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Human CXCR2(CXC-Chemokine Receptor 2) ELISA Kit (HUES03382)

SKU:
HUES03382
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
P25025
Sensitivity:
0.47ng/mL
Range:
0.78-50ng/mL
ELISA Type:
Sandwich
Synonyms:
CD182,CXCR2,IL8R2,CMKAR2,CDw128b,IL8RB
Reactivity:
Human
Sample Type:
Serum, plasma and other biological fluids
Research Area:
Cell Biology
€599
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Description

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Assay type:Sandwich
Format:96T
Assay time:4.5h
Reactivity:Human
Detection Method:Colormetric
Detection Range:0.78-50 ng/mL
Sensitivity:0.47 ng/mL
Sample Volume Required Per Well:100µL
Sample Type:Serum, plasma and other biological fluids
Specificity:This kit recognizes Human CXCR2 in samples. No significant cross-reactivity or interference between Human CXCR2 and analogues was observed.

This ELISA kit uses Sandwich-ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human CXCR2. Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human CXCR2 and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human CXCR2, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by adding Stop Solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human CXCR2. The concentration of Human CXCR2 in samples can be calculated by comparing the OD of the samples to the standard curve.

UniProt Protein Function:IL8RB: Receptor for interleukin-8 which is a powerful neutrophil chemotactic factor. Binding of IL-8 to the receptor causes activation of neutrophils. This response is mediated via a G-protein that activates a phosphatidylinositol-calcium second messenger system. Binds to IL-8 with high affinity. Also binds with high affinity to CXCL3, GRO/MGSA and NAP-2. Belongs to the G-protein coupled receptor 1 family.
UniProt Protein Details:

Protein type:GPCR, family 1; Receptor, cytokine; Membrane protein, multi-pass; Membrane protein, integral; Receptor, GPCR

Chromosomal Location of Human Ortholog: 2q35

Cellular Component: cell surface; integral to plasma membrane; intracellular; mast cell granule; membrane; plasma membrane

Molecular Function:C-X-C chemokine receptor activity; interleukin-8 binding; interleukin-8 receptor activity; protein binding; signal transducer activity

Biological Process: cell surface receptor linked signal transduction; cellular defense response; chemotaxis; dendritic cell chemotaxis; G-protein signaling, coupled to IP3 second messenger (phospholipase C activating); inflammatory response; neutrophil activation; neutrophil chemotaxis; positive regulation of cell proliferation; receptor internalization; signal transduction

NCBI Summary:The protein encoded by this gene is a member of the G-protein-coupled receptor family. This protein is a receptor for interleukin 8 (IL8). It binds to IL8 with high affinity, and transduces the signal through a G-protein activated second messenger system. This receptor also binds to chemokine (C-X-C motif) ligand 1 (CXCL1/MGSA), a protein with melanoma growth stimulating activity, and has been shown to be a major component required for serum-dependent melanoma cell growth. This receptor mediates neutrophil migration to sites of inflammation. The angiogenic effects of IL8 in intestinal microvascular endothelial cells are found to be mediated by this receptor. Knockout studies in mice suggested that this receptor controls the positioning of oligodendrocyte precursors in developing spinal cord by arresting their migration. This gene, IL8RA, a gene encoding another high affinity IL8 receptor, as well as IL8RBP, a pseudogene of IL8RB, form a gene cluster in a region mapped to chromosome 2q33-q36. Alternatively spliced variants, encoding the same protein, have been identified. [provided by RefSeq, Nov 2009]
UniProt Code:P25025
NCBI GenInfo Identifier:1352454
NCBI Gene ID:3579
NCBI Accession:P25025. 2
UniProt Secondary Accession:P25025,Q8IUZ1, Q9P2T6, Q9P2T7,
UniProt Related Accession:P25025,AAB24742,AAA15417
Molecular Weight:40,759 Da
NCBI Full Name:C-X-C chemokine receptor type 2
NCBI Synonym Full Names:C-X-C motif chemokine receptor 2
NCBI Official Symbol:CXCR2
NCBI Official Synonym Symbols:CD182; IL8R2; IL8RA; IL8RB; CMKAR2; CDw128b
NCBI Protein Information:C-X-C chemokine receptor type 2
UniProt Protein Name:C-X-C chemokine receptor type 2
UniProt Synonym Protein Names:CDw128b; GRO/MGSA receptor; High affinity interleukin-8 receptor B; IL-8R B; IL-8 receptor type 2; CD_antigen: CD182
Protein Family:C-X-C chemokine receptor
UniProt Gene Name:CXCR2
UniProt Entry Name:CXCR2_HUMAN

As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e. g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.

