Product: Phospho-Trk B (Tyr516) Antibody
Catalog: AF3462
Description: Rabbit polyclonal antibody to Phospho-Trk B (Tyr516)
Application: WB IHC IF/ICC
Reactivity: Human, Mouse, Rat
Prediction: Pig, Horse, Sheep, Rabbit, Chicken
Mol.Wt.: 145kDa; 92kD(Calculated).
Uniprot: Q16620
RRID: AB_2834900

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 100ul $280 In stock
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Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Prediction:
Pig(100%), Horse(100%), Sheep(100%), Rabbit(100%), Chicken(100%)
Clonality:
Polyclonal
Specificity:
Phospho-Trk B (Tyr516) Antibody detects endogenous levels of Trk B only when phosphorylated at Tyrosine 516.
RRID:
AB_2834900
Cite Format: Affinity Biosciences Cat# AF3462, RRID:AB_2834900.
Conjugate:
Unconjugated.
Purification:
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

AI848316; BDNF tropomyosine receptor kinase B; BDNF/NT 3 growth factors receptor; BDNF/NT-3 growth factors receptor; Brain derived neurotrophic factor receptor; C030027L06Rik; EC 2.7.10.1; GP145 TrkB; GP145-TrkB; GP145-TrkB/GP95-TrkB; GP95 TrkB; Neurotrophic receptor tyrosine kinase 2; Neurotrophic tyrosine kinase receptor type 2; Neurotrophin receptor tyrosine kinase type 2; NTRK 2; Ntrk2; NTRK2_HUMAN; Obesity, hyperphagia, and developmental delay, included; RATTRKB1; Tkrb; Trk B; Trk-B; TRKB; TrkB tyrosine kinase; TRKB1; Tropomyosin related kinase B; tyrosine kinase receptor B; Tyrosine receptor kinase B;

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Expression:
Q16620 NTRK2_HUMAN:

Isoform TrkB is expressed in the central and peripheral nervous system. In the central nervous system (CNS), expression is observed in the cerebral cortex, hippocampus, thalamus, choroid plexus, granular layer of the cerebellum, brain stem, and spinal cord. In the peripheral nervous system, it is expressed in many cranial ganglia, the ophthalmic nerve, the vestibular system, multiple facial structures, the submaxillary glands, and dorsal root ganglia. Isoform TrkB-T1 is mainly expressed in the brain but also detected in other tissues including pancreas, kidney and heart. Isoform TrkB-T-Shc is predominantly expressed in the brain.

Description:
This gene encodes a member of the neurotrophic tyrosine receptor kinase (NTRK) family. This kinase is a membrane-bound receptor that, upon neurotrophin binding, phosphorylates itself and members of the MAPK pathway. Signalling through this kinase leads to cell differentiation.
Sequence:
MSSWIRWHGPAMARLWGFCWLVVGFWRAAFACPTSCKCSASRIWCSDPSPGIVAFPRLEPNSVDPENITEIFIANQKRLEIINEDDVEAYVGLRNLTIVDSGLKFVAHKAFLKNSNLQHINFTRNKLTSLSRKHFRHLDLSELILVGNPFTCSCDIMWIKTLQEAKSSPDTQDLYCLNESSKNIPLANLQIPNCGLPSANLAAPNLTVEEGKSITLSCSVAGDPVPNMYWDVGNLVSKHMNETSHTQGSLRITNISSDDSGKQISCVAENLVGEDQDSVNLTVHFAPTITFLESPTSDHHWCIPFTVKGNPKPALQWFYNGAILNESKYICTKIHVTNHTEYHGCLQLDNPTHMNNGDYTLIAKNEYGKDEKQISAHFMGWPGIDDGANPNYPDVIYEDYGTAANDIGDTTNRSNEIPSTDVTDKTGREHLSVYAVVVIASVVGFCLLVMLFLLKLARHSKFGMKGPASVISNDDDSASPLHHISNGSNTPSSSEGGPDAVIIGMTKIPVIENPQYFGITNSQLKPDTFVQHIKRHNIVLKRELGEGAFGKVFLAECYNLCPEQDKILVAVKTLKDASDNARKDFHREAELLTNLQHEHIVKFYGVCVEGDPLIMVFEYMKHGDLNKFLRAHGPDAVLMAEGNPPTELTQSQMLHIAQQIAAGMVYLASQHFVHRDLATRNCLVGENLLVKIGDFGMSRDVYSTDYYRVGGHTMLPIRWMPPESIMYRKFTTESDVWSLGVVLWEIFTYGKQPWYQLSNNEVIECITQGRVLQRPRTCPQEVYELMLGCWQREPHMRKNIKGIHTLLQNLAKASPVYLDILG

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
100
Horse
100
Sheep
100
Chicken
100
Rabbit
100
Bovine
0
Dog
0
Xenopus
0
Zebrafish
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - Q16620 As Substrate

