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Human sRANKL(Soluble Receptor Activator of Nuclear factor-kB Ligand) ELISA Kit

Citations(12)Uniprot : O14788

Cat.No.:E-EL-H5558
Reactivity: Human

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Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T/48T
Assay time	3.5h
Detection range	15.63-1000 pg/mL
Sensitivity	9.38 pg/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 100μL
Specificity	This kit recognizes Human sRANKL in samples. No significant cross-reactivity or interference between Human sRANKL and analogues was observed.
Reproducibility	Both intra-CV and inter-CV are < 10%.
Application	This ELISA kit applies to the in vitro quantitative determination of Human sRANKL concentrations in serum, plasma and other biological fluids.

Test 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 sRANKL. 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 sRANKL 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 sRANKL, 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 sRANKL. You can calculate the concentration of Human sRANKL in the samples by comparing the OD of the samples to the standard curve.

Kit components & Storage

An unopened kit can be stored at 2-8℃ for 1 month. If the kit is not supposed to be used within 1 month, store the components separately according to the following conditions once the kit is received.

Item	Specifications	Storage
Micro ELISA Plate(Dismountable)	96T: 8 wells ×12 strips 48T: 8 wells ×6 strips	-20℃, 6 months
Reference Standard	96T: 2 vials 48T: 1 vial
Concentrated Biotinylated Detection Ab (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL
Concentrated HRP Conjugate (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL	-20℃(Protect from light), 6 months
Reference Standard & Sample Diluent	1 vial, 20 mL	2-8°C, 6 months
Biotinylated Detection Ab Diluent	1 vial, 14 mL
HRP Conjugate Diluent	1 vial, 14 mL
Concentrated Wash Buffer (25×)	1 vial, 30 mL
Substrate Reagent	1 vial, 10 mL	2-8℃(Protect from light)
Stop Solution	1 vial, 10 mL	2-8°C
Plate Sealer	5 pieces
Manual	1 copy
Certificate of Analysis	1 copy

Technical Data

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human sRANKL were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human sRANKL were tested on 3 different plates, 20 replicates in each plate.

	Intra-assay Precision			Inter-assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	20	20	20
Mean (pg/mL)	52.28	122.42	372.33	55.30	121.01	370.30
Standard deviation	3.20	5.66	17.61	3.65	5.14	16.11
CV (%)	6.12	4.62	4.73	6.60	4.25	4.35

Recovery

The recovery of Human sRANKL spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample Type	Range (%)	Average Recovery (%)
Serum(n=8)	92-104	99
EDTA plasma (n=8)	88-101	93
Cell culture media (n=8)	92-102	97

Linearity

Samples were spiked with high concentrations of Human sRANKL and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

Database Links	SwissProt: O14788
Research Area	Cancer, Cardiovascular, Immunology

Assay Procedures

	1. Add 100μL standard or sample to the wells. Incubate for 90 min at 37°C
	2. Discard the liquid, immediately add 100μL Biotinylated Detection Ab working solution to each well. Incubate for 60 min at 37°C
	3. Aspirate and wash the plate for 3 times
	4. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
	5. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
	6. Add 50μL Stop Solution
	7. Read the plate at 450nm immediately. Calculation of the results

Citations

Frontiers in Oncology (2021) IF: 6.244
Single-Cell Transcriptomics Reveals the Complexity of the Tumor Microenvironment of Treatment-Naive Osteosarcoma

DOI: 10.3389/fonc.2021.709210

PMID: 34367994
Sample: MG63 cells,U-2 OS cells,Saos-2 cells, 143B cells,K
Frontiers in Pharmacology (2020) IF: 4.225
Corydalis Saxicola Bunting Total Alkaloids Attenuate Walker 256-Induced Bone Pain and Osteoclastogenesis by Suppressing RANKL-Induced NF-κB and c-Fos/NFATc1 Pathways in Rats

DOI: 10.3389/fphar.2020.609119

PMID: 33574755
Sample: serum,cell culture media
CELL BIOLOGY INTERNATIONAL (2023) IF: 4.473
BML-111 inhibits osteoclast differentiation by suppressing the MAPK and NF-κB pathways, alleviating deterioration of the knee joints in a CIA rat model

DOI: 10.1002/cbin.11990

Sample: serum
World Journal of Stem Cells (2020) IF: 3.231
CR6-interacting factor-1 contributes to osteoclastogenesis by inducing receptor activator of nuclear factor κB ligand after radiation

DOI: 10.4252/wjsc.v12.i3.222

Sample: H-BM-MSCs
Journal of Immunology Research (2019) IF: 3.404
Assessment of Salivary Levels of RANKL and OPG in Aggressive versus Chronic Periodontitis

DOI: 10.1155/2019/6195258

Sample: saliva
CYTOKINE (2018) IF: 3.514
Evaluation of gingival crevicular fluid and peri-implant crevicular fluid levels of sclerostin, TWEAK, RANKL and OPG

DOI: 10.1016/j.cyto.2018.10.021

PMID: 30377055
Sample: Gingival Crevicular Fluid
American Journal of Translational Research (2023) IF: 3.94
Effects of joint irrigation combined with ozone injection on bone metabolism, inflammatory factors, and joint function in knee osteoarthritis.

PMID: 36777823
Sample: serum
JOURNAL OF ENDODONTICS (2019) IF: 2.833
Effect of Calcium Hydroxide Alone or in Combination with Ibuprofen and Ciprofloxacin on Nuclear Factor Kappa B Ligand and Osteoprotegerin Level in Periapical Lesions: A Randomized Controlled Clinical Study

DOI: 10.1016/j.joen.2019.09.011

Sample: interstitial fluid
JOURNAL OF PEDIATRIC HEMATOLOGY ONCOLOGY (2022) IF: 1.17
OPG/RANK/RANKL Axis in Egyptian Children With Acute Lymphoblastic Leukemia After Maintenance Therapy: Relationship to Bone Mineral and Vitamin D Status

DOI: 10.1097/MPH.0000000000002603

Sample: serum
International Journal of Lower Extremity Wounds (2019) IF: 1.277
RANKL Gene Polymorphism as a Potential Biomarker to Identify Acute Charcot Foot Among Indian Population With Type 2 Diabetes: A Preliminary Report:

DOI: 10.1177/1534734619859730

Sample: serum
BMC MUSCULOSKELETAL DISORDERS (2018) IF: 1.998
Sera of patients with axial spondyloarthritis (axSpA) enhance osteoclastogenic potential of monocytes isolated from healthy individuals

DOI: 10.1186/s12891-018-2356-4

PMID: 30522483
Sample: Serum
Journal of Oral Science (2017) IF: 0.876
Assessment of local and systemic 25-hydroxy-vitamin D, RANKL, OPG, and TNF levels in patients with rheumatoid arthritis and periodontitis

DOI: 10.2334/josnusd.16-0677

Sample: Gingival crevicular fluid,Serum

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