CCL2 / MCP-1 Rabbit Polyclonal Antibody
cat.: HA500267
| Product Type: | Rabbit polyclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Mouse |
| Applications: | WB, IF-Cell, FC |
| Clonality: | Polyclonal |
| Form: | Liquid |
| Storage condition: | Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term. |
| Storage buffer: | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 1ug/ul |
| Purification: | Immunogen affinity purified. |
| Molecular weight: | Predicted band size: 16 kDa |
| Isotype: | IgG |
| Immunogen: | Recombinant protein within rat CCL2 aa 1-148. |
| Positive control: | RAW264.7 treated with 100nM TPA overnight then 100ng/mL LPS for 7 hours add 1μg/mL BFA for last 3 hours cell lysate, RAW264.7 cells treated with 100nM TPA overnight then 100ng/mL LPS for 7 hours add 1μg/mL BFA for last 3 hours. |
| Subcellular location: | Secreted. |
| Recommended Dilutions:
WB IF-Cell FC |
1:5,000 1:500 1:1,000 |
| Uniprot #: | SwissProt: P10148 Mouse |
| Alternative names: | C-C motif chemokine 2 CCL2 Chemokine (C C motif) ligand 2 GDCF 2 GDCF-2 GDCF2 HC11 HSMCR30 JE MCAF MCP 1 MCP-1 MCP1 MGC9434 Monocyte chemoattractant protein 1 Monocyte chemotactic and activating factor Monocyte chemotactic protein 1 Monocyte secretory protein JE SCYA2 Small inducible cytokine A2 (monocyte chemotactic protein 1, homologous to mouse Sig je) Small inducible cytokine A2 Small inducible cytokine subfamily A (Cys Cys), member 2 Small-inducible cytokine A2 SMC CF SMC-CF SMCCF |
Images
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Fig1:
Western blot analysis of CCL2 / MCP-1 on different lysates with Rabbit anti-CCL2 / MCP-1 antibody (HA500267) at 1/5,000 dilution. Lane 1: RAW264.7 cell lysate Lane 2: RAW264.7 treated with 100nM TPA overnight then 100ng/mL LPS for 7 hours add 1μg/mL BFA for last 3 hours cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 16 kDa Observed band size: 16 kDa Exposure time: 10 seconds; ECL: K1801; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA500267) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig2:
Immunocytochemistry analysis of RAW264.7 cells untreated / treated with 100nM TPA overnight then 100ng/mL LPS for 7 hours add 1μg/mL BFA for last 3 hours labeling CCL2 / MCP-1 with Rabbit anti-CCL2 / MCP-1 antibody (HA500267) at 1/500 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-CCL2 / MCP-1 antibody (HA500267) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
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Fig3:
Flow cytometric analysis of RAW264.7 cells untreated (left) / treated with 100nM TPA overnight then 100ng/mL LPS for 7 hours add 1μg/mL BFA for last 3 hours (right) labeling CCL2 / MCP-1. Cells were fixed and permeabilized. Then stained with the primary antibody (HA500267, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.