Site PTM Type Enzyme
N67 N-Glycosylation
N95 N-Glycosylation
N121 N-Glycosylation
S141 Phosphorylation
T151 Phosphorylation
S153 Phosphorylation
T161 Phosphorylation
N178 N-Glycosylation
N205 N-Glycosylation
N241 N-Glycosylation
T253 Phosphorylation
N254 N-Glycosylation
S256 Phosphorylation
S257 Phosphorylation
S260 Phosphorylation
N280 N-Glycosylation
N338 N-Glycosylation
N412 N-Glycosylation
K461 Acetylation
S479 Phosphorylation
Y490 Phosphorylation
T506 Phosphorylation
Y516 Phosphorylation Q16620 (NTRK2)
Y558 Phosphorylation
T573 Phosphorylation
Y670 Phosphorylation
Y674 Phosphorylation
Y675 Phosphorylation
S698 Phosphorylation
Y702 Phosphorylation
Y706 Phosphorylation
Y707 Phosphorylation
Y785 Phosphorylation
Y817 Phosphorylation

PTMs - Q16620 As Enzyme

Substrate Site Source
O43559 (FRS3) Y417 Uniprot
O43559 (FRS3) Y455 Uniprot
P22001 (KCNA3) Y161 Uniprot
P22001 (KCNA3) Y162 Uniprot
P22001 (KCNA3) Y163 Uniprot
P22001 (KCNA3) Y499 Uniprot
Q00535 (CDK5) Y15 Uniprot
Q13009 (TIAM1) Y829 Uniprot
Q16620 (NTRK2) Y516 Uniprot
Q16620-4 (NTRK2) Y532 Uniprot
Q16620-1 (NTRK2) Y702 Uniprot
Q16620-1 (NTRK2) Y706 Uniprot
Q16620-1 (NTRK2) Y707 Uniprot
Q16620-4 (NTRK2) Y718 Uniprot
Q16620-4 (NTRK2) Y722 Uniprot
Q16620-4 (NTRK2) Y723 Uniprot
Q16620-1 (NTRK2) Y817 Uniprot
Q16620-4 (NTRK2) Y833 Uniprot
Q92529-2 (SHC3) Y218 Uniprot
Q92529-2 (SHC3) Y219 Uniprot
Q92529-2 (SHC3) Y256 Uniprot
Q92529-2 (SHC3) Y257 Uniprot
Q92529-2 (SHC3) Y283 Uniprot
Q92529-2 (SHC3) Y301 Uniprot

Research Backgrounds

Function:

Receptor tyrosine kinase involved in the development and the maturation of the central and the peripheral nervous systems through regulation of neuron survival, proliferation, migration, differentiation, and synapse formation and plasticity (By similarity). Receptor for BDNF/brain-derived neurotrophic factor and NTF4/neurotrophin-4. Alternatively can also bind NTF3/neurotrophin-3 which is less efficient in activating the receptor but regulates neuron survival through NTRK2. Upon ligand-binding, undergoes homodimerization, autophosphorylation and activation. Recruits, phosphorylates and/or activates several downstream effectors including SHC1, FRS2, SH2B1, SH2B2 and PLCG1 that regulate distinct overlapping signaling cascades. Through SHC1, FRS2, SH2B1, SH2B2 activates the GRB2-Ras-MAPK cascade that regulates for instance neuronal differentiation including neurite outgrowth. Through the same effectors controls the Ras-PI3 kinase-AKT1 signaling cascade that mainly regulates growth and survival. Through PLCG1 and the downstream protein kinase C-regulated pathways controls synaptic plasticity. Thereby, plays a role in learning and memory by regulating both short term synaptic function and long-term potentiation. PLCG1 also leads to NF-Kappa-B activation and the transcription of genes involved in cell survival. Hence, it is able to suppress anoikis, the apoptosis resulting from loss of cell-matrix interactions. May also play a role in neutrophin-dependent calcium signaling in glial cells and mediate communication between neurons and glia.

PTMs:

Phosphorylated. Undergoes ligand-mediated autophosphorylation that is required for interaction with SHC1 and PLCG1 and other downstream effectors. Isoform TrkB-T-Shc is not phosphorylated.

Ubiquitinated. Undergoes polyubiquitination upon activation; regulated by NGFR. Ubiquitination regulates the internalization of the receptor (By similarity).

Subcellular Location:

Cell membrane>Single-pass type I membrane protein. Endosome membrane>Single-pass type I membrane protein. Early endosome membrane. Cell projection>Axon. Cell projection>Dendrite. Cytoplasm>Perinuclear region. Cell junction>Synapse>Postsynaptic density.
Note: Internalized to endosomes upon ligand-binding.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Isoform TrkB is expressed in the central and peripheral nervous system. In the central nervous system (CNS), expression is observed in the cerebral cortex, hippocampus, thalamus, choroid plexus, granular layer of the cerebellum, brain stem, and spinal cord. In the peripheral nervous system, it is expressed in many cranial ganglia, the ophthalmic nerve, the vestibular system, multiple facial structures, the submaxillary glands, and dorsal root ganglia. Isoform TrkB-T1 is mainly expressed in the brain but also detected in other tissues including pancreas, kidney and heart. Isoform TrkB-T-Shc is predominantly expressed in the brain.

