WB
Western blot analysis of Cyclin D1 on different lysates with Rabbit anti-Cyclin D1 antibody at 1/5000 dilution and competitor's antibody at 1/5000 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. Exposure time: 20 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 at 1/5000 dilution and competitor's antibody at 1/5000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50000 dilution was used for 1 hour at room temperature.WB
Western blot analysis of Cyclin D1 on different lysates with Rabbit anti-Cyclin D1 antibody at 1/5000 dilution. Lane 1: MCF7-si NT cell lysate, Lane 2: MCF7-si Cyclin D1 cell lysate, Lysates/proteins at 10 µg/Lane. 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 at 1/5000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50000 dilution was used for 1 hour at room temperature.ICC/IF
Immunocytochemistry analysis of Neuro-2a cells labeling Cyclin D1 with Rabbit anti-Cyclin D1 antibody at 1/2000 dilution and competitor's antibody at 1/1600 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 at 1/2000 dilution and competitor's antibody at 1/1600 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1000 dilution.IHC
Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-Cyclin D1 antibody at 1/1000 dilution. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins. The section was incubated with Rabbit anti-Cyclin D1 antibody for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.IHC
Immunohistochemical analysis of paraffin-embedded mouse stomach tissue with Rabbit anti-Cyclin D1 antibody at 1/1000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) 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 at 1/1000 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.IHC
Immunohistochemical analysis of paraffin-embedded rat esophagus tissue with Rabbit anti-Cyclin D1 antibody at 1/1000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) 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 at 1/1000 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.IP
Cyclin D1 was immunoprecipitated from 0.2 mg MCF7 cell lysate with Rabbit anti-Cyclin D1 antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using Rabbit anti-Cyclin D1 antibody at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5000 dilution was used for 1 hour at room temperature. Lane 1: MCF7 cell lysate (input), Lane 2: Rabbit anti-Cyclin D1 antibody IP in MCF7 cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-Cyclin D1 antibody in MCF7 cell lysate, Blocking/Dilution buffer: 5% NFDM/TBST, Exposure time: 5 seconds.IF-P
Application: IF-Tissue, Species: Human, Site: colon cancer, Sample: Paraffin-embedded section, Antibody concentration: 1/500.| Product Name | Cyclin D1 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Synthetic peptide within C-terminal human Cyclin D1. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | ICC/IFIF-PIHCIPWB |
| WB:1:5000 ICC/IF:1:2000 IHC:1:200-1:1000 FC:1:5000 IF-P: 1:500 IP:Use at an assay dependent concentration. |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Affinity purified |
| Gene Symbol | CCND1 |
|---|---|
| Gene Synonyms | BCL1 PRAD1 U21B31 D11S287E |
| Gene Full Name | cyclin D1 |
| Gene Summary | The protein encoded by this gene belongs to the highly conserved cyclin family, whose members are characterized by a dramatic periodicity in protein abundance throughout the cell cycle. Cyclins function as regulators of CDK kinases. Different cyclins exhibit distinct expression and degradation patterns which contribute to the temporal coordination of each mitotic event. This cyclin forms a complex with and functions as a regulatory subunit of CDK4 or CDK6, whose activity is required for cell cycle G1/S transition. This protein has been shown to interact with tumor suppressor protein Rb and the expression of this gene is regulated positively by Rb. Mutations, amplification and overexpression of this gene, which alters cell cycle progression, are observed frequently in a variety of human cancers. [provided by RefSeq, Dec 2019] |
| Alternative Names | AI327039 antibody B cell CLL/lymphoma 1 antibody B cell leukemia 1 antibody B cell lymphoma 1 protein antibody B-cell lymphoma 1 protein antibody BCL 1 antibody BCL-1 antibody BCL-1 oncogene antibody BCL1 antibody BCL1 oncogene antibody AI327039 antibody B cell CLL/lymphoma 1 antibody B cell leukemia 1 antibody B cell lymphoma 1 protein antibody B-cell lymphoma 1 protein antibody BCL 1 antibody BCL-1 antibody BCL-1 oncogene antibody BCL1 antibody BCL1 oncogene antibody ccnd1 antibody cyclind1 antibody CCND1/FSTL3 fusion gene included antibody CCND1/IGHG1 fusion gene included antibody CCND1/IGLC1 fusion gene included antibody CCND1/PTH fusion gene included antibody CCND1_HUMAN antibody cD1 antibody Cyl 1 antibody D11S287E antibody G1/S specific cyclin D1 antibody G1/S-specific cyclin-D1 antibody Parathyroid adenomatosis 1 antibody PRAD1 antibody PRAD1 oncogene antibody U21B31 antibody |
| Molecular Weight(MW) | 34kDa(Observed band size:35kDa) |
| Cellular Localization | Cytoplasm, Nucleus, Membrane, Mitochondrion |
WB
Western blot analysis of Cyclin D1 on different lysates with Rabbit anti-Cyclin D1 antibody at 1/5000 dilution and competitor's antibody at 1/5000 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. Exposure time: 20 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 at 1/5000 dilution and competitor's antibody at 1/5000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50000 dilution was used for 1 hour at room temperature.
WB
Western blot analysis of Cyclin D1 on different lysates with Rabbit anti-Cyclin D1 antibody at 1/5000 dilution. Lane 1: MCF7-si NT cell lysate, Lane 2: MCF7-si Cyclin D1 cell lysate, Lysates/proteins at 10 µg/Lane. 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 at 1/5000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50000 dilution was used for 1 hour at room temperature.
ICC/IF
Immunocytochemistry analysis of Neuro-2a cells labeling Cyclin D1 with Rabbit anti-Cyclin D1 antibody at 1/2000 dilution and competitor's antibody at 1/1600 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 at 1/2000 dilution and competitor's antibody at 1/1600 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1000 dilution.
IHC
Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-Cyclin D1 antibody at 1/1000 dilution. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins. The section was incubated with Rabbit anti-Cyclin D1 antibody for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
IHC
Immunohistochemical analysis of paraffin-embedded mouse stomach tissue with Rabbit anti-Cyclin D1 antibody at 1/1000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) 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 at 1/1000 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.
IHC
Immunohistochemical analysis of paraffin-embedded rat esophagus tissue with Rabbit anti-Cyclin D1 antibody at 1/1000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) 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 at 1/1000 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.
IP
Cyclin D1 was immunoprecipitated from 0.2 mg MCF7 cell lysate with Rabbit anti-Cyclin D1 antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using Rabbit anti-Cyclin D1 antibody at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5000 dilution was used for 1 hour at room temperature. Lane 1: MCF7 cell lysate (input), Lane 2: Rabbit anti-Cyclin D1 antibody IP in MCF7 cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-Cyclin D1 antibody in MCF7 cell lysate, Blocking/Dilution buffer: 5% NFDM/TBST, Exposure time: 5 seconds.
IF-P
Application: IF-Tissue, Species: Human, Site: colon cancer, Sample: Paraffin-embedded section, Antibody concentration: 1/500.| Application Notes | WB:1:5000 ICC/IF:1:2000 IHC:1:200-1:1000 FC:1:5000 IF-P: 1:500 IP:Use at an assay dependent concentration. |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | 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 | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
