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Mouse monoclonal antibody to Tyrosine Kinase Receptor A (TrkA, 604-615), intracellular domain [Clone BS292]: IgG

$297.00USD


Catalogue No. M-1719-100
Description TrkA is a member of the neurotrophic tyrosine kinase receptor family. It is a membrane-bound receptor that upon neurotrophin binding, phosphorylates itself and members of the MAPK pathway. TrkA is required for high-affinity binding to nerve growth factor (NGF), neurotrophin-3 and neurotrophin-4/5 but not brain-derived neurotrophic factor (BDNF). TrkA leads to cell differentiations and may play a role in specifying sensory neuron subtypes. It has a crucial role in the development and function of the nociceptive reception system as well as establishment of thermal regulation via sweating. SUBUNIT: Exists in a dynamic equilibrium between monomeric (low affinity) and dimeric (high affinity) structures. SUBCELLULAR LOCATION: Cell membrane; single-pass type I membrane protein. Endocytosed to the endosomes upon treatment of cells with NGF. ALTERNATIVE PRODUCTS: 2 named isoforms produced by alternative splicing. Both isoforms have similar biological properties. TISSUE SPECIFICITY: Isoform TrkA-II is primarily expressed in neuronal cells. Isoform TrkA-I is found in non-neuronal tissues. Mutations in TrkA have been associated with congenital insensitivity to pain, anhidrosis, self-mutalating behaviour, mental retardation and cancer (Reference: www.uniprot.com).
Batch No. Please refer to the product label.
Unit size 100 g
Antigen A synthetic peptide from the intracellular cytoplasmic domain of human TrkA (C-HGPDAKLLAGGE, aa: 604-615) has been used as the immunogen.
Sequence HGPDAKLLAGGE
Antigen Location Antigen's peptide sequence is located within the cytoplasmic domain consisting of amno acids 440 796.
Antigen Length 12 amino acids
Antibody Type Mouse monoclonal IgG
Isotype IgG3, k-light chain
Clone BS292
Other Names Tropomyosin-related kinase receptor; High affinity nerve growth factor receptor; Neurotrophic tyrosine kinase receptor type 1; TRK1 transforming tyrosine kinase protein; p140-TrkA; Trk-A; NTRK1; TRK
Accession Uniprot: P04629; NTRK1_HUMAN
Produced in Mouse
Molecular Weight TrkA 140 kDa, on SDS-PAGE gels, with isoforms at 110 kDa and internal cleavage products at 41 kDa
Purity Protein G purified mouse immunoglobulin
Applications WB: Western blotting: 1-3 g/mL, antibody is suitable for reduced (with DTT or β-mercaptoethanol) and non-reduced samples. Denatured but not reduced samples provides cleaner blot signals as demonstrated in our photographs.

Immunocytochemistry: 1-2 g/mL. Antibody works on 4% formaldehyde-fixed cells. Note that cells require a permeabilization step, because the antibody detects a cytoplasmic epitope of TrkA.

