Recombinant
RabMAb

Anti-Cyclin D1 antibody [EPR2241] - BSA and Azide free (ab156448)

Overview

  • Product name
    Anti-Cyclin D1 antibody [EPR2241] - BSA and Azide free
    See all Cyclin D1 primary antibodies
  • Description
    Rabbit monoclonal [EPR2241] to Cyclin D1 - BSA and Azide free
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC/IF, IHC-P, IP, WBmore details
    Unsuitable for: Flow Cyt
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human Cyclin D1 (C terminal). The exact sequence is proprietary.

  • Positive control
    • Human mantle cell lymphoma tissue; MCF7 cell lysate.
  • General notes

    The formulation and the concentration of this product is compatible for metal-conjugation for mass cytometry (CyTOF®).

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab156448 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
ICC/IF Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

IP Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Predicted molecular weight: 34 kDa.
  • Application notes
    Is unsuitable for Flow Cyt.
  • Target

    • 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
    • Database links
    • 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
      • ccnd1 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
      see all

    Images

    • Immunohistochemistry was performed on human head and neck squamous cell carcinoma tissue using a rabbit monoclonal antibody against the CCND1 protein ab134175 on 3-µm slides using 224 paraffin sections via the standard SP method.

      Panel C: An example of low Cyclin D1 expression.

      Panel G: An example of high Cyclin D1 expression.

      For full image please see paper.

       

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134175).

    • Immunocytochemical analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells, labeling Cyclin D1 with ab134175 at a dilution of 1/200. Cells were paraformaldehyde fixed and permeabilized with 0.5% Triton X-100 in PBS. Incubation with the primary antibody was for 1 hour at 22°C. Cells were counterstained with DAPI following immunostaining.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134175).

    • IHC image of ab134175 staining Cyclin D1 in rat esophagus formalin fixed paraffin embedded tissue sections, performed on a Leica Bond. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab134175, 5μg/ml working concentration, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with hematoxylin and mounted with DPX. No primary antibody was used in the secondary only control (shown on the inset).
      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134175).

    • IHC image of ab134175 staining Cyclin D1 in human esophagus formalin fixed paraffin embedded tissue sections*, performed on a Leica Bond. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab134175, 5μg/ml working concentration, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with hematoxylin and mounted with DPX. No primary antibody was used in the secondary only control (shown on the inset).
      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
      *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134175).

    • ab134175 (purified) at 1/30 immunoprecipitating cyclin D1 in A431 (Human epidermoid carcinoma cell line) cells. For western blotting, an HRP-conjugated goat anti-rabbit IgG, was used as the secondary antibody (1/1000).

      Blocking/Dilution buffer and concentration: 5% NFDM/TBST.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134175).

    • Immunofluorescent staining of Ramos (Human Burkitt's lymphoma cell line) cells (fixed in 4% PFA, permeabilized with 0.1% Triton X 100) using purified ab134175 at a dilution of 1/50. An Alexa Fluor® 488 goat anti-rabbit antibody was used as the secondary at a dilution of 1/500 and the cells were counterstained with DAPI.

      The negative control is shown in the bottom right hand panel - for the negative control, Alex Fluor® 594 goat anti-mouse was used at a dilution of 1/500.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134175).

    • Immunohistochemical staining of paraffin embedded human endometrial adenocarcinoma with purified ab134175 at a dilution of 1/100. An HRP goat anti-rabbit (ab97051) was used as the secondary antibody at a dilution of 1/500 and the sample was counterstained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0.

      PBS was used instead of the primary antibody as the negative control (inset).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134175).

    • Unpurified ab134175 staining Cyclin D1 in MCF7 (Human breast adenocarcinoma cell line) cells treated with KN-93 (ab120980).

      The cells were fixed with 100% methanol (5min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab134175 at 10μg/ml and ab7291 at 1µg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with an Goat anti-Rabbit Alexa 488 secondary (ab150081) at 2 μg/ml (shown in green) and Goat anti-Mouse Alexa 594 secondary (ab150120) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labeled in blue with DAPI.

      Negative controls: 1– Rabbit primary and anti-mouse secondary antibody; 2 – Mouse primary antibody and anti-rabbit secondary antibody. Controls 1 and 2 indicate that there is no unspecific reaction between primary and secondary antibodies used.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134175).

    • Immunohistochemical analysis of paraffin-embedded human mantle cell lymphoma tissue, labeling Cyclin D1 with unpurified ab134175 at 1/100 dilution.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134175).

    • Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134175).

    References

    ab156448 has not yet been referenced specifically in any publications.

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