Overview

  • Product name
  • Detection method
    Fluorescent
  • Precision
    Intra-assay
    Sample n Mean SD CV%
    Supernatant 5 1.9%
    Inter-assay
    Sample n Mean SD CV%
    Supernatant 3 7.2%
  • Sample type
    Cell culture supernatant, Serum, Heparin Plasma, EDTA Plasma, Citrate Plasma
  • Assay type
    Sandwich (quantitative)
  • Sensitivity
    0.2 pg/ml
  • Range
    0.2 pg/ml - 200 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 101 100% - 102%
    Serum 99 96% - 101%
    Heparin Plasma 97 94% - 100%
    EDTA Plasma 100 92% - 105%
    Citrate Plasma 95 91% - 101%

  • Assay time
    1h 30m
  • Assay duration
    One step assay
  • Species reactivity
    Reacts with: Human
    Does not react with: Rat, Cow
  • Product overview

    MIP1a in vitro CatchPoint® SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of MIP1a protein in human serum, plasma and cell culture supernatant.


    This CatchPoint SimpleStep ELISA kit has been optimized for Molecular Devices Microplate Readers. Click here for a list of recommended Microplate Readers.
    If using a Molecular Devices’ plate reader supported by SoftMax® Pro software, a preconfigured protocol for these CatchPoint SimpleStep ELISA Kits is available with all the protocol and analysis settings at www.softmaxpro.org.


    The CatchPoint® 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. CatchPoint® HRP Development Solution containing the Stoplight Red Substrate is added. During incubation, the substrate is catalyzed by HRP generating a fluorescent product. Signal is generated proportionally to the amount of bound analyte and the intensity is measured in a fluorescence plater reader at 530/570/590 nm Excitation/Cutoff/Emission.


    16% reactivity observed in mouse.

  • Notes

    Macrophage Inflammatory Protein 1-alpha (MIP1a, also known as CCL3) is a monokine with both inflammatory and chemokine properties. MIP-1-alpha can bind to CCR1, CCR4 and CCR5. In addition, it is 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).

  • Tested applications
    Suitable for: Sandwich ELISAmore details
  • Platform
    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions
    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    100X Stoplight Red Substrate 1 x 120µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    500X Hydrogen Peroxide (H2O2, 3%) 1 x 50µl
    Antibody Diluent CPI 1 x 6ml
    10X Human MIP1a Capture Antibody 1 x 600µl
    10X Human MIP1a Detector Antibody 1 x 600µl
    Human MIP1a Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS 1 x 50ml
    SimpleStep Pre-Coated Black 96-Well Microplate 1 unit
    Stoplight Red Substrate Buffer 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
    • 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
    • MIP 1 alpha
    • MIP 1A
    • MIP-1-alpha
    • MIP-1-alpha(4-69)
    • 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 ab229400 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

  • ELISA Protocol Summary
  • The MIP1a standard curve was prepared as described in Section 10. Background-subtracted data values (mean +/- SD) are graphed.

  • The concentrations of MIP1a were measured in duplicates, interpolated from the MIP1a standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 50%, plasma (EDTA) 50%, plasma (heparin) 50%) and plasma (citrate) 25%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).

  • Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean MIP1a concentration was determined to be 5.5 pg/mL with a range of 1.3 – 12.1 pg/mL.

  • The concentrations of MIP1a were measured in duplicates, interpolated from the MIP1a standard curves and corrected for sample dilution. Undiluted samples are as follows: unstimulated 6.35% and stimulated 0.2%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean MIP1a concentration was determined to be 0.241 ng/mL in unstimulated PBMC cell culture supernatant and 40.8 ng/mL in stimulated.

Protocols

References

ab229400 has not yet been referenced specifically in any publications.

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