CXCL2 ELISA Kit (Chemokine (C-X-C Motif) Ligand 2)

Details for Product CXCL2 ELISA Kit No. ABIN1979727, Supplier: Log in to see
Antigen
  • CINC-2a
  • GRO2
  • GROb
  • MGSA-b
  • MIP-2a
  • MIP2
  • MIP2A
  • SCYB2
  • GRO3
  • Gro2
  • MIP-2
  • Mgsa-b
  • Mip2
  • Scyb
  • Scyb2
  • Mip-2
  • GROB
  • MIP-2alpha
  • C-X-C motif chemokine ligand 2
  • chemokine (C-X-C motif) ligand 2
  • CXCL2
  • Cxcl2
Reactivity
Mouse (Murine)
Alternatives
Kits with alternative reactivity to:
37
33
28
3
3
3
3
2
2
2
1
1
Method Type
Sandwich ELISA
Detection Range
10-2000 pg/mL
Minimum Detection Limit
10 pg/mL
Application
ELISA
Options
Supplier
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Purpose Mouse MIP-2 ELISA Kit for cell culture supernatants, plasma, and serum samples.
Sample Type Serum, Plasma, Cell Culture Supernatant
Analytical Method Quantitative
Detection Method Colorimetric
Specificity This ELISA kit shows no cross-reactivity with any of the cytokines tested: Mouse CD30, L CD30, T CD40, CRG-2, CTACK, CXCL16, Eotaxin , Eotaxin-2, Fas Ligand, Fractalkine, GCSF, GM-CFS, IFN- gamma, IGFBP-3, IGFBP-5, IGFBP-6, IL-1 alpha, IL-1 beta, IL-2, IL-3, IL-3 Rb, IL-4, IL-5, IL-6, IL-9, IL-10, IL-12 p40/p70, IL-12 p70, IL-13, IL-17, KC, Leptin R, LEPTIN(OB), LIX, L-Selectin, Lymphotactin, MCP-1, MCP-5, M-CSF, MIG, MIP-1 alpha, MIP-1 gamma, MIP-3 beta, MIP-3 alpha, PF-4, P-Selectin, RANTES, SCF, SDF-1 alpha, TARC, TCA-3, TECK, TIMP-1, TNF-alpha, TNF RI, TNF RII, TPO, VCAM-1, VEGF.
Sensitivity < 10 pg/mL
Characteristics
  • Strip plates and additional reagents allow for use in multiple experiments
  • Quantitative protein detection
  • Establishes normal range
  • The best products for confirmation of antibody array data
Components
  • Pre-Coated 96-well Strip Microplate
  • Wash Buffer
  • Stop Solution
  • Assay Diluent(s)
  • Lyophilized Standard
  • Biotinylated Detection Antibody
  • Streptavidin-Conjugated HRP
  • TMB One-Step Substrate
Material not included
  • Distilled or deionized water
  • Precision pipettes to deliver 2 μL to 1 μL volumes
  • Adjustable 1-25 μL pipettes for reagent preparation
  • 100 μL and 1 liter graduated cylinders
  • Tubes to prepare standard and sample dilutions
  • Absorbent paper
  • Microplate reader capable of measuring absorbance at 450nm
  • Log-log graph paper or computer and software for ELISA data analysis
Alternative Name MIP-2 / CXCL2 (CXCL2 ELISA Kit Abstract)
Background MIP-2 (macrophage inflammatory protein) is a basic protein of approximately 6 kDa. MIP-2 is extremely chemotactic for segmented neutrophilic granulocytes and also synergises with GM-CSF and M-CSF. It also induces the degranulation of human neutrophils but it does not enhance oxidative metabolism. Mouse MIP-2 exhibits potent neutrophil chemotactic activity and may be a key mediator of neutrophil recruitment in response to tissue injury and infection. The Mouse MIP-2 ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of mouse MIP-2 in serum, plasma and cell culture supernatants. This assay employs an antibody specific for mouse MIP-2 coated on a 96-well plate. Standards and samples are pipetted into the wells and MIP-2 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-mouse MIP-2 antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of MIP-2 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm. Reproducibility: Intra-Assay: CV<10% Inter-Assay: CV<12%.
Gene ID 20310
UniProt P10889
Pathways Cellular Response to Molecule of Bacterial Origin
Application Notes Recommended Dilution for serum and plasma samples2 fold
Sample Volume 100 μL
Plate Pre-coated
Protocol
  1. Prepare all reagents, samples and standards as instructed in the manual.
  2. Add 100 μL of standard or sample to each well.
  3. Incubate 2.5 h at RT or O/N at 4 °C.
  4. Add 100 μL of prepared biotin antibody to each well.
  5. Incubate 1 h at RT.
  6. Add 100 μL of prepared Streptavidin solution to each well.
  7. Incubate 45 min at RT.
  8. Add 100 μL of TMB One-Step Substrate Reagent to each well.
  9. Incubate 30 min at RT.
  10. Add 50 μL of Stop Solution to each well.
  11. Read at 450 nm immediately.
Reagent Preparation
  1. Bring all reagents and samples to room temperature (18 - 25 °C) before use.
    2. Sample dilution: If your samples need to be diluted, Assay Diluent A (Item D) should be used for dilution of serum/plasma samples. 1x Assay Diluent B (Item E) should be used for dilution of culture supernatants. Suggested dilution for normal serum/plasma: 2 fold. Please note that levels of the target protein may vary between different specimens. Optimal dilution factors for each sample must be determined by the investigator.
    3. Assay Diluent B should be diluted 5-folds with deionized or distilled water.
    4. Preparation of standard: Briefly spin the vial of Item C and then add 400 µL Assay Diluent A (for serum/plasma samples) or 1x Assay Diluent B (for cell culture medium) into Item C vial to prepare a 50 ng/mL standard. Dissolve the powder thoroughly by a gentle mix. Add 40 µL MIP-2 standard from the vial of Item C, into a tube with 960.0 µL Assay Diluent A or 1x Assay Diluent B to prepare a 2000 pg/mL stock standard solution. Pipette 400 µL Assay Diluent A or 1x Assay Diluent B into each tube. Use the stock standard solution to produce a dilution series . Mix each tube thoroughly before the next transfer. Assay Diluent A or 1x Assay Diluent B serves as the zero standard (0 pg/mL). 200 µL 200 µL 200 µL 200 µL 200 µL 200myl 40 µL standard + 960 µL 2000 666.7 222.2 74.07 24.69 8.23 2.74 0 pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL
    5. If the Wash Concentrate (20x) (Item B) contains visible crystals, warm to room temperature and mix gently until dissolved. Dilute 20 ml of Wash Buffer Concentrate into deionized or distilled water to yield 400 ml of 1x Wash Buffer.
    6. Briefly spin the Detection Antibody vial (Item F) before use. Add 100 µL of 1x Assay Diluent B into the vial to prepare a detection antibody concentrate. Pipette up and down to mix gently (the concentrate can be stored at 4 °C for 5 days). The detection antibody concentrate should be diluted 90-folds with 1x Assay Diluent B and used in step 4 of Part VI Assay Procedure.
    7. Briefly spin the HRP-Streptavidin concentrate vial (Item G) and pipette up and down to mix gently before use. HRP-Streptavidin concentrate should be diluted 700-fold with 1x Assay Diluent B. For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 20 µL of HRP-Streptavidin concentrate into a tube with 14 ml 1x Assay Diluent B to prepare a 700-fold diluted HRP-Streptavidin solution (don't store the diluted solution for next day use). Mix well.
Assay Procedure
  1. Bring all reagents and samples to room temperature (18 - 25 °C) before use. It is recommended that all standards and samples be run at least in duplicate.
    2. Add 100 µL of each standard (see Reagent Preparation step 2) and sample into appropriate wells. Cover well and incubate for 2.5 hours at room temperature or over night at 4 °C with gentle shaking.
    3. Discard the solution and wash 4 times with 1x Wash Solution. Wash by filling each well with Wash Buffer (300 myl) using a multi-channel Pipette or autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels.
    4. Add 100 µL of 1x prepared biotinylated antibody (Reagent Preparation step 6) to each well. Incubate for 1 hour at room temperature with gentle shaking.
    5. Discard the solution. Repeat the wash as in step
    6. Add 100 µL of prepared Streptavidin solution (see Reagent Preparation step 7) to each well. Incubate for 45 minutes at room temperature with gentle shaking.
    7. Discard the solution. Repeat the wash as in step
    8. Add 100 µL of TMB One-Step Substrate Reagent (Item H) to each well. Incubate for 30 minutes at room temperature in the dark with gentle shaking.
    9. Add 50 µL of Stop Solution (Item I) to each well. Read at 450 nm immediately.
Calculation of Results

