CCL3 / MIP-1 alpha Recombinant Rabbit Monoclonal Antibody [PSH11-28]
cat.: HA723310
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse |
| Applications: | WB, IF-Cell, IHC-P |
| Clonality: | Monoclonal |
| Clone number: | PSH11-28 |
| Form: | Liquid |
| Storage condition: | Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term. |
| Storage buffer: | 1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 10 kDa |
| Isotype: | IgG |
| Immunogen: | Recombinant protein within human CCL3 aa 1-92. |
| Positive control: | THP-1 treated with 100nM TPA overnight then add 100ng/mL LPS for 7 hours then add 1μg/mL BFA for 3 hours cell lysate, RAW264.7 treated with 100nM TPA overnight then add 100ng/mL LPS for 7 hours then add 1μg/mL BFA for 3 hours cell lysate, NK-92 cell lysate, RAW264.7 cells treated with 100nM TPA overnight then add 100ng/mL LPS for 7 hours, human hodgkin lymphoma tissue, human spleen tissue. |
| Subcellular location: | Secreted. |
| Recommended Dilutions:
WB IF-Cell IHC-P |
1:2,000 1:2,000 1:200 |
| Uniprot #: | SwissProt: P10147 Human | P10855 Mouse |
| Alternative names: | C C motif chemokine 3 CCL 3 CCL3 CCL3_HUMAN Chemokine (C C motif) ligand 3 Chemokine C C motif ligand 3 Chemokine ligand 3 G0/G1 switch regulatory protein 19 1 G0/G1 switch regulatory protein 19-1 G0S19 1 G0S19 1 protein Heparin binding chemotaxis protein L2G25B LD78 alpha LD78-alpha(4-69) LD78alpha Macrophage inflammatory protein 1 alpha Macrophage inflammatory protein 1-alpha macrophage inflammatory protein 1a MIP 1 alpha MIP 1A MIP-1-alpha MIP-1-alpha(4-69) MIP1 alpha MIP1A PAT 464.1 SCYA 3 SCYA3 SIS alpha SIS beta SIS-beta Small inducible cytokine A3 small inducible cytokine A3 (homologous to mouse Mip-1a) Small-inducible cytokine A3 Tonsillar lymphocyte LD78 alpha protein |
Images
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Fig1:
Western blot analysis of CCL3 / MIP-1 alpha on different lysates with Rabbit anti-CCL3 / MIP-1 alpha antibody (HA723310) at 1/2,000 dilution. Lane 1: THP-1 cell lysate Lane 2: THP-1 treated with 100nM TPA overnight then add 100ng/mL LPS for 7 hours then add 1μg/mL BFA for 3 hours cell lysate Lane 3: RAW264.7 cell lysate Lane 4: RAW264.7 treated with 100nM TPA overnight then add 100ng/mL LPS for 7 hours then add 1μg/mL BFA for 3 hours cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 10 kDa Observed band size: 10 kDa Exposure time: 25 seconds; ECL: K1802; 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 (HA723310) at 1/2,000 dilution was used in 5% NFDM/TBST 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:
Western blot analysis of CCL3 / MIP-1 alpha on different lysates with Rabbit anti-CCL3 / MIP-1 alpha antibody (HA723310) at 1/2,000 dilution. Lane 1: NK-92 cell lysate (20 µg/Lane) Lane 2: HeLa cell lysate (negative) (20 µg/Lane) Predicted band size: 10 kDa Observed band size: 10 kDa Exposure time: 25 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 (HA723310) at 1/2,000 dilution was used in 5% NFDM/TBST 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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Fig3:
Immunocytochemistry analysis of RAW264.7 cells untreated / treated with 100nM TPA overnight then add 100ng/mL LPS for 7 hours labeling CCL3 / MIP-1 alpha with Rabbit anti-CCL3 / MIP-1 alpha antibody (HA723310) at 1/2,000 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-CCL3 / MIP-1 alpha antibody (HA723310) at 1/2,000 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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Fig4:
Immunohistochemical analysis of paraffin-embedded human hodgkin lymphoma tissue with Rabbit anti-CCL3 / MIP-1 alpha antibody (HA723310) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723310) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. |
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Fig5:
Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-CCL3 / MIP-1 alpha antibody (HA723310) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723310) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.