Cyclin D1 Recombinant Rabbit Monoclonal Antibody [PD01-64]
cat.: HA721322
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IHC-P, IF-Cell, FC |
| Clonality: | Monoclonal |
| Clone number: | PD01-64 |
| 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: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 34 kDa |
| Isotype: | IgG |
| Immunogen: | Recombinant protein within Human Cyclin D1 aa 200-295 (C terminal). |
| Positive control: | MCF7 cell lysate, A431 cell lysate, Neuro-2a cell lysate, NIH/3T3 cell lysate, C6 cell lysate, SH-SY5Y cell lysate, Neuro-2a, MCF7, human colon carcinoma tissue, human testis tissue, NIH/3T3. |
| Subcellular location: | Cytoplasm, Nucleus, Membrane, Mitochondrion |
| Recommended Dilutions:
WB IHC-P IF-Cell FC |
1:5,000 1:200 1:2,000 1:5,000 |
| Uniprot #: | SwissProt: P24385 Human | P25322 Mouse | P39948 Rat |
| Alternative names: | AI327039 B cell CLL/lymphoma 1 B cell leukemia 1 B cell lymphoma 1 protein B-cell lymphoma 1 protein BCL 1 BCL-1 BCL-1 oncogene BCL1 BCL1 oncogene ccnd1 cyclind1 CCND1/FSTL3 fusion gene CCND1/FSTL3 fusion gene, included CCND1/IGHG1 fusion gene CCND1/IGHG1 fusion gene, included CCND1/IGLC1 fusion gene CCND1/IGLC1 fusion gene, included CCND1/PTH fusion gene CCND1/PTH fusion gene, included CCND1_HUMAN cD1 Cyl 1 D11S287E G1/S specific cyclin D1 G1/S-specific cyclin-D1 Parathyroid adenomatosis 1 PRAD1 PRAD1 oncogene U21B31 |
Images
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Fig1:
Western blot analysis of Cyclin D1 on different lysates with Rabbit anti-Cyclin D1 antibody (HA721322) at 1/5,000 dilution and competitor's antibody at 1/5,000 dilution. Lane 1: MCF7 cell lysate Lane 2: K-562 cell lysate (negative) Lane 3: A431 cell lysate Lane 4: Neuro-2a cell lysate Lane 5: NIH/3T3 cell lysate Lane 6: C6 cell lysate Lane 7: SH-SY5Y cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 34 kDa Observed band size: 35 kDa Exposure time: 20 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 (HA721322) at 1/5,000 dilution and competitor's antibody at 1/5,000 dilution were 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:
Immunocytochemistry analysis of Neuro-2a cells labeling Cyclin D1 with Rabbit anti-Cyclin D1 antibody (HA721322) at 1/2,000 dilution and competitor's antibody at 1/1,600 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-Cyclin D1 antibody (HA721322) at 1/2,000 dilution and competitor's antibody at 1/1,600 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 (M1305-2, 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 MCF7 cells labeling Cyclin D1. Cells were fixed and permeabilized. Then stained with the primary antibody (HA721322, red) at 1/5,000 dilution and competitor's antibody (red) at 1/2,000 dilution, 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). |
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Fig4:
Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-Cyclin D1 antibody (HA721322) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (HA721322) 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 testis tissue with Rabbit anti-Cyclin D1 antibody (HA721322) at 1/50 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (HA721322) at 1/50 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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Fig6:
Immunocytochemistry analysis of NIH/3T3 cells labeling Cyclin D1 with Rabbit anti-Cyclin D1 antibody (HA721322) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-Cyclin D1 antibody (HA721322) at 1/50 dilution in 2% negative goat serum 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. |
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Fig7:
Western blot analysis of Cyclin D1 on different lysates with Rabbit anti-Cyclin D1 antibody (HA721322) at 1/5,000 dilution. Lane 1: MCF7-si NT cell lysate Lane 2: MCF7-si Cyclin D1 cell lysate Lysates/proteins at 10 µg/Lane. Predicted band size: 34 kDa Observed band size: 35 kDa Exposure time: 17 seconds; 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 (HA721322) at 1/5,000 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. |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.