Human 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 human 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 Human 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.
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Citations: Showing 1 - 10
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Baseline circulating biomarkers, their changes, and subsequent suicidal ideation and depression severity at 6 months: A prospective analysis in patients with mood disorders
Authors: Lengvenyte, A;Cognasse, F;Hamzeh-Cognasse, H;Sénèque, M;Strumila, R;Olié, E;Courtet, P;
Psychoneuroendocrinology
Species: Human
Sample Types: Plasma
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Unraveling the impact of cancer-associated fibroblasts on hypovascular pancreatic neuroendocrine tumors
Authors: Lai, TY;Chiang, TC;Lee, CY;Kuo, TC;Wu, CH;Chen, YI;Hu, CM;Maskey, M;Tang, SC;Jeng, YM;Tien, YW;Lee, EYP;Lee, WH;
British journal of cancer
Species: Human
Sample Types: Cell Culture Supernates
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The long non-coding RNA HOTAIR contributes to joint-specific gene expression in rheumatoid arthritis
Authors: Elhai, M;Micheroli, R;Houtman, M;Mirrahimi, M;Moser, L;Pauli, C;Bürki, K;Laimbacher, A;Kania, G;Klein, K;Schätzle, P;Bertoncelj, MF;Edalat, SG;Keusch, L;Khmelevskaya, A;Toitou, M;Geiss, C;Rauer, T;Sakkou, M;Kollias, G;Armaka, M;Distler, O;Ospelt, C;
Nature communications
Species: Human
Sample Types: Cell Culture Supernates
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Decreased Serum Stromal Cell-Derived Factor-1 in Patients with Familial Hypercholesterolemia and Its Strong Correlation with Lipoprotein Subfractions
Authors: Juhász, L;L?rincz, H;Szentpéteri, A;Tóth, N;Varga, É;Paragh, G;Harangi, M;
International journal of molecular sciences
Species: Human
Sample Types: Serum
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Prognostic significance of plasma SDF-1 in acute ischemic stroke patients with diabetes mellitus: the CATIS trial
Authors: You, S;Chen, H;Miao, M;Du, J;Che, B;Xu, T;Liu, CF;Zhang, Y;He, J;Zhong, X;Cao, Y;Zhong, C;
Cardiovascular diabetology
Species: Human
Sample Types: Plasma
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CXCL12 and CXCR4 as Novel Biomarkers in Uric Acid-Induced Inflammation and Patients with Gouty Arthritis
Authors: SK Kim, JY Choe, KY Park
Biomedicines, 2023-02-21;11(3):.
Species: Human
Sample Types: Serum
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Association between the expression of Toll-like receptors, cytokines, and homeostatic chemokines in SARS-CoV-2 infection and COVID-19 severity
Authors: W Alturaiki, H Alkadi, S Alamri, ME Awadalla, A Alfaez, A Mubarak, MA Alanazi, FQ Alenzi, BF Flanagan, B Alosaimi
Heliyon, 2022-12-24;0(0):e12653.
Species: Human
Sample Types: Serum
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The fatal trajectory of pulmonary COVID-19 is driven by lobular ischemia and fibrotic remodelling
Authors: M Ackermann, JC Kamp, C Werlein, CL Walsh, H Stark, V Prade, R Surabattul, WL Wagner, C Disney, AJ Bodey, T Illig, DJ Leeming, MA Karsdal, A Tzankov, P Boor, MP Kühnel, FP Länger, SE Verleden, HM Kvasnicka, HH Kreipe, A Haverich, SM Black, A Walch, P Tafforeau, PD Lee, MM Hoeper, T Welte, B Seeliger, S David, D Schuppan, SJ Mentzer, DD Jonigk
EBioMedicine, 2022-10-04;85(0):104296.
Species: Human
Sample Types: Plasma
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Pro-cancerogenic effects of spontaneous and drug-induced senescence of ovarian cancer cells in vitro and in vivo: a comparative analysis
Authors: S Rutecki, P Szulc, M Paku?a, P Uruski, A Radziemski, E Naumowicz, R Moszy?ski, A Tykarski, J Miku?a-Pie, K Ksi??ek
Journal of ovarian research, 2022-07-26;15(1):87.
Species: Human
Sample Types: Cell Culture Supernates
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Evidence of Stem Cells Mobilization in the Blood of Patients with Pancreatitis: A Potential Link with Disease Severity
Authors: K D?bkowski, A ?ab?d?-Mas, B Do??gowska, K Safranow, M Budkowska, E Zuba-Surma, T Starzy?ska
Stem Cells International, 2022-07-08;2022(0):5395248.
Species: Human
Sample Types: Serum
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MIF/CXCR4 signaling axis contributes to survival, invasion, and drug resistance of metastatic neuroblastoma cells in the bone marrow microenvironment
Authors: L Garcia-Ger, M García, A Garrido-Ga, S Gómez-Gonz, M Torrebadel, E Prada, G Pascual-Pa, O Muñoz, S Perez-Jaum, I Lemos, N Salvador, M Vila-Ubach, A Doncel-Req, M Suñol, AM Carcaboso, J Mora, C Lavarino
BMC Cancer, 2022-06-17;22(1):669.
