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.

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.

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.

All lanes : 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 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 60 kDa
Observed band size: 82 kDa why is the actual band size different from the predicted?

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).

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

Lane 1 : U937 (Human histiocytic lymphoma cell line) whole cell lysate
Lane 2 : LNCaP (Human prostate cancer cell line) whole cell lysate
Lane 3 : PC-3 (Human prostate adenocarcinoma cell line) whole cell lysate
Lane 4 : THP-1 (Human monocytic leukemia cell line) whole cell lysate
Lane 5 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 60 kDa
Observed band size: 82 kDa why is the actual band size different from the predicted?

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

Lane 1 : C6 (Rat glial tumor cell line) whole cell lysate
Lane 2 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
Lane 4 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 60 kDa
Observed band size: 82 kDa why is the actual band size different from the predicted?


Exposure time: 30 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.

 

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.

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.

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.

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.

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.