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Cyclin D1 Recombinant Rabbit Monoclonal Antibody (SP4)

This Antibody was verified by Knockdown to ensure that the antibody binds to the antigen stated. View details.

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Catalog # MA1-39546

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Cyclin D1 Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemical (Paraffin) analysis of Cyclin D1 using anti-Clyclin D1 antibody (Product # MA5-16356) in human mantle cell lymphoma.

Cyclin D1 Antibody in Western Blot (WB) — Western blot analysis was performed on modified whole cell extracts (1% SDS) of HeLa (Lane 1), U-87 MG (Lane 2), COLO 205 (Lane 3), A-431 (Lane 4) and NIH/3T3 (Lane 5). The blot was probed with Anti-Cyclin D1 antibody (Product # MA5-16356, 1:500 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 36 kDa band corresponding to 14-3-3 sigma was observed across the cell lines tested.

Cyclin D1 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of Cyclin D1 was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Cyclin D1 Rabbit Monoclonal Antibody (Product # MA1-39546) at 1:250 dilution in 0.1% BSA and incubated overnight at 4 degree and then labeled with Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Cyclin D1 Antibody — Antibody specificity was demonstrated by CRISPR-Cas9 mediated knockout of target protein. A reduced signal was observed for target protein in HeLa Cas9 cell line transduced with Cyclin D1 Lentiviral sgRNA compared to control cell line using Cyclin D1 Recombinant Rabbit Monoclonal Antibody (SP4) (Product # MA5-16356). {KO}

Cyclin D1 Antibody in Immunohistochemistry (IHC) — Negative correlation between ecto-enzymes and CCND1 expression in CRC. ( A ) IHC staining of human normal colon (n = 40) and colon adenocarcinoma (n = 64) specimens in a tissue microarray for CCND1 and CD163 (a macrophage marker). ( B ) Relative intensities of CCND1 staining in the lamina propria of a normal and cancerous colon. ** P < .01 (unpaired t test). ( C ) Correlation analysis of NT5E and CCND1 expression in the TCGA colorectal adenocarcinoma dataset. *** P < .001 (Student t test).

Product Details

TESTED APPLICATIONS	DILUTION
Flow Cytometry (Flow)	1:30
Immunohistochemistry (Paraffin) (IHC (P))	1:100
Western Blot (WB)	1:25-1:250
Immunocytochemistry (ICC/IF)	1:50-1:250

PUBLISHED APPLICATIONS
Immunohistochemistry (IHC)	See 229 publications below
Western Blot (WB)	See 39 publications below
Immunocytochemistry (ICC/IF)	See 8 publications below
Immunohistochemistry (Paraffin) (IHC (P))	See 19 publications below
Miscellaneous PubMed (Misc)	See 2 publications below
Immunohistochemistry (Frozen) (IHC (F))	See 2 publications below

Species reactivity	Human, Mouse, Rat
Published species	Baboon, Human, Mouse, Rat
Host / Isotype	Rabbit / IgG
Class	Recombinant Monoclonal
Type	Antibody
Clone	SP4
Immunogen	Synthetic peptide from C-terminus of human cyclin D1.
Conjugate	Unconjugated
Form	Liquid
Concentration	0.045 mg/mL
Purification	Protein A
Storage buffer	PBS, pH 7.2, with 1% BSA
Contains	0.1% sodium azide
Storage conditions	-20° C, Avoid Freeze/Thaw Cycles
RRID	AB_11000217

Product Specific Information

This antibody is predicted to react with mouse and rat based on sequence homology.

Heat-mediated antigen retrieval is recommended prior to staining, using a 10mM citrate buffer, pH 6.0, for 10 minutes followed by cooling at room temperature for 20 min. Following antigen retrieval, incubate samples with primary antibody for 30 min at room temperature. A suggested positive control is breast carcinoma or mantle cell lymphoma.

