Anti-Netrin 1 antibody [EPR5428] - BSA and Azide free

• IP

Supplier Data

Immunoprecipitation - Anti-Netrin 1 antibody [EPR5428] - BSA and Azide free (AB201324)

This data was developed using ab126729, the same antibody clone in a different buffer formulation.

Immunoprecipitation of NTN1 in MCF7 cell culture media. Culture medium was prepared and immunoprecipitation was performed using 2 μg of ab126729 pre-coupled to Protein A beads. Samples were then washed and processed for western blot.

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:

Immunoprecipitation - Anti-Netrin 1 antibody [EPR5428] (<a href='/en-us/products/primary-antibodies/netrin-1-antibody-epr5428-ab126729'>ab126729</a>) at 2 µg

false

• WB

Unknown

Western blot - Anti-Netrin 1 antibody [EPR5428] - BSA and Azide free (AB201324)

This data was developed using ab126729, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-Netrin 1 antibody [EPR5428] (<a href='/en-us/products/primary-antibodies/netrin-1-antibody-epr5428-ab126729'>ab126729</a>) at 1/1000 dilution

Lane 1:

Human fetal brain lysate at 10 µg

Lane 2:

Human small intestine lysate at 10 µg

Secondary

All lanes:

HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution

Predicted band size: 67 kDa

false

• WB

Lab

Western blot - Anti-Netrin 1 antibody [EPR5428] - BSA and Azide free (AB201324)

This data was developed using ab126729, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Netrin 1 antibody [EPR5428] (<a href='/en-us/products/primary-antibodies/netrin-1-antibody-epr5428-ab126729'>ab126729</a>) at 1/1000 dilution

All lanes:

Human fetal brain tissue 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 1/50000 dilution

Predicted band size: 67 kDa

Observed band size: 75 kDa

false

• WB

Supplier Data

Western blot - Anti-Netrin 1 antibody [EPR5428] - BSA and Azide free (AB201324)

This data was developed using ab126729, the same antibody clone in a different buffer formulation.

ab126729 was shown to react with NTN1 in wild-type MCF7 cell culture media in Western blot with loss of signal observed in culture media from NTN1 knockout cell line ab323856. Wild-type MCF7 and NTN1 knockout cell culture medium was subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab126729 overnight at 4 °C at a 1/2000 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-Netrin 1 antibody [EPR5428] (<a href='/en-us/products/primary-antibodies/netrin-1-antibody-epr5428-ab126729'>ab126729</a>) at 1/2000 dilution

Lane 1:

Wild-type MCF7 cell culture medium at 10 µg

Lane 2:

NTN1 knock-out MCF7 cell culture medium at 10 µg

false

• WB

Unknown

Western blot - Anti-Netrin 1 antibody [EPR5428] - BSA and Azide free (AB201324)

This data was developed using ab126729, the same antibody clone in a different buffer formulation.

Blocking/Diluting buffer and concentration : 5% NFDM/TBST

All lanes:

Western blot - Anti-Netrin 1 antibody [EPR5428] (<a href='/en-us/products/primary-antibodies/netrin-1-antibody-epr5428-ab126729'>ab126729</a>) at 1/500 dilution

Lane 1:

Rat brain lysates prepared in RIPA lysis method at 20 µg

Lane 2:

Rat brain lysates prepared in 1%SDS Hot lysis method 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: 67 kDa

Observed band size: 75 kDa

true

Exposure time: 180s

• WB

Lab

Western blot - Anti-Netrin 1 antibody [EPR5428] - BSA and Azide free (AB201324)

This data was developed using ab126729, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Netrin 1 antibody [EPR5428] (<a href='/en-us/products/primary-antibodies/netrin-1-antibody-epr5428-ab126729'>ab126729</a>) at 1/1000 dilution

Lane 1:

Mouse kidney tissue lysate at 20 µg

Lane 2:

Mouse heart tissue lysate at 20 µg

Lane 3:

Rat kidney tissue 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 1/50000 dilution

Predicted band size: 67 kDa

Observed band size: 75 kDa

false