Anti-SFRP1 antibody [EPR23092-253]

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-SFRP1 antibody [EPR23092-253] (AB267466)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized MCF7 (Human breast adenocarcinoma epithelial cell, Left) treated with 5μg/ml BFA for 2h / SK-MEL-28 (Human malignant melanoma, Right) treated with 5μg/ml BFA for 2h labeling SFRP1 with ab267466 at 1/500 (Red) compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 was used as the secondary antibody.

Negative control : MCF7 (PMID : 14968126).

• IP

Supplier Data

Immunoprecipitation - Anti-SFRP1 antibody [EPR23092-253] (AB267466)

SFRP1 was immunoprecipitated from 0.35 mg human kidney lysate with ab267466 at 1/30 dilution (2μg in 0.35 mg lysates). Western blot was performed on the immunoprecipitate using ab267466 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : Human kidney lysate 10μg

Lane 2 : ab267466 IP in human kidney lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab267466 in human kidney lysate

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

Exposure time : 30 seconds.

All lanes:

Immunoprecipitation - Anti-SFRP1 antibody [EPR23092-253] (ab267466)

Predicted band size: 35 kDa

false

• IP

Unknown

Immunoprecipitation - Anti-SFRP1 antibody [EPR23092-253] (AB267466)

SFRP1 was immunoprecipitated from 0.35 mg mouse kidney lysate with ab267466 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab267466 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : Mouse kidney lysate 10μg

Lane 2 : ab267466 IP in mouse kidney lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab267466 in mouse kidney lysate

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

Exposure time : 3 mins.

All lanes:

Immunoprecipitation - Anti-SFRP1 antibody [EPR23092-253] (ab267466)

Predicted band size: 35 kDa

false

• WB

Lab

Western blot - Anti-SFRP1 antibody [EPR23092-253] (AB267466)

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

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 22927647, 11741940).

Negative control : MCF7 (PMID : 15289865).

Exposure time : 92 seconds.

All lanes:

Western blot - Anti-SFRP1 antibody [EPR23092-253] (ab267466) at 1/1000 dilution

Lane 1:

Human breast lysate at 20 µg

Lane 2:

Human kidney lysate at 20 µg

Lane 3:

Human heart lysate at 20 µg

Lane 4:

SK-MEL-28 (human malignant melanoma) whole cell lysate at 20 µg

Lane 5:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 6:

Mouse kidney lysate at 20 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 1/20000 dilution

Predicted band size: 35 kDa

Observed band size: 35 kDa

false

• WB

Lab

Western blot - Anti-SFRP1 antibody [EPR23092-253] (AB267466)

ab267466 was shown to react with SFRP1 in wild-type A549 cells in Western blot with loss of signal observed in SFRP1 knockout cell line ab277906. Wild-type A549 and SFRP1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab267466 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with secondary antibodies at 0.2ug/mL before imaging.

These data were 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-SFRP1 antibody [EPR23092-253] (ab267466) at 1/1000 dilution

Lanes 1 - 2:

Western blot - Human SFRP1 knockout A549 cell line (<a href='/en-us/products/cell-lines/human-sfrp1-knockout-a549-cell-line-ab277906'>ab277906</a>)

Lane 1:

Wild-type A549 lysate at 20 µg

Lane 2:

SFRP1 Knockout A549 lysate at 20 µg

false

• WB

Lab

Western blot - Anti-SFRP1 antibody [EPR23092-253] (AB267466)

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

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 22927647, 11741940).

Exposure time : 92 seconds.

All lanes:

Western blot - Anti-SFRP1 antibody [EPR23092-253] (ab267466) at 1/1000 dilution

Lane 1:

Rat kidney lysate at 20 µg

Lane 2:

NIH/3T3 (mouse embryonic fibroblast) 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 1/20000 dilution

Predicted band size: 35 kDa

Observed band size: 35 kDa

false

• WB

Lab

Western blot - Anti-SFRP1 antibody [EPR23092-253] (AB267466)

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

Exposure time : 114 seconds.

All lanes:

Western blot - Anti-SFRP1 antibody [EPR23092-253] (ab267466) at 1/5000 dilution

All lanes:

Recombinant human sFRP-1 protein, 20 ng

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: 35 kDa

Observed band size: 35 kDa

false