Technical datasheet
Description
Monoclonal antibody MNA.1 (formerly known as 5D3-F7) recognizes human natural and recombinant monocyte chemotactic protein-1 (MCP-1). Monocyte chemotactic protein-1 (MCP-1) is a 11 kDa protein belonging to the CC subgroup of the chemokine superfamily, which stimulate the migration of monocytic cells. In contrast, the CXC chemokines predominantly activate polymorphonuclear leukocytes. The coordinated synthesis and release of MCP-1 plays a central role in both acute and chronic inflammatory processes by controlling the influx of phagocytic cells. Furthermore, their state of activation is in concert with primary inflammatory cytokines, such as IL-1, TNF-a, and IL-6. A selective accumulation of MCP-1 in the cerebrospinal fluid (CSF) of AIDS patients with cytomegalovirus encephalitis, but not with other opportunistic infections or primary lymphomas of the central nervous system , has been described. Furthermore, the chemotactic activity of MCP-1 on monocytic cells has been suggested to play a role in psoriasis, rheumatoid arthritis and atherosclerosis. No cross-reactivity of mAb MNA.1 with other cytokines has been detected.
Cross Reactivity
| Cross reactant |
Reactivity |
| Human MCP-2 |
No |
| Human MCP-3 |
No |
| Porcine MCP-1 |
Yes |
Immunogen
Recombinant human MCP-1
Formulation
1 ml (100 µg/ml) 0.2 µm filtered antibody solution in PBS, containing 0.1% bovine serum albumin.
Species
Mouse IgG1
Application
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F1,2
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FC6
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FS1
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IA1,4,8
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IF
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IP3
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P5,7,9
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W1
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Yes
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No
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N.D
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N.D.= Not Determined; F = Frozen sections; FC = Flow Cytometry; FS = Functional Studies; IA = Immuno Assays; IF = Immuno Fluorescence; IP = Immuno Precipitation; P = Paraffin sections; W = Western blot
Application notes
FC: Antibody MNA.1stains intracellular MCP-1.. For intracellular staining HUVEC cells were permeabilized with buffer containing 0.1% saponin.. The cells were fixed in 4% paraformaldehyde before staining. As negative control a corresponding isotype control antibody was used (Ref.5).
W: A reduced sample treatment and 15% SDS-Page was used. The band sizes are ~14 and 11 kDa (Ref.1).
IHC-P: Tissue sections were fixed in formalin and pretreated with hydrogen peroxide to quench endogenous peroxidases. Antigen retrieval was performed by pressure cooking for 3 minutes in PBS. As negative control the MNA.1 antibody was omitted (Ref.7).
IHC-F: Tissue sections were air dried and pretreated with 3% hydrogen peroxide to quench endogenous peroxidases. As negative control the MNA.1 antibody was omitted (Ref.2).
FS: Antibody MNA.1 inhibits migration of monocytes by neutralizing MCP-1(Ref.1).
Use
For immunohistochemistry, flow cytometry and Western blotting, dilutions to be used depend on detection system applied. It is recommended that users test the reagent and determine their own optimal dilutions. The typical starting working dilution is 1:50. For functional studies dilutions have to be optimized in user’s experimental setting.
Aliases
C-C motif chemokine 2, CCL2, small-inducible cytokine A2, MCAF, HC11
Storage and stability
Product should be stored at 4°C. Under recommended storage conditions, product is stable for at least one year. The exact expiry date is indicated on the label.
References
1. Peri, G et al; A new monoclonal antibody (5D3-F7) which recognizes human monocyte-chemotactic protein-1 but not related chemokines. Development of a sandwich ELISA and in situ detection of producing cells. J Immunol Meth 1994, 174:249
2. Wysocki, S et al; Monocyte chemoattractant protein-1 gene expression in injured pig artery coincides with early appearance of infiItrating monocyte / macrophages. J Cell Biochem 1996, 62: 303
3. Hirsch, A et al; Human cytomegalovirus inhibits transcription of the CC chemokine MCP-1 gene. J Virol 1999, 73: 404
4. Vicenzi, E et al; Divergent regulation of HIV-1 replication in PBMC of infected individuals by CC chemokines: suppression by RANTES, MIP-1, and MCP-3, and enhancement by MCP-1. J Leukoc Biol 2000, 68: 405
5. Monti, P et al; The CC Chemokine MCP-1/CCL2 in pancreatic cancer progression: regulation of expression and potential mechanisms of antimalignant activity. Cancer Res 2003, 63: 7451
6. Goebeler, M et al; Multiple signaling pathways regulate NF-kB–dependent transcription of the monocyte chemoattractant protein-1 gene in primary endothelial cells. Blood 2001, 97:46
7. Bailey, C et al; Chemokine expression is associated with the accumulation of tumour associated macrophages (TAMs) and progression in human colorectal cancer. Clin Exp Metastasis 2007, 24: 121
8. Nagarajan, S et al; Dietary soy protein isolate ameliorates atherosclerotic lesions in apolipoprotein E-deficient mice potentially by inhibiting monocyte chemoattractant protein-1 expression. J Nutr 2008, 138: 332
9. Celie, J et al; Tubulointerstitial heparan sulfate proteoglycan changes in human renal diseases correlate with leukocyte influx and proteinuria. Am J Physiol Renal Physiol 2008, 294: F253
Precautions
For research use only. Not for use in or on humans or animals or for diagnostics. It is the responsibility of the user to comply with all local/state and federal rules in the use of this product. Hycult Biotech is not responsible for any patent infringements that might result from the use or derivation of this product.
Also available
References
1. Peri, G et al; A new monoclonal antibody (5D3-F7) which recognizes human monocyte-chemotactic protein-1 but not related chemokines. Development of a sandwich ELISA and in situ detection of producing cells. J Immunol Meth 1994, 174:249
2. Wysocki, S et al; Monocyte chemoattractant protein-1 gene expression in injured pig artery coincides with early appearance of infiItrating monocyte / macrophages. J Cell Biochem 1996, 62: 303
3. Hirsch, A et al; Human cytomegalovirus inhibits transcription of the CC chemokine MCP-1 gene. J Virol 1999, 73: 404
4. Vicenzi, E et al; Divergent regulation of HIV-1 replication in PBMC of infected individuals by CC chemokines: suppression by RANTES, MIP-1, and MCP-3, and enhancement by MCP-1. J Leukoc Biol 2000, 68: 405
5. Monti, P et al; The CC Chemokine MCP-1/CCL2 in pancreatic cancer progression: regulation of expression and potential mechanisms of antimalignant activity. Cancer Res 2003, 63: 7451
6. Goebeler, M et al; Multiple signaling pathways regulate NF-kB–dependent transcription of the monocyte chemoattractant protein-1 gene in primary endothelial cells. Blood 2001, 97:46
7. Bailey, C et al; Chemokine expression is associated with the accumulation of tumour associated macrophages (TAMs) and progression in human colorectal cancer. Clin Exp Metastasis 2007, 24: 121
8. Nagarajan, S et al; Dietary soy protein isolate ameliorates atherosclerotic lesions in apolipoprotein E-deficient mice potentially by inhibiting monocyte chemoattractant protein-1 expression. J Nutr 2008, 138: 332
9. Celie, J et al; Tubulointerstitial heparan sulfate proteoglycan changes in human renal diseases correlate with leukocyte influx and proteinuria. Am J Physiol Renal Physiol 2008, 294: F253