| Field | Specification |
|---|---|
| Mfr No | |
| Alternative Names | LAC; LPH; LPH1; lactase-glycosylceramidase|lactase-phlorizin hydrolase|lactase-phlorizin hydrolase-1 |
| Product Type | |
| Sensitivity | |
| UniProt # |
Scientific Background
v-CRK avian sarcoma virus CT10-homolog-like contains one SH2 domain and two SH3 domains. CRKL has been shown to activate the RAS and JUN kinase signaling pathways and transform fibroblasts in a RAS-dependent fashion. It is a substrate of the BCR-ABL tyrosine kinase and plays a role in fibroblast transformation by BCR-ABL. In addition, CRKL has oncogenic potential.CrkL together with Crk participates in the Reelin signaling cascade downstream of DAB1.CRKL becomes phosphorylated when overexpressed, activates Ras-dependent and JNK pathways, and transforms fibroblasts. The authors also found CRKL t.
Assay Principle
This assay employs a two-site sandwich ELISA to quantitate LCT in samples. An antibody specific for LCT has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyLCT present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for LCT is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of LCT bound in the initial step. The color development is stopped and the intensity of the color is measured.
Performance Specifications
| Sensitivity | 0.058 ng/mL |
|---|---|
| Detection Range | 0.156-10 ng/mL |
| Total Assay Time | 3.5–5 hours |
| Compatible Sample Types | serum, plasma, cell culture supernatant, tissue homogenate |
| Species Reactivity | Human |
| Detection Method | Colorimetric (TMB/HRP) |
| Storage | 4°C (short-term); -20°C (long-term) |
✓ Research-Grade Validation
Safety & Regulatory
Handle reagents in accordance with institutional biosafety guidelines. Refer to the Safety Data Sheet (SDS) for complete hazard and handling information. Contains components that may require special disposal procedures per local regulations.
This kit is validated for use with serum, plasma, cell culture supernatant, tissue homogenate. For unlisted matrices (e.g., tissue lysate, urine), perform a spike-and-recovery experiment to confirm assay performance before generating reportable data. Sample dilution in the kit's provided diluent is recommended to minimize matrix interference.
The minimum detectable concentration (sensitivity) of this kit is 0.058 ng/mL. Values below this threshold should be reported as below the limit of detection (<LOD) and should not be extrapolated from the standard curve.
The total assay time from sample addition to absorbance reading is approximately 3.5–5 hours, including all incubation, wash, and substrate steps. Hands-on time is typically 1–2 hours; most steps involve passive plate incubation. Plan the assay as a single uninterrupted session for best results.
Standard components of this Sandwich ELISA Kit typically include: pre-coated microplate (96-well strip format), lyophilized or liquid recombinant LCT standard, detection antibody, streptavidin-HRP conjugate, TMB substrate, stop solution, wash buffer concentrate, and sample/standard diluent. Refer to the kit insert or datasheet for the exact component list and storage requirements.
This kit uses colorimetric (TMB/HRP) detection and requires a standard microplate absorbance reader capable of measuring at 450 nm. A reference wavelength of 570 nm or 630 nm is recommended to reduce background. No specialized fluorescence or luminescence reader is needed. Ensure the instrument is calibrated and the plate is clean and free of condensation before reading.
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