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Scientific Background
This ELISA kit applies to the in vitro quantitative determination of Human KL concentrations in serum, plasma and other biological fluids.
Assay Principle
This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human KL. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human KL and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human KL, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human KL. You can calculate the concentration of Human KL in the samples by comparing the OD of the samples to the standard curve.
Performance Specifications
| Sensitivity | 0.19 ng/mL |
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| Detection Range | 0.31-20 ng/mL |
| Total Assay Time | 3 h 30 min |
| Compatible Sample Types | Serum, plasma and other biological fluids |
| Species Reactivity | Human |
| Detection Method | Sandwich |
| Precision (CV) | Both intra-CV and inter-CV are < 10%. |
| Recovery Rate | 80%-120% |
| Storage | 2-8℃,12 months |
✓ Research-Grade Validation
Specificity
This kit recognizes Human KL in samples. No significant cross-reactivity or interference between Human KL and analogues was observed
Safety & Regulatory
Handle reagents in accordance with institutional biosafety guidelines. Refer to the Safety Data Sheet (SDS) for complete hazard and handling information. Contains components that may require special disposal procedures per local regulations.
This kit is validated for use with Serum, plasma and other biological fluids. For unlisted matrices (e.g., tissue lysate, urine), perform a spike-and-recovery experiment to confirm assay performance before generating reportable data. Sample dilution in the kit's provided diluent is recommended to minimize matrix interference.
The minimum detectable concentration (sensitivity) of this kit is 0.19 ng/mL. Values below this threshold should be reported as below the limit of detection (<LOD) and should not be extrapolated from the standard curve.
The total assay time from sample addition to absorbance reading is approximately 3 h 30 min, including all incubation, wash, and substrate steps. Hands-on time is typically 1–2 hours; most steps involve passive plate incubation. Plan the assay as a single uninterrupted session for best results.
Standard components of this Sandwich ELISA Kit typically include: pre-coated microplate (96-well strip format), lyophilized or liquid recombinant KL standard, detection antibody, streptavidin-HRP conjugate, TMB substrate, stop solution, wash buffer concentrate, and sample/standard diluent. Refer to the kit insert or datasheet for the exact component list and storage requirements.
This kit uses colorimetric (TMB/HRP) detection and requires a standard microplate absorbance reader capable of measuring at 450 nm. A reference wavelength of 570 nm or 630 nm is recommended to reduce background. No specialized fluorescence or luminescence reader is needed. Ensure the instrument is calibrated and the plate is clean and free of condensation before reading.
Tissue samples may contain endogenous or exogenous proteases during processing, leading to degradation of extracted proteins. Therefore, it's necessary to add protease inhibitors during processing to ensure the integrity of target proteins. If customers can keep samples cold and handle them quickly during processing, omitting the protease inhibitor may not have a significant effect. After preparation, samples should be tested promptly or immediately aliquoted and frozen at -20°C or -80°C.
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AB0240?CORRELATION BETWEEN KLOTHO LEVELS AND ANKLE-BRACHIAL INDEX IN FEMALE RHEUMATOID ARTHRITIS PATIENTS (2022) ANNALS OF THE RHEUMATIC DISEASES. 10.1136/annrheumdis-2022-eular.446
Efficacy of a novel proprietary dietary supplement (TRI 360TM) on psychological symptoms and stress-related quality of life in adult subjects: A randomized controlled clinical trial (2022) Frontiers in Psychiatry. 10.3389/fpsyt.2022.919284
Renal lipid accumulation and aging linked to tubular cells injury via ANGPTL4 (2024) MECHANISMS OF AGEING AND DEVELOPMENT. 10.1016/j.mad.2024.111932
