Anti-SFRP1 antibody [EPR23092-253] - BSA and Azide free

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-SFRP1 antibody [EPR23092-253] - BSA and Azide free (AB269949)

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).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab267466).

• IP

Supplier Data

Immunoprecipitation - Anti-SFRP1 antibody [EPR23092-253] - BSA and Azide free (AB269949)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab267466).

All lanes:

Immunoprecipitation - Anti-SFRP1 antibody [EPR23092-253] (<a href='/en-us/products/primary-antibodies/sfrp1-antibody-epr23092-253-ab267466'>ab267466</a>)

Predicted band size: 35 kDa

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• IP

Unknown

Immunoprecipitation - Anti-SFRP1 antibody [EPR23092-253] - BSA and Azide free (AB269949)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab267466).

All lanes:

Immunoprecipitation - Anti-SFRP1 antibody [EPR23092-253] (<a href='/en-us/products/primary-antibodies/sfrp1-antibody-epr23092-253-ab267466'>ab267466</a>)

Predicted band size: 35 kDa

false

• WB

Lab

Western blot - Anti-SFRP1 antibody [EPR23092-253] - BSA and Azide free (AB269949)

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] (<a href='/en-us/products/primary-antibodies/sfrp1-antibody-epr23092-253-ab267466'>ab267466</a>) 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