Mouse MCP-1(CCL2) Recombinant

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Mouse MCP-1(CCL2) Recombinant

$70.00 – $2,700.00

SKU: RKP10148 Category: Recombinant Mouse Cytokines Tags: CCL2, Je, mcp-1, mcp1, P10148, SCYA2

Description

Methods (9)


accession	P10148

Source	Optimized DNA sequence encoding Mouse MCP-1(CCL2) mature chain was expressed in Escherichia Coli.
Molecular weight	Mouse MCP-1, generated by the proteolytic removal of the signal peptide and propeptide and has a calculated molecular mass of approximately9 kDa. RecombinantMCP-1 is a monomeric protein consisting of 125 amino acid residue subunits and migrates as an approximately 14 kDa protein under non-reducing conditions and reducing conditions in SDS-PAGE.
Purity	>98%, as determined by SDS-PAGE and HPLC
Biological Activity	Determined by its ability to chemoattract Balb/C mouse spleen MNCs using a concentration range of 1.0-20.0 ng/ml.
Endotoxin	Endotoxin content was assayed using a LAL gel clot method. Endotoxin level was found to be less than 0.1 ng/µg(1EU/µg).
Presentation	MCP-1was lyophilized from PBS pH 7.4.
Reconstitution	A quick spin of the vial followed by reconstitution in distilled water to a concentration not less than 0.1 mg/mL. This solution can then be diluted into other buffers.
Storage	This cytokine can be stored in working aliquots at 2° - 8° C for one month, or at -20° C for six months, with a carrier protein without detectable loss of activity. Avoid repeated freeze/thaw cycles.
Usage	This cytokine product is for research purposes only.It may not be used for therapeutics or diagnostic purposes.

Interactor	E9M5R0
Biological Process	Chemotaxis
Biological Process	Inflammatory-response
Molecular function	Cytokine