CCL5 Suppresses Klotho Expression via p-STAT3/DNA Methyltransferase1-Mediated Promoter Hypermethylation (2022) Frontiers in Physiology. 10.3389/fphys.2022.856088
Kernelized k-Local Hyperplane Distance Nearest-Neighbor Model for Predicting Cerebrovascular Disease in Patients With End-Stage Renal Disease (2021) Frontiers in Neuroscience. 10.3389/fnins.2021.773208
Body composition, adipokines, FGF23-Klotho and bone in kidney transplantation: Is there a link? (2021) JOURNAL OF NEPHROLOGY. 10.1007/s40620-021-00972-9
Predictive value of bone metabolism markers in the progression of diabetic kidney disease: a cross-sectional study (2024) Frontiers in Endocrinology. 10.3389/fendo.2024.1489676
The FGF23–Klotho axis promotes microinflammation in chronic kidney disease (2024) CYTOKINE. 10.1016/j.cyto.2024.156781
Alpha-Klotho protein in systemic lupus erythematosus (2023) CLINICAL AND EXPERIMENTAL RHEUMATOLOGY. 10.55563/clinexprheumatol/salqon
Relationship between klotho, neurotrophic factors (BDNF, NGF, GDNF) and cognitive functions in patients with bipolar disorder (2025) BMC Psychiatry. 10.1186/s12888-025-06469-0
The roles of Klotho and FGF-23 in bipolar manic episode (2023) European Review for Medical and Pharmacological Sciences. 10.26355/eurrev_202310_34078
A Comprehensive Study on the Anti-cancer Effects of Quercetin and Its Epigenetic Modifications in Arresting Progression of Colon Cancer Cell Proliferation (2023) ARCHIVUM IMMUNOLOGIAE ET THERAPIAE EXPERIMENTALIS. 10.1007/s00005-023-00669-w
An evaluation of both serum Klotho/FGF-23 and apelin-13 for detection of diabetic nephropathy (2023) Hormones-International Journal of Endocrinology and Metabolism. 10.1007/s42000-023-00464-4
Enhancement of Apo2L/TRAIL signaling pathway receptors by the?activation of Klotho gene?with?CRISPR/Cas9 in Caco-2 colon cancer cells (2021) MEDICAL ONCOLOGY. 10.1007/s12032-021-01595-7
Development of a nomogram for predicting incident heart failure and all-cause mortality in patients with chronic kidney disease: a 3-year follow-up study (2026) Frontiers in Medicine. 10.3389/fmed.2026.1784717
The Association of Fibroblast Growth Factor 23 with Arterial Stiffness and Atherosclerosis in Patients with Autosomal Dominant Polycystic Kidney Disease (2018) KIDNEY & BLOOD PRESSURE RESEARCH. 10.1159/000492244
The correlation between fibroblast growth factor-23 and ESRD patients with hearing impairment (2021) PeerJ. 10.7717/peerj.12295
Comparative Diagnostic Performance of Serum α-Klotho and FGF-23 in Predicting Obstructive Sleep Apnea Severity: A Novel Biomarker Approach (2026) Journal of Clinical Medicine. 10.3390/jcm15062316
Association of plasma neutrophil gelatinase-associated lipocalin with parameters of CKD–MBD in maintenance hemodialysis patients (2021) JOURNAL OF BONE AND MINERAL METABOLISM. 10.1007/s00774-021-01248-9
Association of plasma neutrophil gelatinase-associated lipocalin and thoracic aorta calcification in maintenance hemodialysis patients with and without diabetes (2022) BMC Nephrology. 10.1186/s12882-022-02773-z
Baseline fibroblast growth factor 23 predicts incident heart failure and cardiovascular mortality in patients with chronic kidney disease: A 3-year follow-up study (2024) IJC Heart & Vasculature. 10.1016/j.ijcha.2024.101587
A Self-Representation-Based Fuzzy SVM Model for Predicting Vascular Calcification of Hemodialysis Patients (2021) Computational and Mathematical Methods in Medicine. 10.1155/2021/2464821
The Relationship Between Serum Fibroblast Growth Factor 23 and Klotho Protein and Low Bone Mineral Density in Middle-Aged and Elderly Patients with End-Stage Renal Disease (2023) HORMONE AND METABOLIC RESEARCH. 10.1055/a-2168-5089
Effects of acute aerobic exercise on cytokines, klotho, irisin, and vascular endothelial growth factor responses in rheumatoid arthritis patients (2022) IRISH JOURNAL OF MEDICAL SCIENCE. 10.1007/s11845-022-02970-7
Klotho, vitamin D and homocysteine levels during acute episode and remission periods in schizophrenia patients (2019) NORDIC JOURNAL OF PSYCHIATRY. 10.1080/08039488.2019.1582697