Species: Human
Sample Types: Cell Culture Supernates
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CHMP2A regulates tumor sensitivity to natural killer cell-mediated cytotoxicity
Authors: D Bernareggi, Q Xie, BC Prager, J Yun, LS Cruz, TV Pham, W Kim, X Lee, M Coffey, C Zalfa, P Azmoon, H Zhu, P Tamayo, JN Rich, DS Kaufman
Nature Communications, 2022-04-07;13(1):1899.
Species: Human
Sample Types: Cell Culture Supernates
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Engagement of the CXCL12-CXCR4 Axis in the Interaction of Endothelial Progenitor Cell and Smooth Muscle Cell to Promote Phenotype Control and Guard Vascular Homeostasis
Authors: SF Mause, E Ritzel, A Deck, F Vogt, EA Liehn
International Journal of Molecular Sciences, 2022-01-14;23(2):.
Species: Human
Sample Types: Cell Culture Supernates
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A Crosslinked Dextran Sulfate-Chitosan Nanoparticle for Delivery of Therapeutic Heparin-binding Proteins
Authors: VA Guarino, A Blau, J Alvarenga, J Loscalzo, YY Zhang
International journal of pharmaceutics, 2021-11-11;0(0):121287.
Species: Rat
Sample Types: Tissue Homogenates
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High circulating SDF-1and MCP-1 levels and genetic variations in CXCL12, CCL2 and CCR5: Prognostic signature of immune recovery status in treated HIV-positive patients
Authors: E Yeregui, C Viladés, P Domingo, A Ceausu, YM Pacheco, S Veloso, A Inciarte, J Vidal-Gonz, M Peraire, C Perpiñán, V Falcó, J Masip, V Alba, M Vargas, A Martí, L Reverté, J Mallolas, F Vidal, J Peraire, A Rull
EBioMedicine, 2020-11-06;62(0):103077.
Species: Human
Sample Types: Plasma
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Clinicogenomic factors of biotherapy immunogenicity in autoimmune disease: A prospective multicohort study of the ABIRISK consortium
Authors: S Hässler, D Bachelet, J Duhaze, N Szely, A Gleizes, S Hacein-Bey, O Aktas, M Auer, J Avouac, M Birchler, Y Bouhnik, O Brocq, D Buck-Marti, G Cadiot, F Carbonnel, Y Chowers, M Comabella, T Derfuss, N De Vries, N Donnellan, A Doukani, M Guger, HP Hartung, E Kubala Hav, B Hemmer, T Huizinga, K Ingenhoven, PE Hyldgaard-, EC Jury, M Khalil, B Kieseier, A Laurén, R Lindberg, A Loercher, E Maggi, J Manson, C Mauri, B Mohand Oum, X Montalban, M Nachury, P Nytrova, C Richez, M Ryner, F Sellebjerg, C Sievers, D Sikkema, M Soubrier, S Tourdot, C Trang, A Vultaggio, C Warnke, S Spindeldre, P Dönnes, TP Hickling, A Hincelin M, M Allez, F Deisenhamm, A Fogdell-Ha, X Mariette, M Pallardy, P Broët, ABIRISK co
PLoS Med, 2020-10-30;17(10):e1003348.
Species: Human
Sample Types: Serum
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Regional hyperthermia enhances mesenchymal stem cell recruitment to tumor stroma: Implications for mesenchymal stem cell-based tumor therapy
Authors: M Tutter, C Schug, KA Schmohl, S Urnauer, C Kitzberger, N Schwenk, M Petrini, C Zach, S Ziegler, P Bartenstei, WA Weber, G Multhoff, E Wagner, LH Lindner, PJ Nelson, C Spitzweg
Mol Ther, 2020-10-15;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
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Effect of Enzymatically Extracted Fucoidans on Angiogenesis and Osteogenesis in Primary Cell Culture Systems Mimicking Bone Tissue Environment
Authors: J Ohmes, Y Xiao, F Wang, MD Mikkelsen, TT Nguyen, H Schmidt, A Seekamp, AS Meyer, S Fuchs
Mar Drugs, 2020-09-21;18(9):.
Species: Human
Sample Types: Cell Culture Supernates
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Development of a Sustained Release Nano-In-Gel Delivery System for the Chemotactic and Angiogenic Growth Factor Stromal-Derived Factor 1alpha
Authors: J O'Dwyer, M Cullen, S Fattah, R Murphy, S Stefanovic, L Kovarova, M Pravda, V Velebny, A Heise, GP Duffy, SA Cryan
Pharmaceutics, 2020-06-04;12(6):.
