The relationship between the protein concentration of lysates from untreated and H202-treated Jurkat cells and the absorbance at 450 nm using the PathScan® Phospho-Lck (Tyr505) Sandwich ELISA Antibody Pair is shown. Jurkat cells were treated with H202 (20 mM) for 3 minutes at 37ºC and then lysed.
Capture and detection antibodies are stored at 4°C. HRP-linked secondary reagent is stored at -20°C.
Cell Signaling Technology's PathScan® Phospho-Lck (Tyr505) Sandwich ELISA Antibody Pair is being offered as an economical alternative to our PathScan® Phospho-Lck (Tyr505) Sandwich ELISA Kit #7941. Capture and detection antibodies (100X stocks) and an HRP-conjugated secondary antibody (1000X stock) are supplied. Sufficient reagents are supplied for 4 x 96 well ELISAs. The phospho-Lck (Tyr505) rabbit capture antibody is coated onto a 96 well microplate overnight in PBS. After blocking, cell lysates are added followed by a total Lck mouse detection antibody and anti-mouse IgG, HRP-linked antibody. HRP substrate (TMB) is then added for color development. The magnitude of the absorbance for this developed color is proportional to the quantity of phospho-Lck (Tyr505) protein.
For Antibody Pair specificity and sensitivity, please refer to the corresponding PathScan® Sandwich ELISA Kit. Note: This antibody pair detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
Lck belongs to the Src-like non-receptor tyrosine kinase family with the typical Src family kinase structure: a unique amino terminal domain (Src homology 4 domain, SH4) followed by an SH3 domain, an SH2 domain, a kinase domain (SH1), and a carboxy-terminal negative regulatory domain (1). Lck activity is controlled by the interactions of SH2 and SH3 domains as well as tyrosine phosphorylation status of the activation loop (2,3). Lck is recruited to the T cell receptor (TCR) complex upon stimulation and activates downstream tyrosine kinases to initiate T cell signaling (4). Lck is also found to be involved in the regulation of mitochondrial apoptosis pathways and may be responsible for some anticancer drug induced apoptosis (5,6).
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PathScan is a trademark of Cell Signaling Technology, Inc.