WB result of Cyclin D1 Rabbit mAb
Primary antibody: Cyclin D1 Rabbit mAb at 1/1000 dilution
Lane 1: A431 whole cell lysate 20 µg
Lane 2: MCF7 whole cell lysate 20 µg
Lane 3: LNCaP whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 33 kDa
Observed MW: 35 kDa
S-RMab® Cyclin D1 Recombinant Rabbit mAb (SDT-R423)
S-RMab® Cyclin D1 Recombinant Rabbit mAb (SDT-R423)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Synonyms | G1/S-specific cyclin-D1; B-cell lymphoma 1 protein (BCL-1); BCL-1 oncogene; PRAD1 oncogene; CCND1; BCL1; PRAD1 |
| Location | Nucleus, Cytoplasm |
| Accession | P24385 |
| Clone Number | SDT-R423 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, IHC-P, ICC, IP |
| Reactivity | Hu, Ms, Rt |
| Purification | Protein A |
| Concentration | 0.5 mg/ml |
| Conjugation | Unconjugated |
| Physical Appearance | Liquid |
| Storage Buffer | PBS, 40% Glycerol, 0.05% BSA, 0.03% Proclin 300 |
| Stability & Storage | 12 months from date of receipt / reconstitution, -20 °C as supplied |
Dilution
| application | dilution | species |
| WB | 1:1000 | |
| IP | 1:50 | |
| IHC-P | 1:500 | |
| ICC | 1:500 |
Background
Cyclin D1 is a protein that in humans is encoded by the CCND1 gene. 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 CDKs (cyclin-dependent kinase). 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 tumors and may contribute to tumorigenesis. Immunohistochemical staining of cyclin D1 antibodies is used to diagnose mantle cell lymphoma. Besides, cyclin D1 has been found to be overexpressed in breast carcinoma. Its potential use as a biomarker was suggested.
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Western Blot
WB result of Cyclin D1 Rabbit mAb
Primary antibody: Cyclin D1 Rabbit mAb at 1/1000 dilution
Lane 1: Neuro-2a whole cell lysate 20 µg
Lane 2: NIH/3T3 whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 33 kDa
Observed MW: 35 kDa
WB result of Cyclin D1 Rabbit mAb
Primary antibody: Cyclin D1 Rabbit mAb at 1/1000 dilution
Lane 1: C6 whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 33 kDa
Observed MW: 35 kDa
IP
Cyclin D1 Rabbit mAb at 1/50 dilution (1 µg) immunoprecipitating Cyclin D1 in 0.4 mg MCF7 whole cell lysate.
Western blot was performed on the immunoprecipitate using Cyclin D1 Rabbit mAb at 1/1000 dilution.
Secondary antibody (HRP) for IP was used at 1/1000 dilution.
Lane 1: MCF7 whole cell lysate 20 µg (Input)
Lane 2: Cyclin D1 Rabbit mAb IP in MCF7 whole cell lysate
Lane 3: Rabbit monoclonal IgG IP in MCF7 whole cell lysate
Predicted MW: 33 kDa
Observed MW: 33 kDa
Immunohistochemistry
IHC shows positive staining in paraffin-embedded human tonsil. Anti-Cyclin D1 antibody was used at 1/500 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human breast cancer. Anti-Cyclin D1 antibody was used at 1/500 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human colon cancer. Anti-Cyclin D1 antibody was used at 1/500 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human mantle cell lymphoma. Anti-Cyclin D1 antibody was used at 1/500 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
Immunocytochemistry
ICC shows positive staining in MCF-7 cells. Anti- Cyclin D1 antibody was used at 1/500 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 100% ice-cold methanol and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (Red).
