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Human MIP-1 alpha/CCL3 ELISA Kit, Fluorescent is a single-wash 90-min SimpleStep ELISA® for the quantitative measurement of Human MIP-1 alpha/CCL3 , Fluorescent in Heparin Plasma, Citrate plasma, Cell culture supernatant, Serum, EDTA Plasma samples.

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Images

Sandwich ELISA - Human MIP-1 alpha/CCL3 ELISA Kit, Fluorescent (AB229400), expandable thumbnail
  • Sandwich ELISA - Human MIP-1 alpha/CCL3 ELISA Kit, Fluorescent (AB229400), expandable thumbnail
  • Sandwich ELISA - Human MIP-1 alpha/CCL3 ELISA Kit, Fluorescent (AB229400), expandable thumbnail
  • Sandwich ELISA - Human MIP-1 alpha/CCL3 ELISA Kit, Fluorescent (AB229400), expandable thumbnail

Key facts

Detection method

Fluorescent

Sample types

Heparin Plasma, Citrate plasma, Cell culture supernatant, Serum, EDTA Plasma

Assay type

Sandwich (quantitative)

Reactive species

Human

Range

0.2 - 200 pg/mL

Assay time

1h 30m

Sensitivity

= 0.2 pg/mL

Reactivity data

Application

sELISA

Reactivity

Reacts

Dilution info
-
Notes

-

Associated Products

Select an associated product type

2 products for Alternative Product

Target data

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).

Alternative names

What's included?

1 x 96 Tests
Components
100X Stoplight Red Substrate
1 x 120 µL
10X Human MIP-1 alpha/CCL3 Capture Antibody
1 x 600 µL
10X Human MIP-1 alpha/CCL3 Detector Antibody
1 x 600 µL
10X Wash Buffer PT (ab206977)
1 x 20 mL
500X Hydrogen Peroxide (H2O2, 3%)
1 x 50 µL
Antibody Diluent CPI - HAMA Blocker (ab193969)
1 x 6 mL
Human MIP-1 alpha/CCL3 Lyophilized Recombinant Protein
2 x 1 Vial
Plate Seals
1 x 1 Unit
Sample Diluent NS (ab193972)
1 x 50 mL
SimpleStep Pre-Coated Black 96-Well Microplate
1 x 1 Unit
Stoplight Red Substrate Buffer
1 x 12 mL

Recommended products

Human MIP-1 alpha/CCL3 ELISA Kit, Fluorescent is a single-wash 90-min SimpleStep ELISA® for the quantitative measurement of Human MIP-1 alpha/CCL3 , Fluorescent in Heparin Plasma, Citrate plasma, Cell culture supernatant, Serum, EDTA Plasma samples.

Key facts

Detection method

Fluorescent

Sample types

Heparin Plasma, Citrate plasma, Cell culture supernatant, Serum, EDTA Plasma

Assay type

Sandwich (quantitative)

Reactive species

Human

Range

0.2 - 200 pg/mL

Assay time

1h 30m

Assay Platform

Pre-coated microplate (12 x 8 well strips)

Sensitivity

= 0.2 pg/mL

Precision

Intra assay

Sample

Supernatant

n
5
mean
-
SD
-
C.V.

1.9

Inter assay

Sample

Supernatant

n
3
mean
-
SD
-
C.V.

7.2

Recovery

Sample specific recovery

Sample type

Cell culture supernatant

Average %

= 101

Range

100 - 102 %

Sample type

Serum

Average %

= 99

Range

96 - 101 %

Sample type

EDTA Plasma

Average %

= 100

Range

92 - 105 %

Sample type

Heparin Plasma

Average %

= 97

Range

94 - 100 %

Sample type

Citrate plasma

Average %

= 95

Range

91 - 101 %

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

+4°C

Storage information

+4°C

Notes

MIP-1 alpha/CCL3 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.

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).

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

MIP-1 alpha also known as CCL3 is a chemokine that plays a critical role in the immune system. With a molecular weight of around 8 kDa it is primarily expressed in activated T-cells macrophages and fibroblasts. MIP-1 alpha acts as a signaling protein that binds to receptors on target cells directing the movement of immune cells. Researchers commonly refer to it using various alternative names such as MIP-1a and CCL3-alpha rat and it is frequently measured using assays like MIP-1 alpha ELISA for experimental analyses.

Biological function summary

Proteins including MIP-1 alpha participate in the recruitment and activation of diverse immune cells aiding the inflammatory response. Through interaction with receptors like CCR1 and CCR5 MIP-1 alpha mobilizes cells like natural killer cells and monocytes. This chemokine does not generally form part of a larger protein complex but interacts directly with membrane receptors to exert its function in immune cell chemotaxis.

Pathways

MIP-1 alpha operates in the context of both the inflammatory response and the chemokine signaling pathways. Its involvement in these pathways helps mediate the trafficking of leukocytes. Notably MIP-1 alpha interacts with proteins such as CCR1 and CCR5 to transmit signals that instigate immune responses. These proteins are integral to its role in modulating immunity providing targets for research into controlling inflammatory processes.

Associated diseases and disorders

MIP-1 alpha closely relates to conditions such as rheumatoid arthritis and HIV infection. In rheumatoid arthritis its overexpression can lead to excessive inflammation and joint destruction. Meanwhile in HIV infection MIP-1 alpha interacts with CCR5 a co-receptor necessary for the virus entry into cells. This dual relationship elucidates its potential as a therapeutic target in managing both inflammatory diseases and viral entry mechanisms.

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4 product images

  • Sandwich ELISA - Human MIP-1 alpha/CCL3 ELISA Kit, Fluorescent (ab229400), expandable thumbnail

    Sandwich ELISA - Human MIP-1 alpha/CCL3 ELISA Kit, Fluorescent (ab229400)

    Example of human MIP1a standard curve in Sample Diluent NS.

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

  • Sandwich ELISA - Human MIP-1 alpha/CCL3 ELISA Kit, Fluorescent (ab229400), expandable thumbnail

    Sandwich ELISA - Human MIP-1 alpha/CCL3 ELISA Kit, Fluorescent (ab229400)

    Interpolated concentrations of spiked MIP1a in human serum and plasma samples.

    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).

  • Sandwich ELISA - Human MIP-1 alpha/CCL3 ELISA Kit, Fluorescent (ab229400), expandable thumbnail

    Sandwich ELISA - Human MIP-1 alpha/CCL3 ELISA Kit, Fluorescent (ab229400)

    Serum from ten individual healthy human female donors was measured in duplicate.

    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.

  • Sandwich ELISA - Human MIP-1 alpha/CCL3 ELISA Kit, Fluorescent (ab229400), expandable thumbnail

    Sandwich ELISA - Human MIP-1 alpha/CCL3 ELISA Kit, Fluorescent (ab229400)

    Interpolated concentrations of native MIP1a in human peripheral blood monocyte (PBMC) cell culture supernatant samples.

    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.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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