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Human Soluble Alpha Klotho (SAKL, α-Klotho) ELISA Kit (Part hklotho-ELISA)

Human Soluble Alpha Klotho (SAKL, α-Klotho) ELISA Kit (Part hklotho-ELISA)
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Price: $685.00
Availability: In Stock
Model: hklotho-elisa

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Human Soluble alpha-Klotho (SAKL) ELISA Kit (96T) 

   Protocol

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.  


Product name

Human SAKL(Soluble alpha-Klotho) ELISA Kit
Catalogue No.

hklotho-ELISA
Detection method

Sandwich ELISA, Double Antibody
Size

96T
Range

93.75-6000pg/ml
Sensitivity

< 6.15pg/ml
Standard curve

Human SAKL(Soluble alpha-Klotho) ELISA Kit
Storage

4 ℃ for 6 months
Recovery

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

Matrix Recovery range(%) Average(%)
serum(n=5) 85-101 92
EDTA plasma(n=5) 91-100 98
heparin plasma(n=5) 86-103 94
Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of 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) 87-104% 89-105% 85-105% 92-104%
EDTA plasma(n=5) 83-91% 86-100% 82-99% 87-97%
heparin plasma(n=5) 86-98% 80-97% 80-98% 80-94%
CV(%)

Intra-Assay: CV<8%
Inter-Assay: CV<10%
 
 
Kit Components Item Specifications (48T/96T) 
  1. Storage Micro ELISA Plate (Dismountable) 8×6 or 8×12 4°C/-20°C 
  2. Lyophilized Standard 1 vial or 2 vial 4°C/-20°C Sample 
  3. Standard dilution buffer 10ml/20ml 4°C 
  4. Biotin-detection antibody (Concentrated) 60ul/120ul 4°C 
  5. Antibody dilution buffer 5ml/10ml 4°C 
  6. HRP-Streptavidin Conjugate(SABC) 60ul/120ul 4°C (light sensitive) 
  7. SABC dilution buffer 5ml/10ml 4°C TMB substrate 5ml/10ml 4°C (light sensitive) 
  8. Stop solution 5ml/10ml 4°C 
  9. 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:  4oC
Safety:  Contains sulfuric acid.  TMZ.  Avoid contact.  Wear eye protection, gloves


 

Category Download Link
Protocol Manual click here
MSDS click here