Human Klotho ELISA Kit

Code CSB-E13235h
Size 96T,5×96T,10×96T
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Product Details

Target Name
klotho
Alternative Names
Kl ELISA Kit; KLOT_HUMAN ELISA Kit; Klotho peptide ELISA Kit
Abbreviation
KL
Uniprot No.
Species
Homo sapiens (Human)
Sample Types
serum, plasma, tissue homogenates
Detection Range
0.156 ng/mL-10 ng/mL
Sensitivity
0.039 ng/mL
Assay Time
1-5h
Sample Volume
50-100ul
Detection Wavelength
450 nm
Research Area
Developmental Biology
Assay Principle
quantitative
Measurement
Sandwich
Precision
Intra-assay Precision (Precision within an assay): CV%<8%      
Three samples of known concentration were tested twenty times on one plate to assess.  
Inter-assay Precision (Precision between assays): CV%<10%      
Three samples of known concentration were tested in twenty assays to assess.    
             
Linearity
To assess the linearity of the assay, samples were spiked with high concentrations of human Klotho in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay.
  Sample Serum(n=4)  
1:1 Average % 89  
Range % 86-92  
1:2 Average % 98  
Range % 91-105  
1:4 Average % 96  
Range % 92-102  
1:8 Average % 93  
Range % 86-99  
Recovery
The recovery of human Klotho spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay as directed in the Sample Preparation section.
Sample Type Average % Recovery Range  
Serum (n=5) 94 89-98  
EDTA plasma (n=4) 97 90-102  
             
             
Typical Data
These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.
ng/ml OD1 OD2 Average Corrected  
10 2.491 2.452 2.472 2.350  
5 2.104 2.076 2.090 1.968  
2.5 1.504 1.562 1.533 1.411  
1.25 0.869 0.931 0.900 0.778  
0.625 0.494 0.507 0.501 0.379  
0.312 0.292 0.279 0.286 0.164  
0.156 0.185 0.179 0.182 0.060  
0 0.126 0.117 0.122    
Materials provided
  • A micro ELISA plate ---The 96-well plate has been pre-coated with an anti-human klotho antibody. This dismountable microplate can be divided into 12 x 8 strip plates.
  • Two vials lyophilized standard ---Dilute a bottle of the standard at dilution series, read the OD values, and then draw a standard curve.
  • One vial Biotin-labeled klotho antibody (100 x concentrate) (120 μl/bottle) ---Act as the detection antibody.
  • One vial HRP-avidin (100 x concentrate) (120 μl/bottle) ---Bind to the detection antibody and react with the TMB substrate to make the solution chromogenic.
  • One vial Biotin-antibody Diluent (15 ml/bottle) ---Dilute the Biotin-antibody.
  • One vial HRP-avidin Diluent (15 ml/bottle) ---Dilute the HRP-avidin solution.
  • One vial Sample Diluent (50 ml/bottle)---Dilute the sample to an appropriate concentration.
  • One vial Wash Buffer (25 x concentrate) (20 ml/bottle) ---Wash away unbound or free substances.
  • One vial TMB Substrate (10 ml/bottle) ---Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
  • One vial Stop Solution (10 ml/bottle) ---Stop the color reaction. The solution color immediately turns from blue to yellow.
  • Four Adhesive Strips (For 96 wells) --- Cover the microplate when incubation.
  • An instruction manual
Materials not provided
  • A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm or 570 nm.
  • An incubator can provide stable incubation conditions up to 37°C±5°C.
  • Centrifuge
  • Vortex
  • Squirt bottle, manifold dispenser, or automated microplate washer
  • Absorbent paper for blotting the microtiter plate
  • 50-300ul multi-channel micropipette
  • Pipette tips
  • Single-channel micropipette with different ranges
  • 100ml and 500ml graduated cylinders
  • Deionized or distilled water
  • Timer
  • Test tubes for dilution
Troubleshooting
and FAQs
Storage
Store at 2-8°C. Please refer to protocol.
Lead Time
3-5 working days after you place the order, and it takes another 3-5 days for delivery via DHL or FedEx
Description

This Human Klotho ELISA Kit was designed for the quantitative measurement of Human Klotho protein in serum, plasma, tissue homogenates. It is a Sandwich ELISA kit, its detection range is 0.156 ng/mL-10 ng/mL and the sensitivity is 0.039 ng/mL .