Subunit Structure:

Exists in a dynamic equilibrium between monomeric (low affinity) and dimeric (high affinity) structures. Interacts (phosphorylated upon activation by BDNF) with SHC1; mediates SHC1 phosphorylation and activation. Interacts (phosphorylated upon activation by BDNF) with PLCG1 and/or PLCG2; mediates PLCG1 phosphorylation and activation. Interacts with SH2B1 and SH2B2. Interacts with NGFR; may regulate the ligand specificity of the receptor (By similarity). Interacts with SORCS2; this interaction is important for normal targeting to post-synaptic densities in response to high-frequency stimulation (By similarity). Interacts (phosphorylated upon ligand-binding) with SH2D1A; regulates NTRK2. Interacts with SQSTM1 and KIDINS220 (By similarity). Interacts (phosphorylated upon ligand-binding) with FRS2; activates the MAPK signaling pathway. Interacts with APPL1 (By similarity). Interacts with MAPK8IP3/JIP3 and KLC1; interaction with KLC1 is mediated by MAPK8IP3/JIP3 (By similarity). Interacts with SORL1; this interaction facilitates NTRK2 trafficking between synaptic plasma membranes, postsynaptic densities and cell soma, hence positively regulates BDNF signaling (By similarity).

Family&Domains:

Belongs to the protein kinase superfamily. Tyr protein kinase family. Insulin receptor subfamily.

Research Fields

· Environmental Information Processing > Signal transduction > MAPK signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > Ras signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > PI3K-Akt signaling pathway.   (View pathway)

· Human Diseases > Substance dependence > Alcoholism.

· Organismal Systems > Nervous system > Neurotrophin signaling pathway.   (View pathway)

References

1). Ding et al. BDNF promotes activation of astrocytes and microglia contributing to neuroinflammation and mechanical allodynia in cyclophosphamide-induced cystitis. Journal of Neuroinflammation 2020 Jan 13;17(1):19 (PubMed: 31931832) [IF=9.3]

Application: WB    Species: rats    Sample: spinal dorsal horn (SDH)

Fig. 1 The expression of BDNF-TrkB signaling in the SDH of CYP-induced cystitis. a Changes of the mechanical threshold in CYP-induced cystitis model. Compared to that in the control group, the mechanical threshold of the cystitis group decreased significantly after the CYP injection and remained low until day 17, and the minimum threshold value was reached on day 12. The expression of b BDNF, c TrkB, and d p-TrkB were evaluated by western blots. Compared to the control group, they were upregulated on days 8, 12, and 17. e Immunofluorescence double staining assay of BDNF and p-TrkB in the SDH. BDNF and p-TrkB (red), NeuN, GFAP, and OX-42( green), co-localization (yellow). BDNF was mainly colocalized in neurons which mainly located in Laminate II to IV. And TrkB receptors expressed in neurons, microglia, and astrocytes. The white dotted lines in picture “BDNF/NeuN” showed the laminate of the SDH according to Rexed and Steiner. Scale bar = 100 μm. All data were calculated as mean ± SEM (n = 5 per group). *p < 0.05, ** p < 0.01, *** p < 0.001 vs. the control group

Application: IF/ICC    Species: rats    Sample: spinal dorsal horn (SDH)

Fig. 1 The expression of BDNF-TrkB signaling in the SDH of CYP-induced cystitis. a Changes of the mechanical threshold in CYP-induced cystitis model. Compared to that in the control group, the mechanical threshold of the cystitis group decreased significantly after the CYP injection and remained low until day 17, and the minimum threshold value was reached on day 12. The expression of b BDNF, c TrkB, and d p-TrkB were evaluated by western blots. Compared to the control group, they were upregulated on days 8, 12, and 17. e Immunofluorescence double staining assay of BDNF and p-TrkB in the SDH. BDNF and p-TrkB (red), NeuN, GFAP, and OX-42( green), co-localization (yellow). BDNF was mainly colocalized in neurons which mainly located in Laminate II to IV. And TrkB receptors expressed in neurons, microglia, and astrocytes. The white dotted lines in picture “BDNF/NeuN” showed the laminate of the SDH according to Rexed and Steiner. Scale bar = 100 μm. All data were calculated as mean ± SEM (n = 5 per group). *p < 0.05, ** p < 0.01, *** p < 0.001 vs. the control group

2). Nanoparticle-mediated intravesical delivery of conditioned medium derived from mesenchymal stem cells for interstitial cystitis/bladder pain syndrome treatment. Applied Materials Today [IF=8.3]

3). Prenatal alcohol exposure enhanced alcohol preference and susceptibility to PTSD in a sex-dependent manner through the synaptic HCN1 channel. Journal of Affective Disorders [IF=6.6]

4). Chen YS et al. Early environmental enrichment for autism spectrum disorder Fmr1 mice models has positive behavioral and molecular effects. EXPERIMENTAL NEUROLOGY 2022 Mar 5;114033. (PubMed: 35259351) [IF=5.3]

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