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Multiplex Assay Kit for Klotho (KL) ,etc. by FLIA (Flow Luminescence Immunoassay)
KLa; Klotho Alpha
(Note: Up to 8-plex in one testing reaction)
- Product No.LMH757Ra
- Organism SpeciesRattus norvegicus (Rat) Same name, Different species.
- Sample TypeSerum, plasma, tissue homogenates and other biological fluids
- Test MethodDouble-antibody Sandwich
- Assay Length3.5h
- Detection Range1.95-2000pg/mL
- SensitivityThe minimum detectable dose of this kit is typically less than 0.65 pg/mL.
- DownloadInstruction Manual
- UOM 8Plex 7Plex 6Plex 5Plex 4Plex 3Plex 2Plex1Plex
- FOB
US$ 474
US$ 492
US$ 520
US$ 556
US$ 593
US$ 648
US$ 730
Result
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Specificity
This assay has high sensitivity and excellent specificity for detection of Klotho (KL) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Klotho (KL) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.
Recovery
Matrices listed below were spiked with certain level of recombinant Klotho (KL) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Klotho (KL) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.
Matrix | Recovery range (%) | Average(%) |
serum(n=5) | 86-101 | 92 |
EDTA plasma(n=5) | 78-96 | 80 |
heparin plasma(n=5) | 84-98 | 89 |
sodium citrate plasma(n=5) | 80-93 | 89 |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Klotho (KL) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Klotho (KL) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%
Linearity
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Klotho (KL) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) | 95-102% | 78-91% | 89-97% | 79-90% |
EDTA plasma(n=5) | 80-96% | 86-101% | 89-104% | 80-93% |
heparin plasma(n=5) | 92-105% | 87-95% | 97-105% | 80-89% |
sodium citrate plasma(n=5) | 78-102% | 97-105% | 80-90% | 79-94% |
Stability
The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.
Reagents and materials provided
Reagents | Quantity | Reagents | Quantity |
96-well plate | 1 | Plate sealer for 96 wells | 4 |
Pre-Mixed Standard | 2 | Standard Diluent | 1×20mL |
Pre-Mixed Magnetic beads (22#:KL) | 1 | Analysis buffer | 1×20mL |
Pre-Mixed Detection Reagent A | 1×120μL | Assay Diluent A | 1×12mL |
Detection Reagent B (PE-SA) | 1×120μL | Assay Diluent B | 1×12mL |
Sheath Fluid | 1×10mL | Wash Buffer (30 × concentrate) | 1×20mL |
Instruction manual | 1 |
Assay procedure summary
1. Preparation of standards, reagents and samples before the experiment;
2. Add 100μL standard or sample to each well,
add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.
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Magazine | Citations |
Life Sciences | High phosphate feeding induced arterial medial calcification in uremic rats: Roles of Lanthanum carbonate on protecting vasculature Pubmed: 24012609 |
EXPERIMENTAL & CLINICAL CARDIOLOGY | Sick Sinus Syndrome May Attribute to the Decrease of ?±-klotho Protein in Mice Com:Source |
Int J Clin Exp Pathol.? | α-Klotho is an acute phase protein and altered by restraint stress in mice Pubmed:Pmc4203207 |
Annales de Biologie Clinique | Les biomarqueurs des calcifications vasculaires PubMed: 26069069 |
Mol Med Rep | Effect of secondary hyperparathyroidism serum on endothelial cells and intervention with Klotho PubMed: 25873020 |
Calcif Tissue Int | Human Preosteoblastic Cell Culture from a Patient with Severe Tumoral Calcinosis-Hyperphosphatemia Due to a NewGALNT3 PubMed: 25899975 |
中华内分泌代谢杂志 | 抗衰老蛋白Klotho 阻断高糖培养的大鼠肾小球系膜细胞TLR4/NF-κB p65/NGAL 通路的激活及其作用机制 Cn: Article |
Mol Cell Endocrinol | The function of miR-199a-5p/Klotho regulating TLR4/NF-κB p65/NGAL pathways in rat mesangial cells cultured with high glucose and the mechanism PubMed: 26419931 |
Arch Med Sci | Soluble Klotho protein as a novel serum biomarker in patients with acromegaly pmc:PMC4754356 |
Am J Nephrol | Klotho Deficiency Aggravates Tacrolimus-Induced Renal Injury via the Phosphatidylinositol 3-Kinase-Akt-Forkhead Box Protein O Pathway Pubmed:27174564 |
Sci Rep. | Klotho preservation via histone deacetylase inhibition attenuates chronic kidney disease-associated bone injury in mice. pubmed:28387374 |
Nephrology | Is there a role for newer biomarkers in chronic kidney disease-mineral and bone disorder management? pubmed:28429560 |
Journal of Integrative Nephrology & Andrology | Associations between serum fibroblast growth factor 23 level and intradialytic hypotension in hemodialysis patients issn:2394-2916 |
