Mouse Klotho (KI) ELISA Kit (Part mKI-ELISA 96T)

Mouse Klotho (KI) ELISA Kit

Introduction

Membrane bound Klotho is 130kDa type-I membrane protein that is related to β-glucuronidases. Shedding of the ectodomain results in the generation of 70kDa soluble alpha-klotho fragment detected by this ELISA kit. Reduced production of this protein has been observed in patients with chronic renal failure (CRF), and this may be one of the factors underlying the degenerative processes (e.g., arteriosclerosis, osteoporosis, and skin atrophy) seen in CRF. Also, mutations within this protein have been associated with ageing, bone loss and alcohol consumption. Transgenic mice that overexpress Klotho live longer than wild-type mice. Klotho has been characterized as modulating amyloid beta pathogenesis in Alzheimers disease.

Protocol

Specifications

Product name

Mouse Kl(Klotho) ELISA Kit

Catalogue No.

mKI-ELISA

Alias

Kl ELISA Kit

Detection method

Sandwich ELISA, Double Antibody

Application

KI ELISA kit allows for the in vitro quantitative determination of Kl/KL/klotho/KLOTHO concentrations in plasma, tissue homogenates and other biological fluids.

Size

96T

Range

15.6-1000pg/ml

Sensitivity

< 9.375pg/ml

Standard curve

Mouse Kl(Klotho) ELISA Kit

Storage

4 ℃ for 6 months

Recovery

Matrices listed below were spiked with certain level of Kl/KL/klotho/KLOTHO and the recovery rates were calculated by comparing the measured value to the expected amount of Kl/KL/klotho/KLOTHO in samples.

Matrix	Recovery range(%)	Average(%)
serum(n=5)	85-102	97
EDTA plasma(n=5)	91-103	97
heparin plasma(n=5)	85-97	90

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Kl/KL/klotho/KLOTHO and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample	1:2	1:4	1:8	1:16
serum(n=5)	100-104%	93-100%	91-105%	86-104%
EDTA plasma(n=5)	84-101%	88-98%	83-101%	87-101%
heparin plasma(n=5)	87-100%	80-97%	86-100%	80-100%

CV(%)

Intra-Assay: CV<8%
Inter-Assay: CV<10%

Kit Components Item Specifications (48T/96T)

Storage Micro ELISA Plate (Dismountable) 8×6 or 8×12 4°C/-20°C
Lyophilized Standard 1 vial or 2 vial 4°C/-20°C Sample
Standard dilution buffer 10ml/20ml 4°C
Biotin-detection antibody (Concentrated) 60ul/120ul 4°C
Antibody dilution buffer 5ml/10ml 4°C
HRP-Streptavidin Conjugate(SABC) 60ul/120ul 4°C (light sensitive)
SABC dilution buffer 5ml/10ml 4°C TMB substrate 5ml/10ml 4°C (light sensitive)
Stop solution 5ml/10ml 4°C
Wash buffer (25X) 15ml/30ml 4°C

Protocol Summary

1. Wash plate 2 times before adding standard, sample and control (zero) wells!

2. Add 100μL standard or sample to each well and incubate for 90 minutes at 37°C

3. Aspirate and wash plates 2 times

4. Add 100μL Biotin-labeled antibody working solution to each well and incubate for 60 minutes at 37°C

5. Add 100 μLSABC Working Solution into each well and incubate for 30 minutes at 37°C

6. Aspirate and wash plates 5 times

7. Add 90μL TMB Substrate. Incubate 15 -30 minutes at 37°C

8. Add 50μL Stop Solution. Read at 450nm immediately

9. Calculation

Shipping: Overnight, 2-day with ice pack, international shipping

Storage: 4^oC

Safety: Contains sulfuric acid. TMZ. Avoid contact. Wear eye protection, gloves

Category	Download Link
Protocol Manual	click here
MSDS	click here

Human Soluble Alpha Klotho (SAKL, α-Klotho) ELISA Kit (Part hklotho-ELISA)
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Human Soluble α-Klotho ELISA Kit (alpha-klotho, alpha klotho, 96T; 27998)
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$773.00

Tel: 1.858.395.4026; or 1.800.462.4507
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