Other applications not yet tested. Biosensis recommends optimal dilutions/concentrations should be determined by the end user.
Specificity TrkA. Does not cross-react with TrkB or TrkC.
Species Against Human
Antibody Against Human
Cross-reactivity Reacts with human TrkA. Known to cross-react with TrkA from rat and mouse. Expected to cross-react with other mammalian species based on peptide antigen sequence similarity.
Form Lyophilized from PBS, pH 7.4, containing 3% trehalose without preservatives.
Reconstitution Spin vial briefly before opening. Reconstitute in 100 l of sterile water. Centrifuge to remove any insoluble material. Final buffer contains no preservatives.
Storage After reconstitution divide into aliquots and store at -20C for a higher stability. Antibody contains no preservatives. Store at 4C with an appropriate antibacterial agent. Use sterile methods. Highest purity Glycerol (1:1) may be added for an additional stability when stored at refrigerated or freezing temperatures. Avoid repetitive freeze/thaw cycles.
Expiry Date 12 months after purchase if unopened.
General References 1. Cabrera N. et al., TrkA receptor ectodomain cleavage generates a tyrosine-phosphorylated cell-associated frangment. J Cell Biol, 1996, Vol.132(3), pp.427-436.
2. Rossi F.M. et al., Expression of the nerve growth factor receptors TrkA and p75NTR in the visual cortex of the rat: development and regulation by the cholinergic input. J Neurosci, Februar 1, 2002, 22(3):912-919.
Images (click to zoom)
Mouse monoclonal antibody to Tyrosine Kinase Receptor A (TrkA, 604-615), intracellular domain [Clone BS292]: IgG Western blot of TrkA in rat PC12 cell lysates (64 ug/lane): SDS-PAGE: denatured, non-reduced; Transfer: Tris-Glycine buffer; Membrane: nitrocellulose (0.45 um); Blocking: 5% skim milk in TBST, 1 hour at RT; Primary antibody: overnight at 4°C (3 ug/mL); Secondary antibody: anti-mouse-HRP (1/6000) 2 hours at RT; Detection: Chemiluminiscence. M-1719-100 detects TrkA full-length protein at approximately 140 kDa. Predicted MW of rat TrkA based on amino acid sequence: 85 kDa. The observed MW differs due to post-translational modification, mainly glycosylation.
Mouse monoclonal antibody to Tyrosine Kinase Receptor A (TrkA, 604-615), intracellular domain [Clone BS292]: IgG Western blot of TrkA in rat PC12 cell lysates (80 μg/lane). M-1719-100 detects an isoform of glycosylated TrkA at 110 kDa and an intracellular cleavage product at 41 kDa (see Cabrera et al., 1995; Rossi et al., 2002). These bands are TrkA-specific, because detection by M-1719-100 is abolished after pre-incubation with blocking peptide. SDS-PAGE: denatured, non-reduced; Transfer: Tris-Glycine buffer; Membrane: nitrocellulose (0.45 μm); Blocking: 5% skim milk in TBST, 1 hour at RT; Primary antibody: overnight at 4C (3 μg/mL); Secondary antibody: anti-mouse-HRP (1/6000) 2 hours at RT; Detection: Chemiluminiscence.
Mouse monoclonal antibody to Tyrosine Kinase Receptor A (TrkA, 604-615), intracellular domain [Clone BS292]: IgG Western blot of TrkA in rat PC12 cell lysates (30 μg/lane). M-1719-100 detects TrkA full-length protein at ≈ 140 kD, an isoform of glycosylated TrkA at 110 kDa and a intracellular cleavage product at 41 kDa (see Cabrera et al., 1995; Rossi et al., 2002). SDS-PAGE: denatured, reduced and non-reduced; Transfer: Tris-Glycine buffer; Membrane: nitrocellulose (0.45 μm); Blocking: 5% skim milk in TBST, 2 hours at RT; Primary antibody: overnight at 4C (3 μg/mL); Secondary antibody: anti-mouse-HRP (1/6000) 2 hours at RT; Detection: Chemiluminiscence.
Mouse monoclonal antibody to Tyrosine Kinase Receptor A (TrkA, 604-615), intracellular domain [Clone BS292]: IgG Immunofluorescence analysis of TrkA expression in human SH-SY5Y cells. Fixed (4% formaldehyde), permeabilized, and blocked (10% normal horse serum, 0.1% Triton X100) SH-SY5Y cells were incubated with TrkA antibody M-1719-100 (2 μg/mL, green) for 1 hour. Primary antibody binding was visualized with a secondary donkey anti-mouse-CF488A antibody (4 μg/mL, 1 hour incubation). Cell nuclei were stained with Hoechst dye (blue). Magnification: 100x.
Mouse monoclonal antibody to Tyrosine Kinase Receptor A (TrkA, 604-615), intracellular domain [Clone BS292]: IgG Immunofluorescence analysis of TrkA expression in human DU145 prostate cancer cells. Fixed (4% formaldehyde), permeabilized, and blocked (10% normal horse serum, 0.1% Triton X100) DU145 cells were incubated with TrkA antibody M-1719-100 (2 μg/mL, green) for 1 hour. Primary antibody binding was visualized with a secondary donkey anti-mouse-CF488A antibody (4 μg/mL, 1 hour incubation). Cell nuclei were stained with Hoechst dye (blue). Magnification: 100x.
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