Overview

  • Product name

  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Overall 5 1.44%
    Inter-assay
    Sample n Mean SD CV%
    Overall 3 1.97%
  • Sample type

    Cell culture supernatant, Urine, Serum, Hep Plasma, EDTA Plasma, Cit plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    1.95 pg/ml
  • Range

    15.6 pg/ml - 1000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Urine 81 75% - 85%
    Serum 81 80% - 83%
    Cell culture media 87 91% - 93%
    Cit plasma 82 78% - 86%
    serum free media 95 93% - 96%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Mouse
    Does not react with: Goat, Cow, Pig
  • Product overview

    Abcam’s MIP1a (CCL3) in vitro SimpleStep ELISA™ (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of mouse MIP1a protein in serum, plasma, urine and cell culture supernatant.

    The SimpleStep ELISA™ employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.

  • Notes

    Macrophage Inflammatory Protein 1 alpha (MIP1a, also known as C-C motif chemokine 3 or CCL3) is a secreted monokine with inflammatory and chemokinetic properties. Mature MIP1a is 68 amino acids after cleavage of a 23 amino acid signal peptide. MIP1a can be induced in a variety of cell types and for immune and inflammatory response. In humans, MIP1a is a major HIV-suppressive factor produced by CD8+ T cells. The standard protein in this kit is mature mouse MIP1a.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Mouse MIP1a Capture Antibody 1 x 600µl
    10X Mouse MIP1a Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    Antibody Diluent 5BI 1 x 6ml
    Mouse MIP1a Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS (ab193972) 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

  • Function

    Monokine with inflammatory and chemokinetic properties. Binds to CCR1, CCR4 and CCR5. One of the major HIV-suppressive factors produced by CD8+ T-cells. Recombinant MIP-1-alpha induces a dose-dependent inhibition of different strains of HIV-1, HIV-2, and simian immunodeficiency virus (SIV).
  • Sequence similarities

    Belongs to the intercrine beta (chemokine CC) family.
  • Post-translational
    modifications

    N-terminal processed form LD78-alpha(4-69) is produced by proteolytic cleavage after secretion from HTLV1-transformed T-cells.
  • Cellular localization

    Secreted.
  • Information by UniProt
  • Alternative names

    • C C motif chemokine 3
    • CCL 3
    • CCL3
    • CCL3_HUMAN
    • Chemokine (C C motif) ligand 3
    • Chemokine C C motif ligand 3
    • Chemokine ligand 3
    • G0/G1 switch regulatory protein 19 1
    • G0/G1 switch regulatory protein 19-1
    • G0S19 1
    • G0S19 1 protein
    • Heparin binding chemotaxis protein
    • L2G25B
    • LD78 alpha
    • LD78-alpha(4-69)
    • LD78alpha
    • Macrophage inflammatory protein 1 alpha
    • Macrophage inflammatory protein 1-alpha
    • macrophage inflammatory protein 1a
    • MIP 1 alpha
    • MIP 1A
    • MIP-1-alpha
    • MIP-1-alpha(4-69)
    • MIP1 alpha
    • MIP1A
    • PAT 464.1
    • SCYA 3
    • SCYA3
    • SIS alpha
    • SIS beta
    • SIS-beta
    • Small inducible cytokine A3
    • small inducible cytokine A3 (homologous to mouse Mip-1a)
    • Small-inducible cytokine A3
    • Tonsillar lymphocyte LD78 alpha protein
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab200017 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

  • SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

     

  • Background-subtracted data values (mean +/- SD) are graphed.

  • RAW264.7 cells were grown in the absence (unstimulated) or presence of LPS (stimulated) for 2 days. MIP1a was measured in 2-fold diluted cell culture supernatants of unstimulated and LPS stimulated RAW264.7 and cell culture media. Measured values were interpolated from the MIP1a standard curve diluted in Sample Diluent NS and corrected for dilution factor. Mean of duplicate values +/-SD are graphed: 20 ng/mL unstimulated, 118 ng/mL stimulated, and undetectable in media.

  • The concentrations of MIP1a were measured in duplicate and interpolated from the MIP1a standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean MIP1a concentration was determined to be 118 µg/mL in RAW264.7 LPS Stimulated supernatant.

Protocols

References

This product has been referenced in:

See 1 Publication for this product

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