Anti-Scavenging Receptor SRB2 antibody [EPR12081] - BSA and Azide free

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Scavenging Receptor SRB2 antibody [EPR12081] - BSA and Azide free (AB251221)

This data was developed using ab196651, the same antibody clone in a different buffer formulation.

Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed MCF7 (Human breast adenocarcinoma cell line) cells labeling Scavenging Receptor SRB2 with ab196651 at 1/150 dilution (red) compared with a rabbit monoclonal IgG isotype control (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Scavenging Receptor SRB2 antibody [EPR12081] - BSA and Azide free (AB251221)

This data was developed using ab196651, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling Scavenging Receptor SRB2 with ab196651 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Cytoplasm staining on human liver tissue is observed. Counter stained with Hematoxylin. Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Negative control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Scavenging Receptor SRB2 antibody [EPR12081] - BSA and Azide free (AB251221)

This data was developed using ab196651, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized MCF7 (human breast adenocarcinoma epithelial cell) cells labelling Scavenging Receptor SRB2 with ab196651 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed (ab150081) secondary antibody at 1/1000 dilution.

Confocal image showing cytoplasmic staining in MCF7 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Magenta).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Scavenging Receptor SRB2 antibody [EPR12081] - BSA and Azide free (AB251221)

This data was developed using ab196651, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized Neuro-2a (mouse neuroblastoma neuroblast) cells labelling Scavenging Receptor SRB2 with ab196651 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed (ab150081) secondary antibody at 1/1000 dilution.

Confocal image showing cytoplasmic staining in Neuro-2a cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Magenta).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Scavenging Receptor SRB2 antibody [EPR12081] - BSA and Azide free (AB251221)

This data was developed using ab196651, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling Scavenging Receptor SRB2 with ab196651 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Cytoplasm staining on mouse liver tissue is observed. Counter stained with Hematoxylin. Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Negative control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

• WB

Lab

Western blot - Anti-Scavenging Receptor SRB2 antibody [EPR12081] - BSA and Azide free (AB251221)

This data was developed using ab196651, the same antibody clone in a different buffer formulation.

ab196651 was shown to react with SCARB2 in wild-type MCF7 cells in Western blot with loss of signal observed in SCARB2 knockout cell line ab274952. Wild-type MCF7 and SCARB2 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab196651 overnight at 4 °C at a 1/20000 dilution. Blots were incubated with secondary antibodies at 0.2 µg/mL before imaging.

This data was provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

All lanes:

Western blot - Anti-Scavenging Receptor SRB2 antibody [EPR12081] - C-terminal (<a href='/en-us/products/primary-antibodies/scavenging-receptor-srb2-antibody-epr12081-c-terminal-ab196651'>ab196651</a>) at 1/20000 dilution

Lane 1:

Wild-type MCF7 lysate at 30 µg

Lane 2:

SCARB2 knock-out MCF7 lysate at 30 µg

Lane 2:

Western blot - Human SCARB2 knockout MCF7 cell line (<a href='/en-us/products/cell-lines/human-scarb2-knockout-mcf7-cell-line-ab274952'>ab274952</a>) at 30 µg

false

• WB

Lab

Western blot - Anti-Scavenging Receptor SRB2 antibody [EPR12081] - BSA and Azide free (AB251221)

False colour image of Western blot : Anti-Scavenging Receptor SRB2 antibody [EPR12081] - C-terminal staining at 1/20000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab196651 was shown to bind specifically to Scavenging Receptor SRB2. A band was observed at 80 kDa in wild-type MCF7 cell lysates with no signal observed at this size in SCARB2 knockout cell line ab274952 (knockout cell lysate ab275010). To generate this image, wild-type and SCARB2 knockout MCF7 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 SRB2 antibody [EPR12081] - C-terminal (<a href='/en-us/products/primary-antibodies/scavenging-receptor-srb2-antibody-epr12081-c-terminal-ab196651'>ab196651</a>) at 1/20000 dilution

Lane 1:

Wild-type MCF7 cell lysate at 20 µg

Lane 2:

SCARB2 knockout MCF7 cell lysate at 20 µg

Lane 2:

Western blot - Human SCARB2 knockout MCF7 cell line (<a href='/en-us/products/cell-lines/human-scarb2-knockout-mcf7-cell-line-ab274952'>ab274952</a>)

Predicted band size: 54 kDa

Observed band size: 80 kDa

false

• WB

Supplier Data

Western blot - Anti-Scavenging Receptor SRB2 antibody [EPR12081] - BSA and Azide free (AB251221)

This data was developed using ab196651, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

The observed MW is consistent with what has been described in the literature (PMID : 21389126; 22272359; 23635510).

All lanes:

Western blot - Anti-Scavenging Receptor SRB2 antibody [EPR12081] - C-terminal (<a href='/en-us/products/primary-antibodies/scavenging-receptor-srb2-antibody-epr12081-c-terminal-ab196651'>ab196651</a>) at 1/20000 dilution

All lanes:

Human fetal kidney tissue lysate at 10 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 54 kDa

Observed band size: 80 kDa

false

• WB

Supplier Data

Western blot - Anti-Scavenging Receptor SRB2 antibody [EPR12081] - BSA and Azide free (AB251221)

This data was developed using ab196651, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

The observed MW is consistent with what has been described in the literature (PMID : 21389126, 22272359, and 23635510).

All lanes:

Western blot - Anti-Scavenging Receptor SRB2 antibody [EPR12081] - C-terminal (<a href='/en-us/products/primary-antibodies/scavenging-receptor-srb2-antibody-epr12081-c-terminal-ab196651'>ab196651</a>) at 1/20000 dilution

Lane 1:

MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 10 µg

Lane 2:

HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 54 kDa

Observed band size: 80 kDa

false