International Journal of Clinical and Experimental Pathology | Protective effect of restructuring klotho protein combined with trimetazidine on myocardial injury in mice ISSN:1936-2625 |
Journal of veterinary medical science | Changes of Klotho protein and Klotho mRNA expression in a hydroxy-L-proline induced hyperoxaluric rat model pubmed:28943597 |
Cellular Physiology and Biochemistry | Klotho Reduces Necroptosis by Targeting Oxidative Stress Involved in Renal Ischemic-Reperfusion Injury Pubmed:29550818 |
American Journal of Translational Research | Advanced maternal age impairs spatial learning capacity in young adult mouse offspring Pubmed:29636887 |
Journal of Diabetes and its Complications | Clinical implication of alterations in serum Klotho levels in patients with type 2 diabetes mellitus and its associated complications Pubmed:30042059 |
Medical Science Monitor | Klotho Inhibits Proliferation and Migration of Angiotensin II-Induced Vascular Smooth Muscle Cells (VSMCs) by Modulating NF-κB p65, Akt, and Extracellular Signal … Pubmed:30004089 |
Nephrology Dialysis Transplantation | High-serum phosphate and parathyroid hormone distinctly regulate bone loss and vascular calcification in experimental chronic kidney disease Pubmed: 30189026 |
BMC Nephrology | The role of klotho in chronic kidney disease Pubmed: 30348110 |
Nature Communications | Age-related declines in α-Klotho drive progenitor cell mitochondrial dysfunction and impaired muscle regeneration Pubmed: 30451844 |
Kidney International | Wnt/β-catenin signaling mediates both heart and kidney injury in type 2 cardiorenal syndrome Pubmed: 30770217 |
Expert Opinion on Therapeutic Targets | The use of LP533401 as a therapeutic option for renal osteodystrophy affects, renal calcium handling, vitamin D metabolism and bone health in uremic rats Pubmed: 30801205 |
Mol Immunol | CD317 mediates immunocytolysis resistance by RICH2/cytoskeleton-dependent membrane protection Pubmed: 33223223 |
Journal of Clinical Hypertension | Effects of salt intervention on serum levels of Klotho influenced by salt sensitivity Pubmed: 33164306 |
CURRENT EYE RESEARCH | Expression of α-Klotho is downregulated and associated with oxidative stress in the lens in streptozotocin-induced diabetic rats Pubmed: 32744464 |
international immunopharmacology | Klotho ameliorates the onset and progression of cataract via suppressing oxidative stress and inflammation in the lens in streptozotocin-induced diabetic rats Pubmed: 32438076 |
Purinergic Signalling | P2X7 siRNA targeted to the kidneys increases klotho and delays the progression of experimental diabetic nephropathy. Pubmed: 32377919 |
Oxidative Medicine and Cellular Longevity | Ameliorating Effect of Klotho Protein on Rat Heart during I/R Injury Pubmed: 33123314 |
NEUROSCIENCE | Increased Plasma Level of Longevity Protein Klotho as a Potential Indicator of Cognitive Function Preservation in Patients With Schizophrenia Pubmed: 32612508 |
Pharmacological Reports | ¦ÁKlotho protein has therapeutic activity in contrast-induced acute kidney injury by limiting NLRP3 inflammasome-mediated pyroptosis and promoting autophagy 33675964 |
J Mol Neurosci | Investigation of the Role of Induced Overexpression of the Isolated Secreted Klotho on the A-172 Human Glioblastoma Cells Pubmed:35112226 |
Reproductive Biology and Endocrinology | Expression of Klotho in Follicles of Patients With Polycystic Ovary Syndrome and Its Clinical Significance |
Journal of Ovarian Research | Klotho is a prognostic indicator of oocytes qualities in polycystic ovary syndrome |
Catalog No. | Related products for research use of Rattus norvegicus (Rat) Organism species | Applications (RESEARCH USE ONLY!) |
APH757Ra01 | Active Klotho (KL) | Cell culture; Activity Assays. |
RPH757Ra02 | Recombinant Klotho (KL) | Positive Control; Immunogen; SDS-PAGE; WB. |
RPH757Ra01 | Recombinant Klotho (KL) | Positive Control; Immunogen; SDS-PAGE; WB. |
PAH757Ra02 | Polyclonal Antibody to Klotho (KL) | WB; IHC; ICC; IP. |
PAH757Ra01 | Polyclonal Antibody to Klotho (KL) | WB; IHC; ICC; IP. |
MAH757Ra21 | Monoclonal Antibody to Klotho (KL) | WB; IHC; ICC; IP. |
SEH757Ra | ELISA Kit for Klotho (KL) | Enzyme-linked immunosorbent assay for Antigen Detection. |
LMH757Ra | Multiplex Assay Kit for Klotho (KL) ,etc. by FLIA (Flow Luminescence Immunoassay) | FLIA Kit for Antigen Detection. |