LY9 ELISA Kit

Catalog No. ABIN8055608

Mouse LY9 ELISA Kit, Colorimetric assay for quantification of Mouse LY9.

Quick Overview for LY9 ELISA Kit (ABIN8055608)

Target: LY9 (CD229) (Lymphocyte Antigen 9 (CD229))

Binding Specificity: AA 48-453

Reactivity: Mouse

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 93.7 pg/mL - 6000 pg/mL

Application: ELISA

Sample Type: Cell Culture Supernatant, Plasma (EDTA), Plasma (citrate), Plasma (heparin), Serum

Product Details LY9 ELISA Kit

Minimum Detection Limit: 93.7 pg/mL

Purpose: Sandwich Mouse Ly9/SLAMF3/CD229 ELISA Kit to quantitate Mouse Ly9 in cell culture supernatants, serum and plasma (heparin, EDTA, citrate).

Analytical Method: Quantitative

Specificity: Expression system for standard: NS0
Immunogen sequence: K48-R453

Capture antibody and Detection antibody: a monoclonal antibody from rat|a biotinylated polyclonal antibody from goat

Cross-Reactivity (Details): There is no detectable cross-reactivity with other relevant proteins.

Sensitivity: < 50 pg/mL

Components:

1. Pre-coated 96-well strip microplate
2. Standard
3. Biotinylated antibody (100x)
4. Avidin-Biotin-Peroxidase Complex (100x)
5. Sample Diluent
6. Antibody Diluent
7. Avidin-Biotin-Peroxidase Diluent
8. Color Developing Reagent (TMB)
9. Stop Solution
10. Wash Buffer (25x)
11. Adhesive plate sealers

Material not included: Microplate Reader capable of reading absorbance at 450nm. Incubator. Automated plate washer (optional). Pipettes and pipette tips capable of precisely dispensing 0.5 μL through 1 mL volumes of aqueous solutions. Multichannel pipettes are recommended for large amount of samples. Deionized or distilled water. 500 mL graduated cylinders. Test tubes for dilution.

Application Details

Application Notes: Before using Kit, spin tubes and bring down all components to bottom of tube. Duplicate well assay was recommended for both standard and sample testing.

Sample Volume: 100 μL

Plate: Pre-coated

Protocol: ELISA Kit is based on standard sandwich enzyme-linked immune-sorbent assay technology. An antibody has been precoated onto 96-well plates. Standards and test samples are added to the wells, a biotinylated detection antibody specific for target is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the target amount in sample captured in plate.

Assay Precision: Intra-Assay Precision (CV%): 4.1%, 4.3%, 4.6%
Inter-Assay Precision (CV%): 5.8%, 6.9%, 6.4%

Restrictions: For Research Use only

Handling

Handling Advice: Avoid multiple freeze-thaw cycles.

Storage: 4 °C,-20 °C

Storage Comment: Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)

Expiry Date: 12 months

Target Details for LY9

Target: LY9 (CD229) (Lymphocyte Antigen 9 (CD229))

Alternative Name: Ly9

Background:

Background: T-lymphocyte surface antigen Ly-9 is a protein that in humans is encoded by the LY9 gene. This gene is mapped to 1 H3, 1 79.54 cM. Mature human CD229 consists of a 407 amino acid (aa) extracellular domain (ECD) with two Ig like V-set and two Ig like truncated C2-set domains. It also shows a 22 aa transmembrane segment, and a 179 aa cytoplasmic domain with two immunoreceptor tyrosine-based switch motifs ITSMs. The ECD of human CD229 shares 57 % 59 % aa sequence identity with mouse and rat CD229. Within the first two Ig like domains that are common to all SLAM proteins, human CD229 shares 24 % 39 % aa sequence identity with human 2B4, BLAME, CD2F-10, CD84, CRACC, NTB-A, and SLAM. Alternate splicing generates two additional isoforms that lack the juxtamembrane Ig like domain or short cytoplasmic region. CD229 is expressed on T and B cells, thymocytes, and more weakly on NK cells. Homophilic binding between CD229 Molecules is mediated by the N-terminal Ig like domain. Human and mouse CD229 exhibit cross-species binding. Antigen stimulation of lymphocytes induces CD229 clustering to sites of T cell - B cell contact. Two tyrosines in the cytoplasmic domain are required for association of CD229 with the adaptor proteins AP-2 (μ2 chain) and GRB-2 and both are required for CD229 internalization.

Gene Full Name: lymphocyte antigen 9

Gene ID: 17085

UniProt: Q01965

Pathways: BCR Signaling