Anti-Cyclin D1 Monoclonal Antibody (5D4)

Product Name

Anti-Cyclin D1 Monoclonal Antibody (5D4)

View all CCND1 Antibodies

SKU/Catalog Number

M00149-2

Size

100µg

Form

Liquid. In PBS, pH 7.2, containing 50% glycerol.

Description

Boster Bio Anti-Cyclin D1 Monoclonal Antibody (5D4) catalog # M00149-2. Tested in Flow Cytometry, IP, IF, IHC, WB applications. This antibody reacts with Human, Mouse.

Storage & Handling

Store at -20°C for one year. For short term storage and frequent use, store at 4°C for up to one month. Avoid repeated freeze-thaw cycles.

Cite This Product

Anti-Cyclin D1 Monoclonal Antibody (5D4) (Boster Biological Technology, Pleasanton CA, USA, Catalog # M00149-2)

Host

Mouse

Contents

Liquid. In PBS, pH 7.2, containing 50% glycerol.

Clonality

Monoclonal

Clone Number

5D4

Isotype

IgG2a, kappa

Immunogen

Recombinant human cyclin D1.

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Reactive Species

M00149-2 is reactive to CCND1 in Human, Mouse

Applications

M00149-2 is guaranteed for Flow Cytometry, IP, IF, IHC, WB Boster Guarantee

Observed Molecular Weight

Calculated molecular weight

33.729kDa

Background of CCND1

Recognizes a non-histone protein of 36kDa, which is identified as proliferating cell nuclear antigen (PCNA). It is also known as cyclin or polymerase delta auxiliary protein. Elevated expression of PCNA/cyclin has been shown in the nucleus during late G1 phase immediately before the onset of DNA synthesis, becoming maximal during S-phase and declining during G2 and M phases. This monoclonal antibody is excellent for multiple applications.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconsitution

Restore with deionized water (or equivalent) for reconstitution volume of 100 µL

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Flow Cytometry (10µg/ml final concentration)
Immunohistochemistry (10µg/ml, frozen/paraffin)
Immunoprecipitation (1-10µg/200µg cell extract from 5x10⁶ cells)
Western Blot (1µg/ml, ECL)
Suggested dilutions/conditions may not be available for all applications.
Optimal conditions must be determined individually for each application.

Validation Images & Assay Conditions

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Immunoprecipitation analysis of cyclin D1 from ZR75-1 cells with Mouse IgG2a (1) or Cyclin D1 mAb (5D4) (2). After immunoprecipitated with the antibody, immunocomplex was resolved on SDS-PAGE and immunoBlotted with 553.

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Flow cytometry analysis of Cyclin DIsotypic control to the cells. Shaded histogram indicates the reaction of Cyclin D1 mAb (5D4) to the cells.

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Figure 1. Western blot analysis of CCND1 using anti-CCND1 antibody (M00149-2).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-CCND1 antigen affinity purified polyclonal antibody (Catalog # M00149-2) at 0.5 ug/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-Mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # SA1021) with Tanon 5200 system. A specific band was detected for CCND1.

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Figure 4. IHC analysis of CCND1 using anti-CCND1 antibody (M00149-2).
CCND1 was detected in paraffin-embedded section. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-CCND1 Antibody (M00149-2) overnight at 4°C. Biotinylated goat anti Mouse IgG antibody was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1021) with DAB as the chromogen.

Gene/Protein Information For CCND1 (Source: Uniprot.org, NCBI)

Alternative Names

B-cell lymphoma 1 protein; BCL-1 oncogene; Bcl-1; BCL1D11S287E; CCND1; cyclin D1 (PRAD1: parathyroid adenomatosis 1); Cyclin D1; Cyl-1; G1/S-specific cyclin D1; G1/S-specific cyclin-D1; PRAD1 oncogene; PRAD1; PRAD1B-cell CLL/lymphoma 1; U21B31 CCND1 BCL1, D11S287E, PRAD1, U21B31 cyclin D1 G1/S-specific cyclin-D1|B-cell CLL/lymphoma 1|B-cell lymphoma 1 protein|BCL-1 oncogene|PRAD1 oncogene

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In this infographic, you will see the following information for CCND1: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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