Analysis Note

ControlEGF-stimulated A431 cel lysate, rat brain tissue. Included Positive Antigen Control: Catalog # 12-301, non-stimulated A431 cell lysate. Add 2.5 µL of 2-mercaptoethanol/100 µL of lysate and boil for 5 minutes to reduce the preparation. Load 20 µg of reduced lysate per lane for minigels.

Application

Research Sub CategoryNeurotransmitters & ReceptorsNeurodegenerative Diseases

Research CategoryNeuroscience

Anti-TrkA Antibody detects level of TrkA & has been published & validated for use in IC, IP & WB.

Immunoprecipitation:
4 µg of a previous lot immunoprecipitated TrkA from 500 µg of PC-12 RIPA lysate.

Immunocytochemistry:
10 µg/mL of a previous lot of this antibody showed positive immunostaining for TrkA in PC-12 cells fixed with 4% paraformaldehyde.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General description

TrkA, apparent molecular weight 140 kDa, is a high affinity nerve growth factor (NGF). The Trk proto-oncogene family contains four members, TrkA, TrkB, TrkC, and TrkE, which are variably expressed throughout the central and peripheral nervous systems. TrkA binds to nerve growth factor (NGF) and autophosphorylates on tyrosine residues (Tyr490, Tyr674, Tyr675, Tyr751 and Tyr785) to activate multiple downstream effector proteins. Phosphorylation at Tyr490 is required for Shc association and subsequent activation of the Ras-MAP kinase-signaling cascade, which leads to activation of Elk-1-dependent gene transcription and neurite growth. Phosphorylations at Tyr674 and Tyr675 lie within the catalytic domain of TrkA tyrosine kinase and reflect Trk kinase activity. Additionally, phosphorylation at Tyr751 is required for PI3-kinase association and activation of the Akt signaling cascade.

Immunogen

Rabbits immunized with bacterially expressed purified protein corresponding to the entire extracellular domain of rat TrkA receptor.

Legal Information

UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

Linkage

Replaces: CBL770

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Physical form

Protein A purified

Purified rabbit IgG in buffer containing 0.02 M phosphate buffer, pH 7.6, 0.25 M NaCl, 0.1% sodium azide. Frozen at -20°C.

Format: Purified

Quality

Routinely evaluated by Western Blot on RIPA lysates from human A431 cells, mouse 3T3 or rat PC-12 cell lysates.

Western Blot Analysis:
0.5-2 µg/mL of this lot detected TrkA in RIPA lysates from human A431 cells; a previous lot detected TrkA in mouse 3T3 and rat PC-12 cell lysates.

Specificity

Reactivity with other species has not been confirmed.

Recognizes TrkA, Mr 140 kDa. Does not cross-react with TrkB or TrkC.

Storage and Stability

Stable for 1 year at -20°C from date of receipt.
Handling Recommendations: Upon receipt, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance. Note: Variability in freezer temperatures below -20°C may cause glycerolcontaining solutions to become frozen during storage.

Target description

140 kDa