MaxLight™550 is a new Yellow-Green photostable dye conjugate comparable to Alexa Fluor™546, 555, DyLight™549 , Cy3™, TRITC and offers better labeling efficiency, brighter imaging and increased immunodetection. Absorbance (550nm); Emission (575nm); Extinction Coefficient 150,000.
TrkA has an apparent molecular weight 140kD and is a high affinity nerve growth factor (NGF).
Applications
Suitable for use in Western Blot, Immunoprecipitation, Immunohistochemistry and Immunocytochemistry. Other applications have not been tested.
Recommended Dilutions
Western Blot: 0.5-2.0ug/ml detected TrkA in RIPA lysates from human A431 cells, TrkA in mouse 3T3 and rat PC-12 cell lysates. A431 cell lysate was resolved by electrophoresis, transferred to nitrocellulose and probed with T8663 (2ug/ml). Proteins were visualized using a Gt x Rb IgG (HRP) and a chemiluminescence detection system. Immunoprecipitation: 4ug immunoprecipitates TrkA from 500ug of PC-12 RIPA lysate. Immunohistochemistry (Paraffin): 1:200 detected TrkA in neurons of the hippocampal region and adrenal gland’s medulla. Immunocytochemistry: 10ug/ml showed positive staining for TrkA in PC-12 cells fixed with 4% paraformaldehyde. Optimal dilutions to be determined by the researcher.
Recommended Secondary Antibodies
I1904-39: IgG, X-Adsorbed (HRP) Pab Gt x Rb I1904-40A: IgG, H&L, X-Adsorbed (HRP) Pab Gt x Rb
Included Positive Antigen Control
A0001-50: Non-stimulated A431 cell lysate. Add 2.5ul of 2-mercaptoethanol/100ul of lysate and boil for 5 minutes to reduce the preparation. Load 20ug of reduced lysate per lane for minigels.
Storage and Stability
Store product at 4°C in the dark. DO NOT FREEZE! Stable at 4°C for 12 months after receipt as an undiluted liquid. Dilute required amount only prior to immediate use. Further dilutions can be made in assay buffer. Caution: MaxLight™550 conjugates are sensitive to light. For maximum recovery of product, centrifuge the original vial prior to removing the cap.
Note: Applications are based on unconjugated antibody.
Immunogen
Bacterially expressed purified protein corresponding to the entire extracellular domain of rat TrkA receptor.
Form
Supplied as a liquid in PBS, pH 7.2. No preservative added. Labeled with MaxLight™550.
Purity
Purified by Protein A affinity chromatography.
Specificity
Recognizes rat TrkA, at ~140kD. Does not crossreact with TrkB or TrkC. Species Crossreactivity: human and mouse