Background/Target Information

Cyclin D1 (PRAD-1, bcl-1) is one of the key cell cycle regulators, and functions in association with cdk4 and/or cdk6 by phosphorylating the Rb protein. Cyclin D1 is a putative proto-oncogene overexpressed in a wide variety of human neoplasms including mantle cell lymphomas (MCL). In addition, cyclin D1 positively regulates protein phosphorylation, mammary gland epithelial cell proliferation, and fat cell differentiation. In humans, the CCND1 gene encoding cyclin D1 is present on chromosome 11. Cyclin D1 is a cytoplasmic and nuclear protein that is synthesized during G1 phase and assembles with either cyclin-dependent kinase 4 (CDK4) or CDK6 in response to growth factor stimulation. D-type cyclin-CDK complexes act to inactivate the growth-suppressive function of the Rb protein through its phosphorylation, and titrate CDK inhibitors such as p21Cip1 and p27Kip1. Without growth factor-mediated stimulation, Cyclin D1 is unstable, and undergoes ubiquitin-mediated degradation, which is triggered by its phosphorylation. Cyclin D1 destabilization participates in G1/S phase arrest. The Cyclin D1 protein 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. Cyclin D1 forms a complex with and functions as a regulatory subunit of CDK4 or CDK6, whose activity is required for cell cycle G1/S transition. The Cyclin D1 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. Cyclin D1 has been successfully employed and is a promising tool for further studies in both cell cycle biology and cancer associated abnormalities.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

References (279)

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Bioinformatics

Protein Aliases: B-cell CLL/lymphoma 1; B-cell lymphoma 1 protein; BCL-1; BCL-1 oncogene; CCND1 protein; cDNA; FLJ93625; G1/S-specific cyclin-D1; mRNA; PRAD-1; PRAD1 oncogene

Gene Aliases: AI327039; bcl-1; BCL1; CCND1; cD1; CycD1; Cyl-1; D11S287E; PRAD1; U21B31

UniProt ID: (Human) P24385, (Rat) P39948, (Mouse) P25322

Entrez Gene ID: (Human) 595, (Rat) 58919, (Mouse) 12443

Molecular Function: Blue Arrow Icon kinase activator

Function(s)

G1/S transition of mitotic cell cycle negative regulation of transcription from RNA polymerase II promoter re-entry into mitotic cell cycle positive regulation of protein phosphorylation transcription, DNA-templated protein phosphorylation cellular response to DNA damage stimulus lactation response to iron ion response to X-ray response to organonitrogen compound positive regulation of G2/M transition of mitotic cell cycle negative regulation of Wnt signaling pathway negative regulation of epithelial cell differentiation endoplasmic reticulum unfolded protein response mitotic G1 DNA damage checkpoint response to magnesium ion response to estradiol response to vitamin E Leydig cell differentiation mammary gland epithelial cell proliferation positive regulation of mammary gland epithelial cell proliferation response to drug response to estrogen response to leptin fat cell differentiation response to ethanol positive regulation of cyclin-dependent protein serine/threonine kinase activity positive regulation of cell cycle cell division response to corticosterone response to calcium ion canonical Wnt signaling pathway mammary gland alveolus development response to UV-A negative regulation of cell cycle arrest liver regeneration liver development positive regulation of cell proliferation response to organic substance response to organic cyclic compound Wnt signaling pathway organ regeneration regulation of protein kinase activity response to steroid hormone response to glucocorticoid regulation of cell cycle cellular response to organic substance regulation of G1/S transition of mitotic cell cycle regulation of transcription, DNA-templated cell cycle

Process(es)

transcription corepressor activity
protein kinase activity
protein binding
transcription factor binding
cyclin-dependent protein serine/threonine kinase regulator activity
enzyme binding
protein kinase binding
protein complex binding
histone deacetylase binding
proline-rich region binding
kinase activity

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Cyclin D1 Recombinant Rabbit Monoclonal Antibody (SP4)

Product Details

Product Specific Information

Background/Target Information

References (279)

Have you cited this antibody in a publication? so we can reference it in this datasheet.

Bioinformatics

Suggested Secondary Antibodies

Documents

Material safety data sheets (MSDS)

Performance Guarantee

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Cyclin D1 Recombinant Rabbit Monoclonal Antibody (SP4)

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Product Details

Product Specific Information

Background/Target Information

References (279)

Bioinformatics

Suggested Secondary Antibodies

Documents

Material safety data sheets (MSDS)

Performance Guarantee

We’re here to help

Your items have has been added!