Anti-TrkB antibody [EPR17805-146] - Low endotoxin, Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TrkB antibody [EPR17805-146] - Low endotoxin, Azide free (AB223155)

Immunohistochemical analysis of paraffin-embedded mouse cerebral cortex tissue labeling TrkB with ab187041 at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Cytoplasmic and membrane staining on mouse cerebral cortex is observed.

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS BSA glycerol and sodium azide (ab187041).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TrkB antibody [EPR17805-146] - Low endotoxin, Azide free (AB223155)

Immunohistochemical analysis of paraffin-embedded rat cerebral cortex tissue labeling TrkB with ab187041 at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Cytoplasmic and membrane staining on rat cerebral cortex is observed.

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS BSA glycerol and sodium azide (ab187041).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-TrkB antibody [EPR17805-146] - Low endotoxin, Azide free (AB223155)

This data was developed using ab187041, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neuron cells labelling TrkB with ab187041 at 1/50 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing positive staining in mouse primary neuron.Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection. ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 4 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 2 ug/ml dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TrkB antibody [EPR17805-146] - Low endotoxin, Azide free (AB223155)

Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling TrkB with ab187041 at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Negative control : no staining on mouse liver [PMID : 2555172].

Counter stained with Hematoxylin.

This data was developed using the same antibody clone in a different buffer formulation containing PBS BSA glycerol and sodium azide (ab187041).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IP

Supplier Data

Immunoprecipitation - Anti-TrkB antibody [EPR17805-146] - Low endotoxin, Azide free (AB223155)

TrkB was immunoprecipitated from 1 mg of mouse brain lysate with ab187041 at 1/50 dilution.

Western blot was performed from the immunoprecipitate using ab187041 at 1/1000 dilution.

VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1 : Mouse brain lysate 10μg (Input).

Lane 2 : ab187041 IP in mouse brain lysate.

Lane 3 : Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab187041 in mouse brain lysate.

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

Exposure time : 1 second.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab187041).

All lanes:

Immunoprecipitation - Anti-TrkB antibody [EPR17805-146] (<a href='/en-us/products/primary-antibodies/trkb-antibody-epr17805-146-ab187041'>ab187041</a>)

Predicted band size: 92 kDa

false

• I-ELISA

Lab

Indirect ELISA - Anti-TrkB antibody [EPR17805-146] - Low endotoxin, Azide free (AB223155)

This data was developed using ab223155, the same antibody clone in a different buffer formulation.

ELISA analysis of Mouse TrkB (Ntrk2) recombinant protein at 1000 ng/ml with ab187041. An Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/2500 dilution was used as the secondary antibody.

• WB

Supplier Data

Western blot - Anti-TrkB antibody [EPR17805-146] - Low endotoxin, Azide free (AB223155)

This data was developed using ab187041, the same clone in a different buffer formulation.

Blocking/Dilution buffer : 5% NFDM/TBST.

Mouse TrkB active protein contains aa1-429 with a C-terminal poly histidine-tag.

All lanes:

Western blot - Anti-TrkB antibody [EPR17805-146] (<a href='/en-us/products/primary-antibodies/trkb-antibody-epr17805-146-ab187041'>ab187041</a>) at 1/5000 dilution

All lanes:

Mouse TrkB active protein at 0.01 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

Predicted band size: 171 kDa,87 kDa,92 kDa

Observed band size: 171 kDa,75-85 kDa

false

Exposure time: 3s

• WB

Supplier Data

Western blot - Anti-TrkB antibody [EPR17805-146] - Low endotoxin, Azide free (AB223155)

This data was developed using ab187041, the same clone in a different buffer formulation.

Blocking/Dilution buffer : 5% NFDM/TBST.

TrkB is abundantly expressed in the central and peripheral nervous system. The 140 kDa observed MW which is higher than the predicted one is due to the glycosylation modification.

90 kDa corresponds to the truncated form, as observed in the literature (PMID : 24860020).

All lanes:

Western blot - Anti-TrkB antibody [EPR17805-146] (<a href='/en-us/products/primary-antibodies/trkb-antibody-epr17805-146-ab187041'>ab187041</a>) at 1/5000 dilution

Lane 1:

Mouse brain lysate at 10 µg

Lane 2:

Rat brain lysate at 10 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

Predicted band size: 92 kDa

Observed band size: 140 kDa,90 kDa

false

Exposure time: 3min

• WB

Lab

Western blot - Anti-TrkB antibody [EPR17805-146] - Low endotoxin, Azide free (AB223155)

This data was developed using ab187041, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-TrkB antibody [EPR17805-146] (<a href='/en-us/products/primary-antibodies/trkb-antibody-epr17805-146-ab187041'>ab187041</a>) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 20 µg

Lane 2:

Mouse heart tissue lysate at 20 µg

Lane 3:

Mouse lung tissue lysate at 20 µg

Lane 4:

Mouse kidney tissue lysate at 20 µg

Lane 5:

Rat brain tissue lysate at 20 µg

Lane 6:

Rat heart tissue lysate at 20 µg

Lane 7:

Rat lung tissue lysate at 20 µg

Lane 8:

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 1/20000 dilution

Observed band size: 140 kDa,90 kDa

false

Exposure time: 10s