Anti-TrkA (phospho Y674)+TrkB (phospho Y706)+TrkC (phospho Y709) antibody [EPR22298-67]

• WB

Unknown

Western blot - Anti-TrkA (phospho Y674)+TrkB (phospho Y706)+TrkC (phospho Y709) antibody [EPR22298-67] (AB229908)

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

Observed MW : 140 kDA

All lanes:

Western blot - Anti-TrkA (phospho Y674)+TrkB (phospho Y706)+TrkC (phospho Y709) antibody [EPR22298-67] (ab229908) at 1/1000 dilution

Lane 1:

Human brain at 20 µg

Lane 2:

Human Hippocampus at 20 µg

Lane 3:

Human cerebellum at 20 µg

Lane 4:

Human spinal cord at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 92 kDa

false

Exposure time: 5s

• WB

Lab

Western blot - Anti-TrkA (phospho Y674)+TrkB (phospho Y706)+TrkC (phospho Y709) antibody [EPR22298-67] (AB229908)

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

Lanes 1 - 4:

Western blot - Anti-TrkA (phospho Y674)+TrkB (phospho Y706)+TrkC (phospho Y709) antibody [EPR22298-67] (ab229908) at 1/1000 dilution

Lanes 1 - 4:

Western blot - Anti-TrkA (pY674) + TrkB (pY706) + TrkC (pY709) antibody [EPR22298-67] - BSA and Azide free (<a href='/en-us/products/primary-antibodies/trka-phospho-y674trkb-phospho-y706trkc-phospho-y709-antibody-epr22298-67-bsa-and-azide-free-ab264070'>ab264070</a>) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a His-tag, whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a human TrkA expression vector containing a His-tag, whole cell lysate at 20 µg

Lane 3:

293T cells transfected with a human TrkB expression vector containing a His-tag, whole cell lysate at 20 µg

Lane 4:

293T cells transfected with a human TrkC expression vector containing a His-tag, whole cell 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 1/100000 dilution

Observed band size: 140 kDa

false

Exposure time: 15s

• WB

Lab

Western blot - Anti-TrkA (phospho Y674)+TrkB (phospho Y706)+TrkC (phospho Y709) antibody [EPR22298-67] (AB229908)

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 : 24959744). Besides, TrkB auto-phosphorylation was observed when TrkB was over-expressed in HEK293 cells, which has been reported in the literature (PMID : 25632160).

Exposure time : 7 seconds

All lanes:

Western blot - Anti-TrkA (phospho Y674)+TrkB (phospho Y706)+TrkC (phospho Y709) antibody [EPR22298-67] (ab229908) at 1/5000 dilution

Lane 1:

HEK-293T (human embryonic kidney) transfected with an empty vector (vector control), containing a myc-His-tag®, whole cell lysate at 10 µg

Lane 2:

HEK-293T transfected with human TrkB (WT) expression vector containing a myc-His-tag®, whole cell lysate (Untreated membrane) at 10 µg

Lane 3:

HEK-293T transfected with human TrkB (WT) expression vector containing a myc-His-tag®, whole cell lysate (Phosphatase treated membrane) 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 1/50000 dilution

Predicted band size: 92 kDa

Observed band size: 140 kDa

false

• WB

Unknown

Western blot - Anti-TrkA (phospho Y674)+TrkB (phospho Y706)+TrkC (phospho Y709) antibody [EPR22298-67] (AB229908)

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

Observed MW : 140 kDA

All lanes:

Western blot - Anti-TrkA (phospho Y674)+TrkB (phospho Y706)+TrkC (phospho Y709) antibody [EPR22298-67] (ab229908) at 1/1000 dilution

Lane 1:

Mouse brain lysates at 20 µg

Lane 2:

Mouse hippocampus at 20 µg

Lane 3:

Mouse P7 thalamus at 20 µg

Lane 4:

Mouse cerebellum at 20 µg

Lane 5:

Rat brain at 20 µg

Lane 6:

Rat hippocampus at 20 µg

Lane 7:

Rat P7 thalamus at 20 µg

Lane 8:

Rat cerebellum at 20 µg

Lane 9:

Mouse P5 cerebral cortex at 20 µg

Lane 10:

Mouse P3 retina at 20 µg

Lane 11:

Mouse P20 retina at 20 µg

Lane 12:

Rat spinal cord 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 1/20000 dilution

Predicted band size: 92 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-TrkA (phospho Y674)+TrkB (phospho Y706)+TrkC (phospho Y709) antibody [EPR22298-67] (AB229908)

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

The phosphorylation sites are conserved between TrkA and TrkB : Tyr674/675 of TrkA corresponds to Tyr706/707 in TrkB. Therefore, the TrkB (phospho T705) antibody can detect the phosphorylation of TrkA T674/675 in PC-12 cells treated with NGF. The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 23951412).