Calculate the mean absorbance for each set of duplicate standards, controls and samples, and subtract the average zero standard optical density. Plot the standard curve on log-log graph paper or using Sigma plot software, with standard concentration on the x-axis and absorbance on the y-axis. Draw the best-fit straight line through the standard points.
Typical Data: These standard curves are for demonstration only. A standard curve must be run with each assay. Assay Diluent A Mouse MIP-2 concentration (pg/mL) 1 10 100 1000 10000 O D =4 50 n m 0.01 0.1 1 10 Assay Diluent B Mouse MIP-2 concentration (pg/mL) 1 10 100 1000 10000 O D =4 50 n m 0.001 0.01 0.1 1 10
Sensitivity: The minimum detectable dose of MIP-2 is typically less than 10 pg/mL.
Recovery: Recovery was determined by spiking various levels of mouse MIP-2 into mouse serum, plasma and cell culture media. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Serum 95.34 82-103 Plasma 92.63 80-102 Cell culture media 98.16 87-108
Linearity: Sample Type Serum Plasma Cell Culture Media 1:2 Average % of Expected 94 95 98 Range ( %) 82-103 83-103 84-105 1:4 Average % of Expected 96 93 101 Range ( %) 83-104 81-102 88-108
Reproducibility: Intra-Assay: CV<10 % Inter-Assay: CV<12 %

