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

Details for Product CXCL2 ELISA Kit No. ABIN1980083, 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
Rat (Rattus)
Alternatives
Kits with alternative reactivity to:
37
33
28
3
3
3
3
2
2
2
1
1
Method Type
Sandwich ELISA
Detection Range
2-1800 pg/mL
Minimum Detection Limit
2 pg/mL
Application
ELISA
Options
Supplier
Log in to see
Supplier Product No.
Log in to see
Purpose Rat CINC-3 ELISA Kit for cell culture supernatants, plasma, and serum samples.
Sample Type Plasma, Cell Culture Supernatant, Serum
Analytical Method Quantitative
Detection Method Colorimetric
Specificity This ELISA kit shows no cross-reactivity with the following cytokines tested: rat CINC-2, CNTF, Fractalkine, IL-1 alpha, IL-1 beta, IL-4, IL-6, IL-10, GM-CSF, IFN-gamma, Leptin, Lix, MCP-1, MIP-3 alpha, beta-NGF, TIMP-1, TNF-alpha.
Sensitivity < 2 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 CINC-3 (CXCL2 ELISA Kit Abstract)
Background C-X-C motif chemokine 2 (Cytokine-induced neutrophil chemoattractant 3) (CINC-3) (Macrophage inflammatory protein 2) (MIP2)
Gene ID 114105
UniProt P30348
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 C (Item L) should be used for dilution of serum/plasma samples. 1x Assay Diluent B (Item E) should be used for dilution of cell culture supernates and urine. 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-fold with deionized or distilled water before use.
    4. Preparation of standard: Briefly spin the vial of Item C and then add 400 µL Assay Diluent C (for serum/plasma samples) or 1x Assay Diluent B (for cell culture supernates) into Item C vial to prepare a 50 ng/mL standard. Dissolve the powder thoroughly by a gentle mix. Add 18 µL Adiponectin standard (50 ng/mL) from the vial of Item C, into a tube with 482 µL Assay Diluent A or Assay Diluent C to prepare a 1,800 pg/mL stock standard solution. Pipette 300 µL Assay Diluent C or 1x Assay Diluent B into each tube. Use the 1,800 pg/mL standard solution to produce a dilution series . Mix each tube thoroughly before the next transfer. Assay Diluent C or 1x Assay Diluent B serves as the zero standard (0 pg/mL). The 1,800 pg/mL standard in Assay Diluent B may be saturated, we recommend starting from 720 pg/mL. 200 µL 200 µL 200 µL 200 µL 200 µL 200myl 18 µL Standard + 482 µL 1,800 720 288 115.2 46.08 18.43 7.37 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 80-fold 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 600-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 12 ml 1x Assay Diluent B to prepare a 600-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 C Rat CINC-3 concentration (pg/mL) 1 10 100 1000 10000 O D =4 50 n m 0.01 0.1 1 10 Assay Diluent B Rat CINC-3 concentration (pg/mL) 1 10 100 1000 O D =4 50 n m 0.01 0.1 1 10
Sensitivity: The minimum detectable dose of CINC-3 is typically less than 2 pg/mL.
Recovery: Recovery was determined by spiking various levels of Rat CINC-3 into serum, plasma and cell culture media. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Serum 88.29 76-101 Plasma 91.87 72-111 Cell culture media 83.48 75-89
Linearity: Sample Type Serum Plasma Cell Culture Media 1:2 Average % of Expected 93.91 91.73 104.7 Range ( %) 86-102 83-107 95-113 1:4 Average % of Expected 75.26 74.70 79.28 Range ( %) 67-83 68-83 73-86
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 (ABIN1980083) Chemokine (C-X-C Motif) Ligand 2 (CXCL2) ELISA Kit
Did you look for something else?