Human MCP-1(Monocyte Chemotactic Protein 1) ELISA Kit-Laboratory Biological Research Reagents Manufacturer for Life Science - Elabscience

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Human MCP-1(Monocyte Chemotactic Protein 1) ELISA Kit

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Price:   $609
Cat Number:  E-EL-H6005
Lead Time:   Order now, ship in 3 days

Reactivity:  Human

Detection Range:  62.50~4000 pg/mL

Sensitivity:  37.50 pg/mL

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Intended use

This ELISA kit applies to the in vitro quantitative determination of Human MCP-1 concentrations in serum, plasma and other biological fluids.

Test principle

This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human MCP-1. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human MCP-1 and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human MCP-1, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of 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 MCP-1. You can calculate the concentration of Human MCP-1 in the samples by comparing the OD of the samples to the standard curve.

Assay type Sandwich-ELISA
Format 96T/48T
Assay time 3.5h
Reactivity Human
Detection Method Colormetric
Detection Range 62.50—4000 pg/mL
Sensitivity 37.50 pg/mL
Sample Volume Required Per Well 100μL
Sample Type serum, plasma and other biological fluids

Specificity

This kit recognizes Human MCP-1 in samples. No significant cross-reactivity or interference between Human MCP-1 and analogues was observed.

Typical data

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
(pg/mL)
O.D Average Corrected
4000 2.552
2.507
2.53 2.469
2000 1.634
1.645
1.64 1.579
1000 0.957
0.946
0.952 0.891
500 0.487
0.506
0.497 0.436
250 0.275
0.268
0.272 0.211
125 0.19
0.19
0.19 0.129
62.50 0.129
0.113
0.121 0.06
0 0.057
0.064
0.061 --
sandwich-Ab-ELISA-Elabscience

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human MCP-1 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human MCP-1 were tested on 3 different plates, 20 replicates in each plate.

Intra-assay Precision Inter-assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 198.91 488.33 1680.55 164.36 433.06 1730.78
Standard deviation 10.01 24.12 90.58 9.45 20.74 76.85
C V (%) 5.03 4.94 5.39 5.75 4.79 4.44

Recovery

The recovery of Human MCP-1 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample Type Range (%) Average Recovery (%)
Serum (n=5) 97-108 101
EDTA plasma (n=5) 89-99 94
Cell culture media (n=5) 87-95 92

Linearity

Samples were spiked with high concentrations of Human MCP-1 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)
1:2 Range (%) 98-110 89-102 91-104
Average (%) 103 96 98
1:4 Range (%) 87-98 90-101 94-100
Average (%) 92 94 97
1:8 Range (%) 90-100 94-103 101-111
Average (%) 94 99 105
1:16 Range (%) 89-99 84-96 89-101
Average (%) 96 92 97

Kit components & Storage

An unopened kit can be stored at 4℃ 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℃, 6 months
Reference Standard 2 vials
Concentrated Biotinylated Detection Ab (100×) 1 vial, 120 μL
Concentrated HRP Conjugate (100×) 1 vial, 120 μL -20℃(shading light), 6 months
Reference Standard & Sample Diluent 1 vial, 20 mL 4℃, 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℃(shading light)
Stop Solution 1 vial, 10 mL 4℃
Plate Sealer 5 pieces
Product Description 1 copy
Certificate of Analysis 1 copy

Note: All reagent bottle caps must be tightened to prevent evaporation and microbial pollution.
The volume of reagents in partial shipments is a little more than the volume marked on the label, please use accurate measuring equipment instead of directly pouring into the vial(s).

Other supplies required

  • Microplate reader with 450 nm wavelength filter
  • High-precision transfer pipette, EP tubes and disposable pipette tips
  • Incubator capable of maintaining 37℃
  • Deionized or distilled water
  • Absorbent paper
  • Loading slot for Wash Buffer

Assay Procedure

1.Add 100 μL standard or sample to each well. Incubate for 90 min at 37℃.

2.Remove the liquid.

3.Add 100 μL Biotinylated Detection Ab. Incubate for 1 hour at 37℃. Aspirate and wash 3 times.

4.Add 100 μL HRP Conjugate. Incubate for 30 min at 37℃. Aspirate and wash 5 times.

5.Add 90 μL of Substrate Reagent. Incubate for 15 min at 37℃.

6.Add 50 μL Stop Solution.

7.Read at 450 nm immediately. Calculation of results.

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