Human BCMA/TNFRSF17 DuoSet ELISA Summary
* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.
This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human BCMA/TNFRSF17. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.
Product Features
- Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
- Development protocols are provided to guide further assay optimization
- Assay can be customized to your specific needs
- Economical alternative to complete kits
Kit Content
- Capture Antibody
- Detection Antibody
- Recombinant Standard
- Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)
Other Reagents Required
DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.
The components listed above may be purchased separately:
PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered
Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4
Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered
Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)
Stop Solution: 2 N H2SO4 (Catalog # DY994)
Microplates: R&D Systems (Catalog # DY990)
Plate Sealers: ELISA Plate Sealers (Catalog # DY992)
Scientific Data
Product Datasheets
Preparation and Storage
Background: BCMA/TNFRSF17
B cell maturation protein (BCMA) is a member of the TNF receptor superfamily (TNFRSF17). BCMA is expressed in immune organs and mature Bcell lines. Although some expression has been observed at the cell surface, BCMA appears to be localized to the Golgi compartment. BCMA binding to APRIL or BAFF stimulates IgM production in peripheral blood B cells and increases the survival of cultured B cells. Mature human and mouse BCMA share 86% amino acid sequence identity.
Assay Procedure
GENERAL ELISA PROTOCOL
Plate Preparation
- Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
- Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
- Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
- Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.
Assay Procedure
- Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
- Repeat the aspiration/wash as in step 2.
- Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
- Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
- Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.
Citations for Human BCMA/TNFRSF17 DuoSet ELISA
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
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Citations: Showing 1 - 9
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Mechanisms of antigen escape from BCMA- or GPRC5D-targeted immunotherapies in multiple myeloma
Authors: Lee, H;Ahn, S;Maity, R;Leblay, N;Ziccheddu, B;Truger, M;Chojnacka, M;Cirrincione, A;Durante, M;Tilmont, R;Barakat, E;Poorebrahim, M;Sinha, S;McIntyre, J;M Y Chan, A;Wilson, H;Kyman, S;Krishnan, A;Landgren, O;Walter, W;Meggendorfer, M;Haferlach, C;Haferlach, T;Einsele, H;Kortüm, MK;Knop, S;Alberge, JB;Rosenwald, A;Keats, JJ;Rasche, L;Maura, F;Neri, P;Bahlis, NJ;
Nature medicine
Species: Human
Sample Types: Cell Culture Supernates
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A Proliferation-Inducing Ligand and B-Cell Activating Factor Are Upregulated in Patients with Essential Thrombocythemia
Authors: L Bolkun, M Tynecka, T Wasiluk, J Piszcz, A Starosz, K Grubczak, M Moniuszko, A Eljaszewic
Journal of Clinical Medicine, 2022-08-09;11(16):.
Species: Human
Sample Types: Serum
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Serum BCMA levels predict outcomes in MGUS and smoldering myeloma patients
Authors: A Visram, C Soof, SV Rajkumar, SK Kumar, S Bujarski, TM Spektor, RA Kyle, JR Berenson, A Dispenzier
Blood Cancer Journal, 2021-06-24;11(6):120.
Species: Human
Sample Types: Serum
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Characterization of B- and T-Cell Compartment and B-Cell Related Factors Belonging to the TNF/TNFR Superfamily in Patients With Clinically Active Systemic Lupus Erythematosus: Baseline BAFF Serum Levels Are the Strongest Predictor of Response to Belimumab after Twelve Months of Therapy
Authors: S Piantoni, F Regola, S Masneri, M Merletti, T Lowin, P Airò, A Tincani, F Franceschi, L Andreoli, G Pongratz
Frontiers in Pharmacology, 2021-05-21;12(0):666971.
Species: Human
Sample Types: Serum
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Preclinical development of a humanized chimeric antigen receptor against B cell maturation antigen for multiple myeloma
Authors: L Perez-Amil, G Suñe, A Antoñana-V, M Castella, A Najjar, J Bonet, N Fernández-, S Inogés, A López, C Bueno, M Juan, A Urbano-Isp, B Martín-Ant
Haematologica, 2021-01-01;0(0):.
Species: Mouse
Sample Types: Cell Culture Supernates
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Cerebrospinal Fluid Biomarkers in Relation to MRZ Reaction Status in Primary Progressive Multiple Sclerosis
Authors: T Robinson, A Abdelhak, T Bose, E Meinl, M Otto, UK Zettl, R Dersch, H Tumani, S Rauer, A Huss
Cells, 2020-11-25;9(12):.
Species: Human
Sample Types: CSF
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B cell maturation antigen-specific CAR T cells are clinically active in multiple myeloma.
Authors: Cohen A, Garfall A, Stadtmauer E, Melenhorst J, Lacey S, Lancaster E, Vogl D, Weiss B, Dengel K, Nelson A, Plesa G, Chen F, Davis M, Hwang W, Young R, Brogdon J, Isaacs R, Pruteanu-Malinici I, Siegel D, Levine B, June C, Milone M
J Clin Invest, 2019-03-21;130(0):.
Species: Human
Sample Types: Serum (confirm CAR-T)
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Human Plasmacytoid Dendritic Cells Display and Shed B Cell Maturation Antigen upon TLR Engagement
Authors: E Schuh, A Musumeci, FS Thaler, S Laurent, JW Ellwart, R Hohlfeld, A Krug, E Meinl
J. Immunol, 2017-03-10;0(0):.
Species: Human
Sample Types: Cell Lysates
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B-cell reconstitution and BAFF after alemtuzumab (Campath-1H) treatment of multiple sclerosis.
Authors: Thompson SA, Jones JL, Cox AL, Compston DA, Coles AJ
J. Clin. Immunol., 2009-09-10;30(1):99-105.
Species: Human
Sample Types: Serum
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DY193 is a very good assay which has assay dilutional linearity and assay precision.
