Anti-htrA1 antibody [EPR23240-64] - BSA and Azide free

• WB

Lab

Western blot - Anti-htrA1 antibody [EPR23240-64] - BSA and Azide free (AB274327)

ab274322 was shown to react with HTRA1 in wild-type HAP1 cells in Western blot with loss of signal observed in a HTRA1 knockout cell line. Wild-type HAP1 and HTRA1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab274322 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-htrA1 antibody [EPR23240-64] (<a href='/en-us/products/primary-antibodies/htra1-antibody-epr23240-64-ab274322'>ab274322</a>) at 1/1000 dilution

Lane 1:

Wild-type HAP1 lysate at 20 µg

Lane 2:

HTRA1 Knockout HAP1 lysate at 20 µg

Observed band size: 51 kDa

false

• IP

Unknown

Immunoprecipitation - Anti-htrA1 antibody [EPR23240-64] - BSA and Azide free (AB274327)

htrA1 was immunoprecipitated from 0.35 mg Human placenta lysate with ab274322 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab274322 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Human placenta lysate 10 ug

Lane 2 : ab274322 IP in Human placenta lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab274322 in Human placenta lysate

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

Exposure time : 7.75 seconds

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab274322).

All lanes:

Immunoprecipitation - Anti-htrA1 antibody [EPR23240-64] (<a href='/en-us/products/primary-antibodies/htra1-antibody-epr23240-64-ab274322'>ab274322</a>)

Predicted band size: 51 kDa

Observed band size: 51 kDa

false

• WB

Lab

Western blot - Anti-htrA1 antibody [EPR23240-64] - BSA and Azide free (AB274327)

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

The expression profile observed is consistent with what has been described in the literature (PMID : 16767218, 19103920).

Exposure time : 48 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab274322).

All lanes:

Western blot - Anti-htrA1 antibody [EPR23240-64] (<a href='/en-us/products/primary-antibodies/htra1-antibody-epr23240-64-ab274322'>ab274322</a>) at 1/1000 dilution

Lane 1:

Human brain tissue lysate at 20 µg

Lane 2:

Human placenta tissue lysate at 20 µg

Lane 3:

Human ovary tissue lysate at 20 µg

Lane 4:

Human ovary cancer tissue lysate at 20 µg

Secondary

All lanes:

VeriBlot for IP secondary antibody(HRP)(<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Predicted band size: 51 kDa

Observed band size: 51 kDa

false

• WB

Lab

Western blot - Anti-htrA1 antibody [EPR23240-64] - BSA and Azide free (AB274327)

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

Low expression : HepG2, MCF7 (PMID : 14716297, 22761798).

Exposure time : 3 minutes.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab274322).

All lanes:

Western blot - Anti-htrA1 antibody [EPR23240-64] (<a href='/en-us/products/primary-antibodies/htra1-antibody-epr23240-64-ab274322'>ab274322</a>) at 1/1000 dilution

Lane 1:

SK-OV-3 (human ovarian cancer epithelial cell), whole cell lysate at 20 µg

Lane 2:

HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate at 20 µg

Lane 3:

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

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 51 kDa

Observed band size: 51 kDa

false