Klotho beta ELISA Kit

Catalog No. ABIN5510659

This Colorimetric ELISA kit is designed for the quantitative measurement of Human Klotho beta.

Quick Overview for Klotho beta ELISA Kit (ABIN5510659)

Target: Klotho beta (KLB)

Binding Specificity: AA 53-997

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 156 pg/mL - 10000 pg/mL

Application: ELISA

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

Product Details Klotho beta ELISA Kit

Minimum Detection Limit: 156 pg/mL

Purpose: Sandwich Human Klotho beta ELISA Kit to quantitate Human KLB in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA).

Analytical Method: Quantitative

Specificity: Expression system for standard: NS0
Immunogen sequence: F53-L997

Capture antibody and Detection antibody: monoclonal antibody from mouse, 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%): 6.1%, 6.6%, 5.4%
Inter-Assay Precision (CV%): 4.3%, 4.8%, 7.5%

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 Klotho beta

Target: Klotho beta (KLB)

Alternative Name: KLB

Background:

Background: Klotho beta, a divergent structural member of the glycosidase I superfamily, is expressed primarily in the liver and pancreas, with lower expression in adipose tissue. It is mapped to chromosome 4. The Klotho beta mediated interaction of human FGF19 (mouse FGF15) with FGF Receptor 4 in the liver negatively regulates bile acid synthesis by controlling the secretion of two key bile acid synthase genes, cholesterol 7-alpha hydroxylase (Cyp7a1) and sterol 12-alpha hydroxylase (Cyp8b1). Klotho beta is also an essential cofactor for FGF21 activity and the Klotho gene family may have evolved to confer tissue-specific bioactivity on FGF19 subfamily members.

Gene Full Name: klotho beta

Gene ID: 152831

UniProt: Q86Z14

Pathways: Fc-epsilon Receptor Signaling Pathway, EGFR Signaling Pathway, Neurotrophin Signaling Pathway, Growth Factor Binding