Mouse CXCL12/SDF-1 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 mouse CXCL12/SDF-1. 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.
Normal Goat Serum: (Catalog # DY005)
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)
Normal Goat Serum: (Catalog # DY005)
Scientific Data
Product Datasheets
Preparation and Storage
Background: CXCL12/SDF-1
CXCL12/Stromal Cell-derived Factor 1 (SDF-1) is a member of the CXC chemokine ligand superfamily. Six alternatively spliced CXCL12/SDF-1 isoforms have been identified including, CXCL12/SDF-1 alpha, CXCL12/SDF-1 beta, and CXCL12/SDF-1 gamma. CXCL12/SDF-1 alpha is 89 amino acids (aa) in length with a predicted molecular weight of 10 kDa, CXCL12/SDF-1 beta is 93 aa in length with a predicted molecular weight of 10.6 kDa, and CXCL12/SDF-1 gamma is 119 aa in length with a predicted molecular weight of 13.7 kDa. The mouse CXCL12/SDF-1 orthologs share 99% aa sequence identity with the human protein. The alpha and beta isoforms are ubiquitously expressed while the gamma isoform is primarily expressed in the heart. CXCL12/SDF-1 isoforms are chemotactic for T lymphocytes and monocytes. It is cardio protective and has also been shown to hinder CXCR4-mediated HIV infection of T cells.
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 with NGS, 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 Mouse CXCL12/SDF-1 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.
24
Citations: Showing 1 - 10
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HMGB1 promotes CXCL12-dependent egress of murine B cells from Peyer's patches in homeostasis
Authors: L Spagnuolo, V Puddinu, N Boss, T Spinetti, A Oberson, J Widmer, I Mottas, C Hotz, ME Bianchi, M Uguccioni, C Bourquin
European Journal of Immunology, 2021-06-16;0(0):.
Species: Mouse
Sample Types: Cell Culture Supernates
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The longevity-associated variant of BPIFB4 improves a CXCR4-mediated striatum-microglia crosstalk preventing disease progression in a mouse model of Huntington's disease
Authors: A Di Pardo, E Ciaglia, M Cattaneo, A Maciag, F Montella, V Lopardo, A Ferrario, F Villa, M Madonna, E Amico, A Carrizzo, A Damato, G Pepe, F Marracino, A Auricchio, C Vecchione, V Maglione, AA Puca
Cell Death Dis, 2020-07-18;11(7):546.
Species: Mouse
Sample Types: Cell Culture Supernates
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Regulatory T cells confer a circadian signature on inflammatory arthritis
Authors: LE Hand, KJ Gray, SH Dickson, DA Simpkins, DW Ray, JE Konkel, MR Hepworth, JE Gibbs
Nat Commun, 2020-04-03;11(1):1658.
Species: Mouse
Sample Types: Serum
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Functional loss of inactive rhomboid-like protein�2 mitigates obesity by suppressing pro-inflammatory macrophage activation-triggered adipose inflammation
Authors: M Xu, C Ge, Y Qin, D Lou, Q Li, J Feng, Y Wu, L Hu, B Wang, J Tan
Mol Metab, 2020-01-22;34(0):112-123.
Species: Mouse
Sample Types: Cell Culture Supernates
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ATF2 and ATF7 Are Critical Mediators of Intestinal Epithelial Repair
Authors: BJ Meijer, FP Giugliano, B Baan, JHM van der Me, S Meisner, M van Roest, PJ Koelink, RJ de Boer, N Jones, W Breitwiese, NN van der We, ME Wildenberg, GR van den Br, J Heijmans, V Muncan
Cell Mol Gastroenterol Hepatol, 2020-01-17;0(0):.
Species: Mouse
Sample Types: Tissue Lysates
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Decreased viability and neurite length in neural cells treated with chitosan-dextran sulfate nanocomplexes
Authors: LN Zamproni, D Teixeira, A Arnaut All, I Longo Maug, A des Rieux, MA Porcionatt
Neurotoxicology, 2019-10-09;76(0):33-43.
Species: Mouse
Sample Types: Nanocomplex
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Complement receptor C3aR1 controls neutrophil mobilization following spinal cord injury through physiological antagonism of CXCR2
Authors: FH Brennan, T Jogia, ER Gillespie, LV Blomster, XX Li, B Nowlan, GM Williams, E Jacobson, GW Osborne, FA Meunier, SM Taylor, KE Campbell, KP MacDonald, JP Levesque, TM Woodruff, MJ Ruitenberg
JCI Insight, 2019-05-02;4(9):.
Species: Mouse
Sample Types: Bone Extract
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ATF3 and JDP2 deficiency in cancer associated fibroblasts promotes tumor growth via SDF-1 transcription
Authors: S Avraham, B Korin, S Aviram, D Shechter, Y Shaked, A Aronheim
Oncogene, 2019-01-22;0(0):.
Species: Mouse
Sample Types: Serum
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Nanoparticle-modified chitosan-agarose-gelatin scaffold for sustained release of SDF-1 and BMP-2
Authors: B Wang, Y Guo, X Chen, C Zeng, Q Hu, W Yin, W Li, H Xie, B Zhang, X Huang, F Yu
Int J Nanomedicine, 2018-11-12;13(0):7395-7408.
Species: Human
Sample Types: Cell Culture Supernates
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A Novel Role for C5a in B-1 Cell Homeostasis
Authors: K Bröker, J Figge, AF Magnusen, RA Manz, J Köhl, CM Karsten
Front Immunol, 2018-02-19;9(0):258.
