Anti-Netrin 1 antibody [EPR5428]

• IP

Supplier Data

Immunoprecipitation - Anti-Netrin 1 antibody [EPR5428] (AB126729)

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] (ab126729) at 2 µg

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• WB

Supplier Data

Western blot - Anti-Netrin 1 antibody [EPR5428] (AB126729)

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] (ab126729) 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

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• WB

Lab

Western blot - Anti-Netrin 1 antibody [EPR5428] (AB126729)

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Netrin 1 antibody [EPR5428] (ab126729) 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

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• WB

Unknown

Western blot - Anti-Netrin 1 antibody [EPR5428] (AB126729)

All lanes:

Western blot - Anti-Netrin 1 antibody [EPR5428] (ab126729) 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

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• WB

Lab

Western blot - Anti-Netrin 1 antibody [EPR5428] (AB126729)

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Netrin 1 antibody [EPR5428] (ab126729) 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

• WB

Unknown

Western blot - Anti-Netrin 1 antibody [EPR5428] (AB126729)

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

All lanes:

Western blot - Anti-Netrin 1 antibody [EPR5428] (ab126729) 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

CiteAb

Western blot - Anti-Netrin 1 antibody [EPR5428] (AB126729)

Netrin 1 western blot using anti-Netrin 1 antibody [EPR5428] ab126729. Publication image and figure legend from Hadi, T., Boytard, L., et al., 2018, Nat Commun, PubMed 30479344.

ab126729 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab126729 please see the product overview.

Netrin-1 is highly expressed in human and murine AAA. a Quantitative PCR analysis of Ntn1, Adgre1, Ccl2, and Il6 mRNA levels isolated from aortas of ApoE-/- mice exposed to PBS or Ang II for 28 days (n = 8/9 mice per group). *p < 0.05, ***p < 0.001. b Western blot analysis of netrin-1 in aorta extracts normalized to loading control, GAPDH (n = 3 mice per group). AU arbitrary units. ** p < 0.01. Representative microscopy staining of netrin-1 and cell nuclei (DAPI, blue) in magnified aortic sections (dashed box) from PBS and Ang II infused ApoE-/- mice (c) (whole aortic section is shown in left top and bottom, scale bar 200 μm, magnified areas on the right, scale bar 20 μm) and human specimens (d); Hematoxylin and eosin (H&E; scale bar 500 μm) staining showing magnified sections (1, 2; scale bar 20 μm); arrows indicate acellular localization of netrin-1; L lumen, A adventitia. Unpaired, two-tailed t-test was used for statistical analysis. Error bars represent s.e.m

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