Citations

Customer Reviews and Q&A

 Customer Reviews

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 Q&A
Q:

Does the kit come with the QCs included? Would it be possible to have a control sample for this kit? Sorry for the confusion, that’s what they meant for QC

A:
Thanks for your inquiry.
The standard of this kit plays the role as quality control.
So sorry that there is no separate quality control sample to provide.

Target Background

Function
(From Uniprot)
May have weak glycosidase activity towards glucuronylated steroids. However, it lacks essential active site Glu residues at positions 239 and 872, suggesting it may be inactive as a glycosidase in vivo. May be involved in the regulation of calcium and phosphorus homeostasis by inhibiting the synthesis of active vitamin D. Essential factor for the specific interaction between FGF23 and FGFR1.; The Klotho peptide generated by cleavage of the membrane-bound isoform may be an anti-aging circulating hormone which would extend life span by inhibiting insulin/IGF1 signaling.
Gene References into Functions
  1. sKlotho is a biomarker of renal dysfunction and a potential treatment target for renoprotection in T2 diabetes mellitus. PMID: 29509906
  2. Soluble klotho levels were inversely related to brain atrophy in alcoholic patients. PMID: 29846497
  3. The tissue klotho protein expression was associated with a lower risk and progression of malignancies. Klotho may be a protective factor against malignancies risk/progression. PMID: 29775618
  4. Serum Klotho may be a novel risk factor for atherosclerosis in maintenance hemodialysis patients PMID: 30064142
  5. a single maximal aerobic exercise session of 20 min duration induced serum levels of klotho, particularly in women PMID: 27251899
  6. Knockdown of klotho gene expression normalized IGF-1R and Wnt1 protein expressions and Akt phosphorylation in granulosa cells from patients with PCOS; it also blocked the effects of insulin on apoptosis and proliferation in granulosa cells. PMID: 28673204
  7. Schizophrenia patients, KL-VS has a selective effect on memory, with heterozygotes in CD and CS clusters performing worse than non-carriers PMID: 28673754
  8. The change in concentration of soluble alpha-klotho during the 1.5-years follow-up was an indicator of CKD progression. Renal damage associated with a reduction of alpha-klotho may involve the upregulation of plasma aldosterone. PMID: 29843135
  9. Cord blood klotho levels were inversely correlated with leptin and insulin levels at birth PMID: 29648999
  10. solubility-Klotho level was closely correlated with kidney function PMID: 29061648
  11. To evaluate the role of klotho in the pathogenesis of systemic sclerosis, its serum concentration was measured in patients compared to healthy controls, and its association with cutaneous and visceral involvement was assessed. Spearman's test showed no significant association between klotho serum levels and systemic sclerosis activity, concerning either clinical, laboratory or instrumental findings. PMID: 29320841
  12. Klotho may play an inhibiting role in lipopolysaccharide-induced inflammation injury by inhibiting NF-kappaB and Wnt signaling pathways in HK-2 cells. PMID: 29441993
  13. High klotho expression is associated with renal fibrosis and podocyte injuries. PMID: 29590173
  14. Although our study revealed higher serum ET-1 and lower serum alpha-Klotho levels in systemic sclerosis patients compared to healthy controls, there were not any significant correlations between their serum levels with severity of organ involvement. PMID: 28831601
  15. The A allele of the G395A polymorphism of Klotho, which emerges the higher levels of phosphate, may be associated with the risk of mortality in Iranian patients receiving hemodialysis. PMID: 29190606
  16. This study showed that A reduced blood alpha-Klotho level was correlated with grading of cerebral Deep White Matter Lesions and was accompanied by cognitive decline as an independent risk factor. PMID: 29154273
