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Rat MIP-1α(Macrophage Inflammatory Protein 1 Alpha) ELISA Kit

  • Cat.No.:E-EL-R0602

  • Reactivity: Rat

To Purchase E-EL-R0602

Size:
  • 96T
  • 48T
  • 24T
  • 96T*5
Price: $609
Qty:

Product Details

Properties

Assay type Sandwich-ELISA
Format 96T
Assay time 3.5h
Detection range 31.25-2000 pg/mL
Sensitivity 18.75 pg/mL
Sample type &Sample volume Serum, plasma and other biological fluids; 100μL
Specificity This kit recognizes Rat MIP-1α in samples. No significant cross-reactivity or interference between Rat MIP-1α and analogues was observed.
Reproducibility Both intra-CV and inter-CV are < 10%.
Application This ELISA kit applies to the in vitro quantitative determination of Rat MIP-1α 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 Rat MIP-1α. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Rat MIP-1α 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 Rat MIP-1α, 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 Rat MIP-1α. You can calculate the concentration of Rat MIP-1α in the samples by comparing the OD of the samples to the standard curve.

Kit components & Storage

An unopened kit can be stored at 2-8℃ for 1 month. If the kit is not supposed to be used within 1 month, store the components separately according to the following conditions once the kit is received.
ItemSpecificationsStorage
Micro ELISA Plate(Dismountable) 96T: 8 wells ×12 strips
48T: 8 wells ×6 strips
-20℃, 6 months
Reference Standard 96T: 2 vials
48T: 1 vial
Concentrated Biotinylated Detection Ab (100×) 96T: 1 vial, 120 μL
48T: 1 vial, 60 μL
Concentrated HRP Conjugate (100×) 96T: 1 vial, 120 μL
48T: 1 vial, 60 μL
-20℃(Protect from light), 6 months
Reference Standard & Sample Diluent 1 vial, 20 mL 2-8°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 2-8℃(Protect from light)
Stop Solution 1 vial, 10 mL 2-8°C
Plate Sealer 5 pieces
Manual 1 copy
Certificate of Analysis 1 copy

Technical Data

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Rat MIP-1α were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Rat MIP-1α 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) 104.60 298.50 924.40 97.00 296.80 958.20
Standard deviation 5.20 12.80 36.10 5.10 14.50 38.30
CV (%) 4.97 4.29 3.91 5.26 4.89 4.00

Recovery

The recovery of Rat MIP-1α 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=8) 88-103 94
EDTA plasma (n=8) 95-107 102
Cell culture media (n=8) 90-105 96

Target Information

Database LinksSwissProt:   P50229
SynonymsCCL3, LD78ALPHA, MIP-1-alpha, MIP1A, SCYA3, G0S19-1
Research AreaImmunology

Assay Procedures

elisa assay procedure 1

1. Add 100μL standard or sample to the wells. Incubate for 90 min at 37°C

elisa assay procedure 2

2. Discard the liquid, immediately add 100μL Biotinylated Detection Ab working solution to each well. Incubate for 60 min at 37°C

elisa assay procedure 3

3. Aspirate and wash the plate for 3 times

elisa assay procedure 4

4. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times

elisa assay procedure 5

5. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C

elisa assay procedure 6

6. Add 50μL Stop Solution

elisa assay procedure 7

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

Citations

  1. International Journal of Biological Sciences (2022) IF: 6.582
    Sublytic C5b-9 Induces CCL3/4 Production and Macrophage Accumulation in Thy-1N Rats via PKC-α/p65/IRF-8 Axis

    DOI: 10.7150/ijbs.69652

    PMID: 35637950

    Sample: renal tissue,GMCs
  2. EXPERIMENTAL AND MOLECULAR MEDICINE (2017) IF: 5.063
    TGF-β1 suppresses CCL3/4 expression through the ERK signaling pathway and inhibits intervertebral disc degeneration and inflammation-related pain in a rat model

    DOI: 10.1038/emm.2017.136

    Sample: Cell culture supernatant
  3. International Journal of Biological Sciences (2019) IF: 4.067
    Hydrogen sulfide treatment alleviated ventilator-induced lung injury through regulation of autophagy and endoplasmic reticulum stress

    DOI: 10.7150/ijbs.38315

    PMID: 31853224

    Sample: Serum,Bronchoalveolar Lavage Fluid,Tissue homogena
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