Anti-SFRP1 antibody [EPR7003]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SFRP1 antibody [EPR7003] (AB126613)

Unpurified ab126613, at 1/50 dilution, staining SFRP1 in Formalin-fixed, Paraffin-embedded Human heart muscle tissue by Immunohistochemistry.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SFRP1 antibody [EPR7003] (AB126613)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human testis tissue sections labeling SFRP1 with Purified ab126613 at 1 : 50 dilution (8 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1 : 0 dilution. PBS instead of the primary antibody was used as the negative control.

• WB

Lab

Western blot - Anti-SFRP1 antibody [EPR7003] (AB126613)

ab126613 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 ab126613 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 [EPR7003] (ab126613) 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

Observed band size: 35 kDa

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• WB

Lab

Western blot - Anti-SFRP1 antibody [EPR7003] (AB126613)

Blocking and diluting buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-SFRP1 antibody [EPR7003] (ab126613) at 1/5000 dilution

Lane 1:

Human fetal kidney lysates at 15 µg

Lane 2:

Human fetal heart lysates at 15 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 35 kDa

Observed band size: 36 kDa

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• WB

Lab

Western blot - Anti-SFRP1 antibody [EPR7003] (AB126613)

Blocking and diluting buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-SFRP1 antibody [EPR7003] (ab126613) at 1/1000 dilution

All lanes:

MDA-MB-435S (Human mammary gland ductal carcinoma melanocyte) whole cell lysates at 15 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 35 kDa

false

• WB

Unknown

Western blot - Anti-SFRP1 antibody [EPR7003] (AB126613)

All lanes:

Western blot - Anti-SFRP1 antibody [EPR7003] (ab126613) at 1/1000 dilution

Lane 1:

Human heart lysate at 10 µg

Lane 2:

MDA-MB-435 lysate at 10 µg

Lane 3:

A375 lysate at 10 µg

Secondary

All lanes:

Goat Anti-rabbit HRP at 1/2000 dilution

Predicted band size: 35 kDa

false

• OI-RD Scanning

Unknown

OI-RD Scanning - Anti-SFRP1 antibody [EPR7003] (AB126613)

We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody. Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.