Species: Mouse
Sample Types: Peritoneal Lavage Fluid
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Expression of the B cell differentiation factor BAFF and chemokine CXCL13 in a murine model of Respiratory Syncytial Virus infection
Authors: W Alturaiki, AJ McFarlane, K Rose, R Corkhill, PS McNamara, J Schwarze, BF Flanagan
Cytokine, 2018-02-01;0(0):.
Species: Human
Sample Types: Tissue Homogenates
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Indian Hedgehog Suppresses a Stromal Cell-Driven Intestinal Immune Response
Authors: BF Westendorp, NVJA Büller, ON Karpus, WA van Dop, J Koster, R Versteeg, PJ Koelink, CY Snel, S Meisner, JJTH Roelofs, A Uhmann, E Ver Loren, J Heijmans, H Hahn, V Muncan, ME Wildenberg, GR van den Br
Cell Mol Gastroenterol Hepatol, 2017-09-05;5(1):67-82.e1.
Species: Mouse
Sample Types: Cell Culture Supernates
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MCPIP1 downregulation in clear cell renal cell carcinoma promotes vascularization and metastatic progression
Authors: P Marona, J Górka, Z Mazurek, W Wilk, J Rys, M Majka, J Jura, K Miekus
Cancer Res., 2017-07-17;0(0):.
Species: Mouse
Sample Types: Serum
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Stromal Cell-Derived Factor 1 Mediates Immune Cell Attraction upon Urinary Tract Infection
Authors: B Isaacson, T Hadad, A Glasner, C Gur, Z Granot, G Bachrach, O Mandelboim
Cell Rep, 2017-07-05;20(1):40-47.
Species: Mouse
Sample Types: Tissue Homogenates
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Protein malnutrition promotes dysregulation of molecules involved in T cell migration in the thymus of mice infected with Leishmania infantum
Authors: M Losada-Bar, A Umaña-Pére, S Cuervo-Esc, LR Berbert, R Porrozzi, FN Morgado, DA Mendes-da-, W Savino, M Sánchez-Gó, P Cuervo
Sci Rep, 2017-04-11;7(0):45991.
Species: Mouse
Sample Types: Interstitial Fluid
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SDF-1?-induced dual pairs of E-selectin/ligand mediate endothelial progenitor cell homing to critical ischemia
Sci Rep, 2016-10-07;6(0):34416.
Species: Human
Sample Types: Serum
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The time-of-day of myocardial infarction onset affects healing through oscillations in cardiac neutrophil recruitment
EMBO Mol Med, 2016-08-01;0(0):.
Species: Mouse
Sample Types: Cell Culture Supernates
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Dipeptidylpeptidase 4 inhibition enhances lymphocyte trafficking, improving both naturally occurring tumor immunity and immunotherapy.
Authors: Barreira da Silva R, Laird M, Yatim N, Fiette L, Ingersoll M, Albert M
Nat Immunol, 2015-06-15;16(8):850-8.
Species: Mouse
Sample Types: Tissue Homogenates
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Controlled intramyocardial release of engineered chemokines by biodegradable hydrogels as a treatment approach of myocardial infarction.
Authors: Projahn D, Simsekyilmaz S, Singh S, Kanzler I, Kramp B, Langer M, Burlacu A, Bernhagen J, Klee D, Zernecke A, Hackeng T, Groll J, Weber C, Liehn E, Koenen R
J Cell Mol Med, 2014-02-06;18(5):790-800.
Species: Mouse
Sample Types: Cell Culture Supernates
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Molecular changes in bone marrow, tumor and serum after conductive ablation of murine 4T1 breast carcinoma.
Authors: Przybyla B, Shafirstein G, Vishal S, Dennis R, Griffin R
Int J Oncol, 2013-11-21;44(2):600-8.
Species: Mouse
Sample Types: Tissue Homogenates
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CX3CR1 regulates the maintenance of KLRG1+ NK cells into the bone marrow by promoting their entry into circulation.
Authors: Ponzetta A, Sciume G, Benigni G, Antonangeli F, Morrone S, Santoni A, Bernardini G
J Immunol, 2013-11-01;191(11):5684-94.
Species: Mouse
Sample Types: Cell Culture Supernates
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HMGB1 promotes recruitment of inflammatory cells to damaged tissues by forming a complex with CXCL12 and signaling via CXCR4.
Authors: Schiraldi M, Raucci A, Munoz LM, Livoti E, Celona B, Venereau E, Apuzzo T, De Marchis F, Pedotti M, Bachi A, Thelen M, Varani L, Mellado M, Proudfoot A, Bianchi ME, Uguccioni M
J. Exp. Med., 2012-02-27;209(3):551-63.
Species: Mouse
Sample Types: Cell Culture Supernates
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Chemokine binding protein M3 of murine gammaherpesvirus 68 modulates the host response to infection in a natural host.
Authors: Hughes DJ, Kipar A, Leeming GH, Bennett E, Howarth D, Cummerson JA, Papoula-Pereira R, Flanagan BF, Sample JT, Stewart JP
PLoS Pathog., 2011-03-17;7(3):e1001321.
Species: Mouse
Sample Types: Tissue Homogenates
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Regulation of SDF-1 (CXCL12) production by osteoblasts; a possible mechanism for stem cell homing.
Authors: Jung Y, Wang J, Schneider A, Sun YX, Koh-Paige AJ, Osman NI, McCauley LK, Taichman RS
Bone, 2005-12-05;38(4):497-508.
Species: Mouse
Sample Types: Cell Culture Supernates
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