Custom human CD97 ELISA Kit

Product Features

Product Name           Custom human CD97 ELISA Kit
Assay Type                   Solid Phase Sandwich ELISA
Format                           96-well strip plate
Assay Length              4.5 hours
Sample Volume         Serum ,Plasma, cell culture media , 100ul
Detection Method    Colorimetric
Sensitivity                   This has not yet been determined
Assay Range                This has not yet been determined
Reactivity                     Human
Cross-Reactivity       This ELISA antibody pair only recognizes human CD97
Gene Names:              ADGRE5, CD97, TM7LN1
Gene ID                         976
Swissprot ID               P48960

Kit Components

• Pre-Coated 96-well Strip Microplate
• Wash Buffer
• Stop Solution
• Assay Diluent
• Standards
• Biotinylated Detection Antibody
• Streptavidin-Conjugated HRP
• TMB One-Step Substrate

Protocol Outline

1. Prepare all reagents, samples and standards as instructed in the manual.
2. Add 100 µl of standard or sample to each well.
3. Incubate 2.5 h at RT or O/N at 4°C.
4. Wash and add 100 µl of prepared biotin antibody to each well.
5. Incubate 1 h at RT.
6. Wash and add 100 µl of prepared Streptavidin solution to each well.
7. Incubate 45 min at RT.
8. Wash and add 100 µl of TMB One-Step Substrate Reagent to each well.
9. Incubate 30 min at RT.
10. Add 50 µl of Stop Solution to each well.
11. Read at 450 nm/570 nm.

Background

CD97 is widely expressed on, among others, hematopoietic stem and progenitor cells, immune cells, epithelial cells, muscle cells as well as their malignant counterparts. In the case of CD97 the N-terminal domains consist of alternatively spliced epidermal growth factor (EGF)-like domains. Transgenic expression of a CD97 in mice enhanced levels of nonphosphorylated membrane-bound β-catenin and phosphorylated Akt. Furthermore, ectopic CD97 expression facilitated RhoA activation through binding of Gα12/13 as well as induced Ki67 expression and phosphorylated ERK and Akt through enhancing lysophosphatidic acid receptor 1 (LPAR1) signaling. Outside the immune system, CD97 is likely involved in cell–cell interactions. CD97 in colonic enterocytes strengthens E-cadherin-based adherens junctions to maintain lateral cell-cell contacts and regulates the localization and degradation of β-catenin through glycogen synthase kinase-3β (GSK-3β) and Akt signaling.

Storage

The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.

Regulatory Status      

Custom human CD97 ELISA Kit is for Research Use Only.

• Manual
• Assay procedure
• MSDS