  17. Klotho deficiency is not just a biomarker but a contributing factor to the development of acute kidney injury. PMID: 29550818
  18. Forced expression of Klotho resulted in decline of activation of IGF-1R signaling, accompanied by decreased phosphorylation of its downstream targets, including AKT and ERK1/2. These data indicated that Klotho acts as a tumor suppressor via inhibiting IGF-1R signaling, thus suppressing the viability and promoting apoptosis in Tcell lymphoma. PMID: 28656297
  19. serum levels increase following kidney transplantation, although changes are modest compared to more significant improvements in renal function and iFGF-23 levels PMID: 28120476
  20. The functional KL-VS variant of the KLOTHO gene protects against the development of retinopathy in patients with T1 diabetes. PMID: 29247834
  21. the immunohistochemical expressions of Klotho and DNMT3a in tissues obtained from oral dysplasia and oral squamous cell carcinoma, is reported. PMID: 28303350
  22. Klotho suppresses Sox9 upregulation and intranuclear translocation. Klotho inhibits high phosphate-induced osteogenic activity in human aortic valve interstitial cells. PMID: 28332126
  23. Data suggest that serum levels of alpha-klotho and beta-klotho are down-regulated in patients with type 2 diabetes, regardless of diabetic complications. PMID: 27916483
  24. Low expression levels of Klotho were observed in DLBCL patients and cell lines. Enforced expression of Klotho could significantly induce cell apoptosis and inhibit tumor growth in DLBCL. Upregulation of Klotho resulted in declined activation of IGF-1R signaling, accompanied with decreased phosphorylation of its downstream targets, including AKT and ERK1/2. PMID: 28153033
  25. Data suggest that plasma alpha-klotho is down-regulated in subjects with type 2 diabetes as diabetic nephropathy progresses; plasma alpha-klotho level (not urinary alpha-klotho level) may be an early biomarker for predicting renal impairment in type 2 diabetes. PMID: 27037042
  26. Hmgb1 can inhibit Klotho gene expression and malignant phenotype in melanoma cells through activation of NF-kappaB signaling. PMID: 27779100
  27. patients with autosomal dominant polycystic kidney disease may potentially have different soluble alpha-Klotho production/clearance characteristics than the patients with other parenchymal renal diseases. PMID: 27733443
  28. KL and SFRP1 methylation were more predominant in nasopharyngeal tumors. PMID: 28812223
  29. The beta-Klotho gene expresses only in retina and optic nerve. The gamma-Klotho gene may regulate human lens epithelial cells' function mainly by the MAPK/ERK1/2 signaling pathway. PMID: 28095050
  30. Down-Regulation of Soluble alpha-Klotho is Associated with Reduction in Serum Irisin Levels in Chronic Obstructive Pulmonary Disease. soluble a-klotho is one possible factor involved in reduced irisin release from skeletal muscle PMID: 27140192
  31. Klotho is a co-receptor for FGF23. Together they play an important role in bone mineral and vitamin D metabolism. In chronic kidney disease, disturbances in bone metabolism increase cardiovascular risk. In the kidney, klotho induces phosphaturia. Klotho levels drop in chronic kidney disease. It inhibits vascular calcification. Review. PMID: 27118192
  32. A single copy of klotho allele KL-VS is associated with longevity, better health, increased cognition, and bigger regional brain volume. PMID: 28431289
  33. Review of Klotho function and dysfunction in mice and humans. [Review Article] PMID: 27576165
  34. The Klotho KL-VS variant is associated with an increase in the incidence of dementia in older men, in a dose-dependent fashion (intermediate for heterozygosis and highest for homozygosis). PMID: 28539162
  35. Our findings provide the first evidence that alpha-Klotho is significantly decreased in the microvasculature in systemic sclerosis skin PMID: 28183357
