Anti-TrkA antibody [EPR25641-50] (BSA and Azide free)

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-TrkA antibody [EPR25641-50] (BSA and Azide free) (AB302525)

This data was developed using ab302524, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized TF-1 (human Erythroleukemia erythroblast) cells labelling TrkA with ab302524 at 1/50 (10.82 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing membranous and cytoplasmic staining in TF-1 cell lineNegative control : 293T (PMID : 10523865) is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• IP

Supplier Data

Immunoprecipitation - Anti-TrkA antibody [EPR25641-50] (BSA and Azide free) (AB302525)

This data was developed using ab302524, the same antibody clone in a different buffer formulation.

TrkA was immunoprecipitated from 0.35 mg TF-1 (human Erythroleukemia erythroblast), whole cell lysate 10 µg with ab302524 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab302524 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

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

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

All lanes:

Immunoprecipitation - Anti-TrkA antibody [EPR25641-50] (<a href='/en-us/products/primary-antibodies/trka-antibody-epr25641-50-ab302524'>ab302524</a>) at 1/1000 dilution

Lane 1:

TF-1 (human Erythroleukemia erythroblast), whole cell lysate at 10 µg

Lane 2:

<a href='/en-us/products/primary-antibodies/trka-antibody-epr25641-50-ab302524'>ab302524</a> at 1/30 IP in TF-1 whole cell lysate at 10 µg

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of <a href='/en-us/products/primary-antibodies/trka-antibody-epr25641-50-ab302524'>ab302524</a> in TF-1 whole cell lysate at 10 µg

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Predicted band size: 87 kDa

Observed band size: 100 kDa,140 kDa

true

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-TrkA antibody [EPR25641-50] (BSA and Azide free) (AB302525)

This data was developed using ab302524, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 19330021, 8636219). Negative control : 293T (PMID : 10523865, 11251075) The blot of lane 2 + lane 3 was developed using a high sensitivity ECL substrate.

All lanes:

Western blot - Anti-TrkA antibody [EPR25641-50] (<a href='/en-us/products/primary-antibodies/trka-antibody-epr25641-50-ab302524'>ab302524</a>) at 1/1000 dilution

Lane 1:

TF-1 (human Erythroleukemia erythroblast), whole cell lysate 20 µg

Lane 2:

SH-SY5Y (human neuroblastoma epithelial cell), whole cell lysate 20 µg

Lane 3:

293T (human embryonic kidney epithelial cell), whole cell lysate 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 1/100000 dilution

Observed band size: 100 kDa,140 kDa

false

Exposure time: 70s

• WB

Supplier Data

Western blot - Anti-TrkA antibody [EPR25641-50] (BSA and Azide free) (AB302525)

This data was developed using ab302524, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST This antibody does not cross-react with human TrkB and TrkC.

All lanes:

Western blot - Anti-TrkA antibody [EPR25641-50] (<a href='/en-us/products/primary-antibodies/trka-antibody-epr25641-50-ab302524'>ab302524</a>) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney) transfected with an empty vector (vector control) containing a myc-His-tag®, whole cell lysate

Lane 2:

293T transfected with human TrkA expression vector containi a myc-His-tag®, whole cell lysate

Lane 3:

293T transfected with human TrkB expression vector containi a myc-His-tag®, whole cell lysate

Lane 4:

293T transfected with human TrkC expression vector containi a myc-His-tag®, whole cell lysate

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 1/100000 dilution

Observed band size: 75 kDa,150 kDa

false

Exposure time: 70s