Overview

  • Product nameAnti-Rad21 antibodySee all Rad21 primary antibodies ...
  • Description
    Rabbit polyclonal to Rad21
  • Tested applicationsWB, ICC/IF, IHC-P more details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat, Chicken, Cow
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 350 - 450 of Human Rad21.

    (Peptide available as ab42521.)

  • Positive control
    • This antibody gave a positive signal in the following whole cell lysates: HeLa (Human epithelial carcinoma cell line), HEK293 (Human embryonic kidney cell line), A431 (Human epithelial carcinoma cell line), Jurkat (Human T cell lymphoblast-like cell line), Brain (Mouse) Tissue, Kidney (Mouse) Tissue, Heart (Mouse) Tissue, Thymus (Mouse) Tissue.

Properties

Applications

Our Abpromise guarantee covers the use of ab42522 in the following tested applications.

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

Application Notes
WB Use a concentration of 1 µg/ml. Detects a band of approximately 100 kDa (predicted molecular weight: 75 kDa).Can be blocked with Human Rad21 peptide (ab42521).
ICC/IF Use a concentration of 1 µg/ml.
IHC-P Use a concentration of 1 µg/ml.

Target

  • FunctionCleavable component of the cohesin complex, involved in chromosome cohesion during cell cycle, in DNA repair, and in apoptosis. The cohesin complex is required for the cohesion of sister chromatids after DNA replication. The cohesin complex apparently forms a large proteinaceous ring within which sister chromatids can be trapped. At metaphase-anaphase transition, this protein is cleaved by separase/ESPL1 and dissociates from chromatin, allowing sister chromatids to segregate. The cohesin complex may also play a role in spindle pole assembly during mitosis. Also plays a role in apoptosis, via its cleavage by caspase-3/CASP3 or caspase-7/CASP7 during early steps of apoptosis: the C-terminal 64 kDa cleavage product may act as a nuclear signal to initiate cytoplasmic events involved in the apoptotic pathway.
  • Sequence similaritiesBelongs to the rad21 family.
  • DomainThe C-terminal part associates with the head of SMC1A, while the N-terminal part binds to the head of SMC3.
  • Post-translational
    modifications
    Cleaved by separase/ESPL1 at the onset of anaphase. Cleaved by caspase-3 and caspase-7 at the beginning of apoptosis. The cleavage by ESPL1 and caspase-3 take place at different sites.
    Phosphorylated; becomes hyperphosphorylated in M phase of cell cycle. The large dissociation of cohesin from chromosome arms during prophase may be partly due to its phosphorylation by PLK.
  • Cellular localizationNucleus. Chromosome. Chromosome > centromere. Associates with chromatin. Before prophase it is scattered along chromosome arms. During prophase, most of cohesin complexes dissociate from chromatin probably because of phosphorylation by PLK, except at centromeres, where cohesin complexes remain. At anaphase, it is cleaved by separase/ESPL1, leading to the dissociation of the complex from chromosomes, allowing chromosome separation. Once cleaved by caspase-3, the C-terminal 64 kDa cleavage product translocates to the cytoplasm, where it may trigger apoptosis.
  • Target information above from: UniProt accession O60216 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links
  • Alternative names
    • CDLS4 antibody
    • Double strand break repair protein rad21 homolog antibody
    • Double-strand-break repair protein rad21 homolog antibody
    • FLJ25655 antibody
    • FLJ40596 antibody
    • hHR21 antibody
    • HR21 antibody
    • HRAD21 antibody
    • KIAA0078 antibody
    • KIAA0078 antibody
    • MCD1 antibody
    • Nuclear matrix protein 1 antibody
    • NXP 1 antibody
    • NXP-1 antibody
    • NXP1 antibody
    • Protein involved in DNA double-strand break repair antibody
    • Rad 21 antibody
    • RAD21 antibody
    • RAD21 homolog (S. pombe) antibody
    • RAD21 homolog antibody
    • RAD21_HUMAN antibody
    • Scc1 antibody
    • SCC1 homolog antibody
    see all

Anti-Rad21 antibody images

  • All lanes : Anti-Rad21 antibody (ab42522) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 4 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size : 75 kDa
    Observed band size : 100 kDa (why is the actual band size different from the predicted?)
    Although the predicted band size is 75kDa based on Swiss-prot data, a band of 120kDa has been previously observed.  Mol Cell Biol. 2002 Dec;22(23):8267-77 PMID: 12417729. The band seen at 100kDa is also consistent with the banding pattern observed  for other commercially available antibodies to Rad21.
  • All lanes : Anti-Rad21 antibody (ab42522) at 1 µg/ml

    Lane 1 : Brain (Mouse) Tissue Lysate
    Lane 2 : Kidney (Mouse) Tissue Lysate
    Lane 3 : Heart (Mouse) Tissue Lysate
    Lane 4 : Thymus (Mouse) Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 75 kDa
    Observed band size : 90,100 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 30 kDa (possible non-specific binding),55 kDa (possible non-specific binding).

    Exposure time : 1 minute

    The predicted molecular weight of Rad21 is 72 kDa (SwissProt), however we expect to observe a banding pattern at 100 kDa. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.

  • ICC/IF image of ab42522 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab42522, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
  • IHC image of ab42522 staining Rad21 in Human normal breast formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. 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 ab42522, 1µg/ml, 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 haematoxylin and mounted with DPX.

    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.

References for Anti-Rad21 antibody (ab42522)

ab42522 has not yet been referenced specifically in any publications.

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