Mouse CXCL12/SDF-1 DuoSet ELISA

Catalog # Availability Size / Price Qty
DY460
Ancillary Products Available
Mouse CXCL12 / SDF-1 ELISA Standard Curve
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Product Details
Procedure
Citations (20)
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Mouse CXCL12/SDF-1 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Sufficient Materials
For fifteen 96-well plates*
Specificity
Please see the product datasheet

* 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)

 

Data Example

Mouse CXCL12 / SDF-1 ELISA Standard Curve

Product Datasheets

Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

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.

Entrez Gene IDs:
6387 (Human); 20315 (Mouse); 493806 (Feline); 574334 (Rhesus Macaque)
Alternate Names:
chemokine (C-X-C motif) ligand 12; C-X-C motif chemokine 12; CXCL12; hIRH; hSDF-1; Intercrine reduced in hepatomas; IRH; PBSF; PBSFSDF-1a; Pre-B cell growth-stimulating factor; SCYB12; SDF1; SDF-1; SDF1A; SDF1B; SDF1SDF-1b; stromal cell-derived factor 1; TLSF; TLSF-a; TLSF-b; TPAR1

Assay Procedure

GENERAL ELISA PROTOCOL

Plate Preparation

  1. 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.
  2. 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.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. 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.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. 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.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. 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.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. 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.

20 Citations: Showing 1 - 10
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  1. 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;0(0):.
    Species: Mouse
    Sample Types: Tissue Lysates
  2. 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;76(0):33-43.
    Species: Mouse
    Sample Types: Nanocomplex
  3. 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;4(9):.
    Species: Mouse
    Sample Types: Bone Extract
  4. 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;0(0):.
    Species: Mouse
    Sample Types: Serum
  5. 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;0(0):.
    Species: Human
    Sample Types: Tissue Homogenates
  6. 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, 2018;5(1):67-82.e1.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  7. 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;9(0):258.
  8. 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;13(0):7395-7408.
    Species: Human
    Sample Types: Cell Culture Supernates
  9. 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;20(1):40-47.
    Species: Mouse
    Sample Types: Tissue Homogenates
  10. 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;7(0):45991.
    Species: Mouse
    Sample Types: Interstitial Fluid
  11. 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;0(0):.
    Species: Mouse
    Sample Types: Serum
  12. The time-of-day of myocardial infarction onset affects healing through oscillations in cardiac neutrophil recruitment
    EMBO Mol Med, 2016;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  13. SDF-1?-induced dual pairs of E-selectin/ligand mediate endothelial progenitor cell homing to critical ischemia
    Sci Rep, 2016;6(0):34416.
    Species: Human
    Sample Types: Serum
  14. 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;16(8):850-8.
  15. 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;18(5):790-800.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  16. 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, 2014;44(2):600-8.
    Species: Mouse
    Sample Types: Tissue Homogenates
  17. 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;191(11):5684-94.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  18. 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;209(3):551-63.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  19. 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;7(3):e1001321.
    Species: Mouse
    Sample Types: Tissue Homogenates
  20. 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, 2006;38(4):497-508.
    Species: Mouse
    Sample Types: Cell Culture Supernates

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