Concentration
(ng/mL)		O.DAverageCorrected
502.359
2.395		2.3772.314
251.637
1.639		1.6381.575
12.50.874
0.848		0.8610.798
6.250.394
0.408		0.4010.338
3.130.261
0.253		0.2570.194
1.570.16
0.158		0.1590.096
0.780.108
0.116		0.1120.049
00.054
0.072		0.063--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human CXCR2 were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human CXCR2 were tested on 3 different plates, 20 replicates in each plate.

Intra-assay PrecisionInter-assay Precision
Sample123123
n202020202020
Mean (ng/mL)2.765.7224.542.575.8623.93
Standard deviation0.150.270.870.130.260.87
C V (%)5.434.723.555.064.443.64

Recovery

The recovery of Human CXCR2 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample TypeRange (%)Average Recovery (%)
Serum (n=5)93-10599
EDTA plasma (n=5)93-10999
Cell culture media (n=5)89-10094

Linearity

Samples were spiked with high concentrations of Human CXCR2 and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Serum (n=5)EDTA plasma (n=5)Cell culture media (n=5)
1:2Range (%)87-9898-11094-107
Average (%)92103101
1:4Range (%)93-10484-9688-103
Average (%)999094
1:8Range (%)87-10085-9788-102
Average (%)929094
1:16Range (%)94-10581-9384-98
Average (%)998790

An unopened kit can be stored at 4°C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.

Item Specifications Storage
Micro ELISA Plate(Dismountable) 8 wells ×12 strips -20°C, 6 months
Reference Standard 2 vials
Concentrated Biotinylated Detection Ab (100×) 1 vial, 120 µL
Concentrated HRP Conjugate (100×) 1 vial, 120 µL -20°C(shading light), 6 months
Reference Standard & Sample Diluent 1 vial, 20 mL 4°C, 6 months
Biotinylated Detection Ab Diluent 1 vial, 14 mL
HRP Conjugate Diluent 1 vial, 14 mL
Concentrated Wash Buffer (25×) 1 vial, 30 mL
Substrate Reagent 1 vial, 10 mL 4°C(shading light)
Stop Solution 1 vial, 10 mL 4°C
Plate Sealer 5 pieces
Product Description 1 copy
Certificate of Analysis 1 copy
  1. Set standard, test sample and control (zero) wells on the pre-coated plate and record theirpositions. It is recommended to measure each standard and sample in duplicate. Note: addall solutions to the bottom of the plate wells while avoiding contact with the well walls. Ensuresolutions do not foam when adding to the wells.
  2. Aliquot 100µl of standard solutions into the standard wells.
  3. Add 100µl of Sample / Standard dilution buffer into the control (zero) well.
  4. Add 100µl of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids) into test sample wells.
  5. Cover the plate with the sealer provided in the kit and incubate for 90 min at 37°C.
  6. Aspirate the liquid from each well, do not wash. Immediately add 100µL of BiotinylatedDetection Ab working solution to each well. Cover the plate with a plate seal and gently mix. Incubate for 1 hour at 37°C.
  7. Aspirate or decant the solution from the plate and add 350µL of wash buffer to each welland incubate for 1-2 minutes at room temperature. Aspirate the solution from each well andclap the plate on absorbent filter paper to dry. Repeat this process 3 times. Note: a microplatewasher can be used in this step and other wash steps.
  8. Add 100µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37°C.
  9. Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
  10. Add 90µL of Substrate Reagent to each well. Cover with a new plate seal and incubate forapproximately 15 min at 37°C. Protect the plate from light. Note: the reaction time can beshortened or extended according to the actual color change, but not by more than 30min.
  11. Add 50 µL of Stop Solution to each well. Note: Adding the stop solution should be done inthe same order as the substrate solution.
  12. Determine the optical density (OD value) of each well immediately with a microplate readerset at 450 nm.

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