Methods


Epithelial cytokine expression and MSC migration. B/ Supernatants from non-infected (white bars), 7.13- (light grey bars) or 26695- (dark grey bars) infected cells were assessed for TNFα and CCL2 expression by ELISA.
Cell Migration The upper chamber wells of CIM-Plates 16 were precoated with fibronectin, added to wells at 10 µg/ml (30 µl) and removed after 30 min. The lower chamber wells were filled with 160 µl culture medium, allowing a clearly-defined meniscus to form. SDF1, MCP1 and TNFα were added to the lower chambers at a final concentration of 100 ng/ml. CIM-Plates were equilibrated for at least 30 min in a culture hood at room temperature. Trypsin-EDTA-dispersed MPCs (100 µl in complete medium) were then plated in the fibronectin-precoated upper chamber wells at 306×10³ cells/cm² (600000 cells/ml) in 100 µl complete medium, which we determined as the optimal density to ensure confluent monolayers. The CIM-Plates were inserted into the cradle pockets of the RTCA DP Analyzer, and the background impedance of each well was measured after 1 h. The RTCA DP Analyzer, placed in a standard cell-culture incubator, monitored the migrated cell number… The upper chamber wells of CIM-Plates 16 were precoated with fibronectin, added to wells at 10 µg/ml (30 µl) and removed after 30 min. The lower chamber wells were filled with 160 µl culture medium, allowing a clearly-defined meniscus to form. SDF1, MCP1 and TNFα were added to the lower chambers at a final concentration of 100 ng/ml. CIM-Plates were equilibrated for at least 30 min in a culture hood at room temperature. Trypsin-EDTA-dispersed MPCs (100 µl in complete medium) were then plated in the fibronectin-precoated upper chamber wells at 306×10³ cells/cm² (600000 cells/ml) in 100 µl complete medium, which we determined as the optimal density to ensure confluent monolayers. The CIM-Plates were inserted into the cradle pockets of the RTCA DP Analyzer, and the background impedance of each well was measured after 1 h. The RTCA DP Analyzer, placed in a standard cell-culture incubator, monitored the migrated cell number in each lower chamber well every 15 min (quadruplicate measurements) over 36 h. Data were analyzed with the RTCA Software. Read more
Transwell Migration Assay The bottom chambers of 24-well transwell plates (8 µm pore polycarbonate membrane transwell, ) contained 600 µl of each assaying condition in complete media (RPMI +10% FBS, for THP-1 cells) or minimal media (DMEM for RAW264.7 cells). For THP-1 cells the bottom chambers of the transwells containing RPMI media were untreated or contained PKRA7 (1 µg/ml), MCP-1 (100 ng/ml), MCP-1+ PKRA7, PK2 (200 ng/ml), or PK2+ PKRA7 in triplicate per condition. For RAW264.7 cells the bottom chambers of the transwells containing DMEM media were untreated or contained PKRA7 (1 µg/ml), SDF-1α (200 ng/ml), SDF-1α+PKRA7, (200 ng/ml), or + PKRA7 in triplicate per condition. MCP-1, SDF-1α and PK2 all dissolved in water plus 0.1% BSA (water +0.1% BSA used as control). The appropriate number of cells was collected in complete (THP-1) or minimal media (RAW264.7) and 100 µl were plated onto the… The bottom chambers of 24-well transwell plates (8 µm pore polycarbonate membrane transwell, ) contained 600 µl of each assaying condition in complete media (RPMI +10% FBS, for THP-1 cells) or minimal media (DMEM for RAW264.7 cells). For THP-1 cells the bottom chambers of the transwells containing RPMI media were untreated or contained PKRA7 (1 µg/ml), MCP-1 (100 ng/ml), MCP-1+ PKRA7, PK2 (200 ng/ml), or PK2+ PKRA7 in triplicate per condition. For RAW264.7 cells the bottom chambers of the transwells containing DMEM media were untreated or contained PKRA7 (1 µg/ml), SDF-1α (200 ng/ml), SDF-1α+PKRA7, (200 ng/ml), or + PKRA7 in triplicate per condition. MCP-1, SDF-1α and PK2 all dissolved in water plus 0.1% BSA (water +0.1% BSA used as control). The appropriate number of cells was collected in complete (THP-1) or minimal media (RAW264.7) and 100 µl were plated onto the top chamber. The transwell plates containing the cells were placed in an incubator and the cells were allowed to migrate for 6 hours (THP-1) or 18 hours (RAW264.7). The cells were then fixed with 4% PFA, stained with 0.5% toluidine blue in 4% PFA, counted using a microscope and analyzed. Read more
PKRA7 decreases subcutaneous and intracranial glioblastoma xenograft tumor growth. D456MG cells were SC injected into nude mice, and control (n = 5) or PKRA7 (n = 5) treatment was commenced when tumors became visually detectable (14 days).
Altered myelopoiesis in Sgpl1chimeras. Bone marrow-derived macrophages of Sgpl1chimeras are unresponsive to S1P stimulus as assessed by Ca2+-flux measurement, while the Ca2+ response to ionomycin and MCP-1 remained unaltered (n = 3 per group).
Validation of the array expression pattern by quantitative RT-PCR. The mRNA levels of CCL2/MCP-1 , CCL5/RANTES , ICAM-1 , VCAM-1 , E-selectin , and P-selectin were determined by quantitative real-time PCR, and values were normalized to 18S rRNA as reference gene.
HIV Infection Significantly Increases Monocyte Transmigration Across the BBB in Response to CCL2. “Day 3” monocytes from 8 separate individuals were infected with HIV or remained uninfected as described in the methods, added to our BBB model, and allowed to transmigrate for 24 hours to media alone or to 200 ng/mL CCL2.
Characterization of macrophage-involvement in GVHD. CCL2 mRNA expression was evaluated using quantitative real-time RT-PCR.
Immunophenotypical and functional maturation of DCs. C) IL-6, MCP-1, IL-1β and TNFα release from CASMC, iDCs, mDCs, ccDCs as assayed by Milliplex method.