重组Anti-LIMPII抗体[EPR12080]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LIMPII antibody [EPR12080] (AB176317)

Immunohistochemical analysis of paraffin-embedded human kidney tissue labling LIMPII using ab176317 at a 1/50 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LIMPII antibody [EPR12080] (AB176317)

Immunohistochemical analysis of paraffin-embedded human liver tissue labling LIMPII using ab176317 at a 1/50 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IP

Supplier Data

Immunoprecipitation - Anti-LIMPII antibody [EPR12080] (AB176317)

Western blot analysis on immunoprecipitation pellet from Y79 (Human retinoblastoma cell line) cell lysate labeling LIMPII using ab176317 at a 1/10 dilution.

All lanes:

Immunoprecipitation - Anti-LIMPII antibody [EPR12080] (ab176317)

Predicted band size: 54 kDa

false

• WB

Lab

Western blot - Anti-LIMPII antibody [EPR12080] (AB176317)

False colour image of Western blot : Anti-LIMPII antibody [EPR12080] 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, ab176317 was shown to bind specifically to LIMPII. 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-LIMPII antibody [EPR12080] (ab176317) at 1/1000 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='/products/cell-lines/human-scarb2-knockout-mcf7-cell-line-ab274952'>ab274952</a>)

Lane 3:

SH-SY5Y cell lysate at 20 µg

Lane 4:

HEK-293 cell lysate at 20 µg

Predicted band size: 54 kDa

Observed band size: 80 kDa

false

• WB

Collaborator

Western blot - Anti-LIMPII antibody [EPR12080] (AB176317)

ab176317 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 ab176317 overnight at 4 °C at a 1/1000 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-LIMPII antibody [EPR12080] (ab176317) at 1/1000 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='/products/cell-lines/human-scarb2-knockout-mcf7-cell-line-ab274952'>ab274952</a>) at 30 µg

false

• WB

Supplier Data

Western blot - Anti-LIMPII antibody [EPR12080] (AB176317)

All lanes:

Western blot - Anti-LIMPII antibody [EPR12080] (ab176317) at 1/1000 dilution

Lane 1:

HeLa (Human epithelial cell line from cervix adenocarcinoma) lysate

Lane 2:

Y79 (Human retinoblastoma cell line) lysate

Lane 3:

Rat eyeball lysate

Lane 4:

Rat muscle lysate

Lane 5:

Mouse liver lysate

Predicted band size: 54 kDa

false