Overview

  • Product name

    Human Cyclin D1 ELISA Kit
    See all Cyclin D1 kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Overall 5 5%
    Inter-assay
    Sample n Mean SD CV%
    Overall 3 7%
  • Sample type

    Cell culture extracts, Tissue Extracts
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    33 pg/ml
  • Range

    0.187 ng/ml - 12 ng/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture extracts 122 117% - 124%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
  • Product overview

    Cyclin D1 in vitro SimpleStep ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Cyclin D1 protein in human cell and tissue extract samples.


    The SimpleStep ELISA employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.

  • Notes

    Activity of cyclin dependent kinases CDK4 and CDK6 is regulated by the abundance of their cyclin D partners, by phosphorylation, and by association with CDK inhibitors including Kip proteins. Cyclin D-CDK4 complexes are major integrators of various mitogenenic and antimitogenic signals. The inactive ternary complex of cyclin D1/CDK4 and p27 Kip1 requires extracellular mitogenic stimuli for the dissociation and degradation of p27 concomitant with a rise in cyclin D1 levels to allow G1/S progression. Active cyclin D1-CDK4 complex phosphorylates and inhibits members of the retinoblastoma (RB) protein family including RB1. The phosphorylation of RB1 allows dissociation of the transcription factor E2F from the RB/E2F complex and the subsequent transcription of E2F target genes which are responsible for the progression through the G1 phase. Upon withdrawal of growth factors, levels of cyclin D1 protein are reduced via downregulation of protein expression and phosphorylation-dependent degradation.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Wash Buffer PT (ab206977) 1 x 20ml
    50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    ab221824 - Antibody Diluent 4BI 1 x 6ml
    10X Human Cyclin D1 Capture Antibody 1 x 600µl
    10X Human Cyclin D1 Detector Antibody 1 x 600µl
    Human Cyclin D1 Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS 1 x 12ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

  • Function

    Essential for the control of the cell cycle at the G1/S (start) transition.
  • Involvement in disease

    Note=A chromosomal aberration involving CCND1 may be a cause of B-lymphocytic malignancy, particularly mantle-cell lymphoma (MCL). Translocation t(11;14)(q13;q32) with immunoglobulin gene regions. Activation of CCND1 may be oncogenic by directly altering progression through the cell cycle.
    Note=A chromosomal aberration involving CCND1 may be a cause of parathyroid adenomas. Translocation t(11;11)(q13;p15) with the parathyroid hormone (PTH) enhancer.
    Defects in CCND1 are a cause of multiple myeloma (MM) [MIM:254500]. MM is a malignant tumor of plasma cells usually arising in the bone marrow and characterized by diffuse involvement of the skeletal system, hyperglobulinemia, Bence-Jones proteinuria and anemia. Complications of multiple myeloma are bone pain, hypercalcemia, renal failure and spinal cord compression. The aberrant antibodies that are produced lead to impaired humoral immunity and patients have a high prevalence of infection. Amyloidosis may develop in some patients. Multiple myeloma is part of a spectrum of diseases ranging from monoclonal gammopathy of unknown significance (MGUS) to plasma cell leukemia. Note=A chromosomal aberration involving CCND1 is found in multiple myeloma. Translocation t(11;14)(q13;q32) with the IgH locus.
  • Sequence similarities

    Belongs to the cyclin family. Cyclin D subfamily.
  • Post-translational
    modifications

    Phosphorylation at Thr-286 by MAP kinases is required for ubiquitination and degradation following DNA damage. It probably plays an essential role for recognition by the FBXO31 component of SCF (SKP1-cullin-F-box) protein ligase complex.
    Ubiquitinated, primarily as 'Lys-48'-linked polyubiquitination. Ubiquitinated by a SCF (SKP1-CUL1-F-box protein) ubiquitin-protein ligase complex containing FBXO4 and CRYAB (By similarity). Following DNA damage it is ubiquitinated by some SCF (SKP1-cullin-F-box) protein ligase complex containing FBXO31. Ubiquitination leads to its degradation and G1 arrest. Deubiquitinated by USP2; leading to stabilize it.
  • Cellular localization

    Nucleus.
  • Information by UniProt
  • 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
    • 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
    see all
  • Database links

Associated products

Applications

Our Abpromise guarantee covers the use of ab214571 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

  • SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

     

  • Background-subtracted data values (mean +/- SD) are graphed.

  • MCF-7 cells were grown in media containing 10% FBS (untreated), serum starved for last 24 hours (serum starved), or serum starved for last 24 hours and then treated with 10% FBS for 6 hours (serum treated). The concentrations of Cyclin D1 were measured in duplicate and interpolated from the Cyclin D1 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Cyclin D1 concentration was determined to be 8.3 ng/mL in untreated MCF-7 cell extract, 3.9 ng/mL in serum starved MCF-7 cell extract, and 11.5 ng/mL in serum treated MCF-7 cell extract.

  • Interpolated concentrations of native Cyclin D1 in human MDA-MB-231 cell extract based on a 100 µg/mL extract load, SHSY-5Y cell extract based on a 500 µg/mL extract load, and HepG2 cell extract based on a 250 µg/mL extract load. The concentrations of Cyclin D1 were measured in duplicate and interpolated from the Cyclin D1 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Cyclin D1 concentration was determined to be 0.24 ng/mL in MDA-MB-231 cell extract, 2.89 ng/mL in SHSY-5Y cell extract, and 11.7 ng/mL in HepG2 cell extract.

  • MCF-7 cells were grown in media containing 10% FBS (untreated), serum starved for last 24 hours (serum starved), or serum starved for last 24 hours and then treated with 10% FBS for 6 hours (serum treated). The concentrations of Cyclin D1 were measured in three 2-fold serial dilutions starting at 250 μg/mL in duplicates, interpolated from the Cyclin D1 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted in ng of Cyclin D1 per mg of total extracted protein (mean +/- SD, n=3). The mean Cyclin D1 concentration was determined to be 35.8 ng/mg in untreated MCF-7 cell extract, 15.7 ng/mg in serum starved MCF-7 cell extract, and 48.2 ng/mg in serum treated MCF-7 cell extract.

  • The concentrations of Cyclin D1 were measured in three 2-fold serial dilutions starting at 500 µg/mL for NIH/3T3 and C2C12 cell extracts and 250 µg/mL for H9C2 cell extracts in duplicates, interpolated from the human Cyclin D1 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted in ng of cyclin D1 per mg of total extracted protein (mean +/- SD, n=2-3). The mean Cyclin D1 concentration was determined to be 1.7 ng/mg in NIH/3T3 cell extract, 23.1 ng/mg in C2C12 cell extract, and 43.1 ng/mg in H9C2 cell extract.

Protocols

References

This product has been referenced in:

  • Gaballah HH  et al. Expression of long non-coding RNA CCHE1 in colorectal carcinoma: correlations with clinicopathological features and ERK/COX-2 pathway. Mol Biol Rep N/A:N/A (2018). Read more (PubMed: 30484108) »
See 1 Publication for this product

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