Species: Human
Sample Types: Protein
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Multiple myeloma exploits Jagged1 and Jagged2 to promote intrinsic and bone marrow-dependent drug resistance
Authors: M Colombo, S Garavelli, M Mazzola, N Platonova, D Giannandre, R Colella, L Apicella, M Lancellott, E Lesma, S Ancona, MT Palano, M Barbieri, E Taiana, E Lazzari, A Basile, M Turrini, A Pistocchi, A Neri, R Chiaramont
Haematologica, 2019-10-03;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
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A Unique Pattern of Mesothelial-Mesenchymal Transition Induced in the Normal Peritoneal Mesothelium by High-Grade Serous Ovarian Cancer
Authors: M Paku?a, P Uruski, A Niklas, A Wo?niak, D Szpurek, A Tykarski, J Miku?a-Pie, K Ksi??ek
Cancers (Basel), 2019-05-13;11(5):.
Species: Human
Sample Types: Cell Culture Supernates
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Fibrin Glue Enhances Adipose-Derived Stromal Cell Cytokine Secretion and Survival Conferring Accelerated Diabetic Wound Healing
Authors: U Hopfner, MM Aitzetmuel, P Ne beta bach, MS Hu, HG Machens, ZN Maan, D Duscher
Stem Cells Int, 2018-12-19;2018(0):1353085.
Species: Human
Sample Types: Cell Culture Supernates
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Functionally distinct disease-associated fibroblast subsets in rheumatoid arthritis
Authors: F Mizoguchi, K Slowikowsk, K Wei, JL Marshall, DA Rao, SK Chang, HN Nguyen, EH Noss, JD Turner, BE Earp, PE Blazar, J Wright, BP Simmons, LT Donlin, GD Kalliolias, SM Goodman, VP Bykerk, LB Ivashkiv, JA Lederer, N Hacohen, PA Nigrovic, A Filer, CD Buckley, S Raychaudhu, MB Brenner
Nat Commun, 2018-02-23;9(1):789.
Species: Human
Sample Types: Cell Culture Supernates
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Serum SDF-1 levels are a reliable diagnostic marker of feline mammary carcinoma, discriminating HER2-overexpressing tumors from other subtypes
Authors: CS Marques, M Soares, A Santos, J Correia, F Ferreira
Oncotarget, 2017-11-11;8(62):105775-105789.
Species: Feline
Sample Types: Serum
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The Role of Shed PrP(c) in the Neuropathogenesis of HIV Infection
Authors: BW Megra, EA Eugenin, JW Berman
J. Immunol., 2017-05-22;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
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MODELING STROMA-INDUCED DRUG RESISTANCE IN A TISSUE-ENGINEERED TUMOR MODEL OF EWING SARCOMA
Authors: Marco Santoro
Tissue Eng Part A, 2017-01-01;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
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Synergic effects of VEGF-A and SDF-1 on the angiogenic properties of endothelial progenitor cells
Authors: Gabriela Odent Grig
J Tissue Eng Regen Med, 2016-12-12;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
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Incorporation of heparin-binding proteins into preformed dextran sulfate-chitosan nanoparticles
Int J Nanomedicine, 2016-11-18;11(0):6149-6159.
Species: Rat
Sample Types: Tissue Homogenates
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Reduction of maternal circulating endothelial progenitor cells in human pregnancies with intrauterine growth restriction.
Authors: Calcaterra F, Taddeo A, Colombo E, Cappelletti M, Martinelli A, Calabrese S, Mavilio D, Cetin I, Della Bella S
Placenta, 2014-04-26;35(7):431-6.
Species: Human
Sample Types: Plasma
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CXCR4 positive and angiogenic monocytes in myocardial infarction.
Thromb Haemost, 2012-12-06;109(2):255-62.
Species: Human
Sample Types: Plasma
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The EGFR ligands amphiregulin and heparin-binding egf-like growth factor promote peritoneal carcinomatosis in CXCR4-expressing gastric cancer.
Authors: Yasumoto K, Yamada T, Kawashima A, Wang W, Li Q, Donev IS, Tacheuchi S, Mouri H, Yamashita K, Ohtsubo K, Yano S
Clin. Cancer Res., 2011-04-11;17(11):3619-30.
Species: Human
Sample Types: Complex Sample Type
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Identification of a New Phenotype of Tolerogenic Human Dendritic Cells Induced by Fungal Proteases from Aspergillus oryzae.
Authors: Zimmer A, Luce S, Gaignier F, Nony E, Naveau M, Biola-Vidamment A, Pallardy M, Van Overtvelt L, Mascarell L, Moingeon P
J. Immunol., 2011-03-02;186(7):3966-76.
Species: Human
Sample Types: Cell Culture Supernates
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Distinct platelet packaging, release, and surface expression of proangiogenic and antiangiogenic factors on different platelet stimuli.
Authors: Chatterjee M, Huang Z, Zhang W, Jiang L, Hultenby K, Zhu L, Hu H, Nilsson GP, Li N
Blood, 2011-02-17;117(14):3907-11.
Species: Human
Sample Types: Cell Culture Supernates
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CXCL12 is essential for migration of activated Langerhans cells from epidermis to dermis.
Authors: Ouwehand K, Santegoets SJ, Bruynzeel DP, Scheper RJ, de Gruijl TD, Gibbs S
Eur. J. Immunol., 2008-11-01;38(11):3050-9.
Species: Human
Sample Types: Cell Culture Supernates
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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: Human
Sample Types: Cell Culture Supernates
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works well for human serum and plasma samples