  36. indicates an inverse interrelationship between inflammation and Klotho in atherosclerosis PMID: 28963437
  37. Administration of soluble klotho significantly reduced obstruction-induced renal fibrosis in wild-type mice, but not in Trpc6 knockout mice, indicating that klotho and TRPC6 inhibition act in the same pathway to protect against obstruction-induced renal fibrosis. PMID: 27979597
  38. The role of Klotho protein in aging and central nervous system pathology is reviewed. PMID: 28988528
  39. These results support the notion that sialogangliosides and lipid rafts are membrane receptors for sKlotho and that the KL1 domain is sufficient for the tested biologic activities. PMID: 28442546
  40. Klotho is expressed in human dermal fibroblasts and dermal microvascular endothelial cells and is implicated in the process of angiogenesis. PMID: 27260080
  41. A factor for functioning of the brain in aged is KLOTHO gene. PMID: 26405063
  42. Data suggest Klotho attenuates ox-LDL- (oxidized low density lipoprotein)-induced oxidative stress in vascular endothelial cells via up-regulation of oxidative scavengers (lipoprotein and nitric oxide), activation of the PI3K/Akt/eNOS signaling, and down-regulation of LOX1 (lectin-like ox-LDL receptor) expression. (PI3K = phosphatidylinositol 3-Kinase; Akt = proto-oncogene c-Akt; eNOS = endothelial nitric oxide synthase) PMID: 28407763
  43. Targeting klotho may thus represent a therapeutic strategy . PMID: 27696667
  44. FGF23 affected NO synthesis and oxidative stress in human coronary artery endothelial cells are modulated by Klotho. PMID: 27448998
  45. Conversely, siRNA-mediated Klotho silencing up-regulated Egr-1, FN, and Col I expression and the p-Smad3/Smad3 ratio. Moreover, the effects of si-Klotho on Egr-1 expression were abolished by the TGF-beta1 inhibitor SB-431542. Klotho overexpression can prevent mesangial ECM production in high-glucose-treated human MCs, an effect that has been partially attributed to Egr-1 down-regulation facilitated by TGF-beta1/Smad3 s... PMID: 28411025
  46. Klotho levels were higher in acute kidney injury patients and females and lower in end stage renal disease patients than in healthy adults and patients with moderate chronic kidney disease. PMID: 28157067
  47. Genetic polymorphisms such as the G-395A polymorphism in the promoter region of the Klotho gene have been associated with the development of essential hypertension. PMID: 27215557
  48. Klotho was demonstrated to act as a potent tumor suppressor in human ovarian cancer cells. Reduced Klotho expression was detected in the specimens of patients with ovarian cancer, and overexpression of Klotho significantly inhibited cell proliferation of human ovarian cancer cells. PMID: 28259911
  49. sKlotho is increased in active acromegaly and normalises after successful treatment. PMID: 27387242
  50. Any polymorphism altering the function of klotho gene may result with stone formation. We found that there are more GG sequences of G395A gene in patients with urinary tract stone disease. That may be a polymorphism of klotho gene which results with stone formation. PMID: 26597587

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Involvement in disease
Tumoral calcinosis, hyperphosphatemic, familial (HFTC)
Subcellular Location
[Isoform 1]: Cell membrane; Single-pass type I membrane protein. Apical cell membrane; Single-pass type I membrane protein.; [Isoform 2]: Secreted.; [Klotho peptide]: Secreted.
Protein Families
Glycosyl hydrolase 1 family, Klotho subfamily
Tissue Specificity
Present in cortical renal tubules (at protein level). Soluble peptide is present in serum and cerebrospinal fluid. Expressed in kidney, placenta, small intestine and prostate. Down-regulated in renal cell carcinomas, hepatocellular carcinomas, and in chro
Database Links

HGNC: 6344

OMIM: 211900

KEGG: hsa:9365

STRING: 9606.ENSP00000369442

UniGene: Hs.524953

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