ab217318) at 1/2000 dilution

Lane 1:

C6 (Rat glial tumor cell line) whole cell lysate at 10 µg

Lane 2:

RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg

Lane 3:

PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate at 10 µg

Lane 4:

NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate at 10 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

Predicted band size: 60 kDa

Observed band size: 82 kDa

false

Exposure time: 30s