Product Information

Contents: Biotin anti-mouse/human/rat MCP-1
Catalog Number: 13-7096
Sizes: 50 ug, 500 ug
Formulation: Phosphate buffer pH 7.2, 150 mM NaCl, 0.09% NaN3
Storage Conditions: Store at 4°C. Avoid repeated freeze/thaw cycles.
Clone: 2H5
Isotype: Armenian Hamster IgG

Mouse thiogylcolate induced PEC were stimulated with LPS in the presence of Brefeldin A for 24 hours. (Left) The cells were surface stained with FITC anti-mouse F4/80 antigen (BM8) and intracellularly stained with PE rat IgG2a isotype control. (Right) The cells were surface stained with FITC anti-mouse F4/80 antigen and intracellularly stained with PE anti-mouse CCL2 (2H5).

Available Formats of This Product
Cat. No.	Format	Excite (nm)	Emit (nm)	Reported Applications
11-7096	Fluorescein isothiocyanate (FITC) anti-mouse/human/rat CCL2 (MCP-1, MCP1, MCAF)	488	518	IC Flow
12-7096	Phycoerythrin (PE) anti-mouse/human/rat CCL2 (MCP-1, MCP1, MCAF)	488	575	IC Flow
13-7096	Biotin anti-mouse/human/rat CCL2 (MCP-1, MCP1, MCAF)	N/A	N/A	ELISA det
14-7096	Affinity Purified anti-mouse/human/rat CCL2 (MCP-1, MCP1, MCAF)	N/A	N/A	ELISA det IC Flow IH/F WB
16-7096	Functional Grade* Purified anti-mouse/human/rat CCL2 (MCP-1, MCP1, MCAF)	N/A	N/A	NU
*Functional Grade™ (FG™) Purified: Azide-free, sterile-filtered, and endotoxin < 0.001 ng/µg (unless otherwise noted). *Functional Grade™ (FG™) Biotin: Azide-free, sterile-filtered, and endotoxin < 0.05 ng/µg. Purified: Contains azide, not sterile-filtered, and not endotoxin tested.

Questions? Please consult our answers to frequently asked questions at http://www.ebioscience.com/faq.

Description

The 2H5 monoclonal antibody reacts with mouse, rat, and human monocyte chemoattractant protein-1 (MCP-1), also known as CCL2 and MCAF.

Applications Reported

For research use only, not for diagnostic or therapeutic use. The 2H5 antibody has been reported for use in ELISA, intracellular staining for flow cytometric analysis, and cytokine neutralization. (Please use Functional Grade purified 2H5 antibody, cat. 16-7096, in functional assays.)

Applications Tested

Mouse MCP-1 ELISA: The biotinylated 2H5 antibody has been tested as the detection antibody in a sandwich ELISA for analysis of mouse MCP-1 in combination with the affinity purified 4E2/MCP (14-7091) antibody for capture and recombinant mouse MCP-1 as the standard. A suitable range of concentrations of this antibody for ELISA detection is 0.5-2.0 µg/ml. A standard curve consisting of doubling dilutions of the recombinant standard over the range of 2000 pg/ml - 15 pg/ml should be included in each ELISA plate.
Human MCP-1 ELISA: The biotinylated 2H5 antibody has been tested as the detection antibody in a sandwich ELISA for analysis of human MCP-1 in combination with the affinity purified 5D3-F7 (14-7099) antibody for capture and recombinant human MCP-1 as the standard. A suitable range of concentrations of this antibody for ELISA detection is 0.5-2.0 µg/ml. A standard curve consisting of doubling dilutions of the recombinant standard over the range of 2000 pg/ml - 15 pg/ml should be included in each ELISA plate.
The Functional Grade purified 2H5 antibody has been tested for neutralization of MCP-1 bioactivity.
The fluorochrome-conjugated 2H5 antibody has been tested for intracellular staining and flow cytometric analysis of mouse, rat, and human cells. The fluorochrome-conjugated 2H5 antibody is offered in 2 formats:
- µg size: has been tested by intracellular staining and flow cytometric analysis and can be used at less than or equal to 0.5 μg/million cells. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.
- test size: has been pre-titrated and tested by intracellular staining and flow cytometric analysis and can be used at 20 μl per million cells in a 100 μl total staining volume.

References

Luo, Y., et al. 1994. J Immunol. 153: 3708. [ Antibody generation, ELISA, immunoblotting, in vitro neutralization]

Morrison, B., et al. 2003. J. Clin. Invest. 112: 1862. [In vivo neutralization]

Taal MW, Chertow GM, et al. 2001. Mechanisms underlying renoprotection during renin-angiotensin system blockade. Am J Physiol Renal Physiol. 280(2):F343-55. (2H5, IHC frozen in rat, PubMed)

Luo Y, Fischer FR, et al. 2000. Macrophage inflammatory protein-2 and KC induce chemokine production by mouse astrocytes. J Immunol. 165(7):4015-23. (2H5, IHC frozen, PubMed)

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