Assay Precision Intra-Assay: CV< 10 % Inter-Assay: CV< 12 %
Restrictions For Research Use only
Handling Advice Avoid repeated freeze-thaw cycles.
Storage -20 °C
Storage Comment The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.
Expiry Date 6 months
Supplier Images
ELISA image for Chemokine (C-X-C Motif) Ligand 2 (CXCL2) ELISA Kit (ABIN1979727) Chemokine (C-X-C Motif) Ligand 2 (CXCL2) ELISA Kit
Product cited in: Shanmugam, Wang, Markand, Perry, Tawfik, Zorrilla, Ganapathy, Smith: "Sigma receptor 1 activation attenuates release of inflammatory cytokines MIP1?, MIP2, MIP3?, and IL12 (p40/p70) by retinal Müller glial cells." in: Journal of neurochemistry, 2015 (PubMed).

Kumar, Sahu, Kumar, Subuddhi, Maji, Jana, Gupta, Raffetseder, Lerm, Ghosh, van Loo, Beyaert, Gupta, Kundu, Basu: "MicroRNA let-7 modulates the immune response to Mycobacterium tuberculosis infection via control of A20, an inhibitor of the NF-?B pathway." in: Cell host & microbe, Vol. 17, Issue 3, pp. 345-56, 2015 (PubMed).

Evans, Killoran, Mitre, Morris, Kim, Mitre: "Ten Weeks of Infection with a Tissue-Invasive Helminth Protects against Local Immune Complex-Mediated Inflammation, but Not Cutaneous Type I Hypersensitivity, in Previously Sensitized Mice." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 195, Issue 7, pp. 2973-84, 2015 (PubMed).

Xu, Shao, Wang, Zhou, Tang, Lu, Xiong: "Role of Interleukin-17 in defense against pseudomonas aeruginosa infection in lungs." in: International journal of clinical and experimental medicine, Vol. 7, Issue 4, pp. 809-16, 2014 (PubMed).

Liu, Petersen, Devireddy: "Impaired neutrophil function in 24p3 null mice contributes to enhanced susceptibility to bacterial infections." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 190, Issue 9, pp. 4692-706, 2013 (PubMed).

Said, Frierson, Sanchez-Carbayo, Brekken, Theodorescu: "Loss of SPARC in bladder cancer enhances carcinogenesis and progression." in: The Journal of clinical investigation, Vol. 123, Issue 2, pp. 751-66, 2013 (PubMed).

Sasakawa, Kominami, Yamamoto, Nakaoka, Nakamura, Nakao, Abe, Fukuhama, Kagawa: "Effects of globin digest and its active ingredient Trp-Thr-Gln-Arg on galactosamine/lipopolysaccharide-induced liver injury in ICR mice." in: Life sciences, Vol. 90, Issue 5-6, pp. 190-9, 2012 (PubMed).

Oh, Woo, Lim, Moon: "ERK2-dependent activation of c-Jun is required for nontypeable Haemophilus influenzae-induced CXCL2 upregulation in inner ear fibrocytes." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 188, Issue 7, pp. 3496-505, 2012 (PubMed).

Sherry, Horii, Manogue, Widmer, Cerami: "Macrophage inflammatory proteins 1 and 2: an overview." in: Cytokines, Vol. 4, pp. 117-30, 1993

Background publications Wolpe, Cerami: "Macrophage inflammatory proteins 1 and 2: members of a novel superfamily of cytokines." in: FASEB journal : official publication of the Federation of American Societies for Experimental Biology, Vol. 3, Issue 14, pp. 2565-73, 1990 (PubMed).

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