Exposure time : 3 minutes

All lanes:

Western blot - Anti-TrkA (phospho Y674)+TrkB (phospho Y706)+TrkC (phospho Y709) antibody [EPR22298-67] (ab229908) at 1/1000 dilution

Lane 1:

Untreated PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate at 10 µg

Lane 2:

PC-12 treated with 100ng/ml NGF (<a href='/en-us/products/proteins-peptides/recombinant-human-ngf-protein-active-ab9796'>ab9796</a>) for 5 minutes 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: 92 kDa

Observed band size: 140 kDa

false

• WB

Lab

Western blot - Anti-TrkA (phospho Y674)+TrkB (phospho Y706)+TrkC (phospho Y709) antibody [EPR22298-67] (AB229908)

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 : 24959744). Besides, TrkB auto-phosphorylation was observed when TrkB was over-expressed in HEK293 cells, which has been reported in the literature (PMID : 25632160).

Exposure time : 48 seconds

All lanes:

Western blot - Anti-TrkA (phospho Y674)+TrkB (phospho Y706)+TrkC (phospho Y709) antibody [EPR22298-67] (ab229908) at 1/5000 dilution

Lane 1:

HEK-293T (human embryonic kidney) transfected with an empty vector (vector control), containing a myc-His-tag®, whole cell lysate at 10 µg

Lane 2:

HEK-293T transfected with mouse TrkB (WT) expression vector containing a myc-His-tag®, starved for 6 hours, whole cell lysate at 10 µg

Lane 3:

HEK-293T transfected with mouse TrkB (WT) expression vector containing a myc-His-tag®, starved for 6 hours, then treated with 50ng/ml BDNF (<a href='/en-us/products/proteins-peptides/recombinant-human-murine-rat-bdnf-protein-active-ab9794'>ab9794</a>) for 10 minutes, whole cell lysate (Untreated membrane) at 10 µg

Lane 4:

HEK-293T transfected with mouse TrkB (WT) expression vector containing a myc-His-tag®, starved for 6 hours, then treated with 50ng/ml BDNF (<a href='/en-us/products/proteins-peptides/recombinant-human-murine-rat-bdnf-protein-active-ab9794'>ab9794</a>) for 10 minutes, whole cell lysate (Phosphatase treated membrane) at 10 µg

Lane 5:

HEK-293T transfected with mouse TrkB (mutant Y705A) expression vector containing a myc-His-tag®, whole cell 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 1/50000 dilution

Predicted band size: 92 kDa

Observed band size: 140 kDa

false

• WB

Lab

Western blot - Anti-TrkA (phospho Y674)+TrkB (phospho Y706)+TrkC (phospho Y709) antibody [EPR22298-67] (AB229908)

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

ab181602 was used as a GAPDH loading control at 1/200000 dilution.

All lanes:

Western blot - Anti-TrkA (phospho Y674)+TrkB (phospho Y706)+TrkC (phospho Y709) antibody [EPR22298-67] (ab229908) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a His-tag, whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a human TrkA expression vector containing a His-tag, whole cell lysate at 20 µg

Lane 3:

293T cells transfected with a human TrkB expression vector containing a His-tag, whole cell lysate at 20 µg

Lane 4:

293T cells transfected with a human TrkC expression vector containing a His-tag, whole cell 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 1/100000 dilution

Observed band size: 95 kDa,140 kDa

false

Exposure time: 15s

• Dot

Unknown

Dot Blot - Anti-TrkA (phospho Y674)+TrkB (phospho Y706)+TrkC (phospho Y709) antibody [EPR22298-67] (AB229908)

Dot blot analysis using ab229908 at 1/1000 dilution, followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100,000 dilution.

Lane 1 : TrkB (phospho T705) peptide (aa697-708)

Lane 2 : TrkB (phospho T705) peptide (aa702-712)

Lane 3 : TrkB non-phospho peptide (aa697-712)

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

• Dot

Lab

Dot Blot - Anti-TrkA (phospho Y674)+TrkB (phospho Y706)+TrkC (phospho Y709) antibody [EPR22298-67] (AB229908)

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

All lanes:

Dot Blot - Anti-TrkA (phospho Y674)+TrkB (phospho Y706)+TrkC (phospho Y709) antibody [EPR22298-67] (ab229908) at 1/1000 dilution

Lane 1:

Human TrkA (phospho Y674) peptide

Lane 2:

Human TrkA (phospho Y675) peptide

Lane 3:

Human TrkA (phospho Y674/675) peptide1

Lane 4:

Human TrkA (phospho Y674/675) peptide2

Lane 5:

Human TrkA control peptide for Lane 1-4

Lane 6:

Human TrkB (phospho Y706) peptide

Lane 7:

Human TrkB (phospho Y707) peptide

Lane 8:

Human TrkB (phospho Y706/707) peptide

Lane 9:

Human TrkB control peptide for Lane 6-8

Lane 10:

Human TrkC (phospho Y709) peptide

Lane 11:

Human TrkC (phospho Y710) peptide

Lane 12:

Human TrkC (phospho Y709/710) peptide

Lane 13:

Human TrkC control peptide for Lane 10-12

Secondary

All lanes:

Dot 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

false

Exposure time: 180s