Overview

  • Product name
  • Description
    Goat polyclonal to Artemis
  • Host species
    Goat
  • Specificity
    This antibody detects a band of the appropriate size in a number of different cell lines. However, it does not detect a band in cell lines that do not express artemis, it has been tested in the CJ cell line (see below) and in an hTERT line.
  • Tested applications
    Suitable for: IHC-P, WBmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide:

    CPKDTYSDLKSRDK

    , corresponding to amino acids 478-490 of Human Artemis.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 0.42% Potassium phosphate, 0.87% Sodium chloride
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab3834 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
IHC-P 1/1000.
WB 1/500. Detects a band of approximately 90 kDa (predicted molecular weight: 78 kDa).

Target

  • Function
    Required for V(D)J recombination, the process by which exons encoding the antigen-binding domains of immunoglobulins and T-cell receptor proteins are assembled from individual V, (D), and J gene segments. V(D)J recombination is initiated by the lymphoid specific RAG endonuclease complex, which generates site specific DNA double strand breaks (DSBs). These DSBs present two types of DNA end structures: hairpin sealed coding ends and phosphorylated blunt signal ends. These ends are independently repaired by the non homologous end joining (NHEJ) pathway to form coding and signal joints respectively. This protein exhibits single-strand specific 5'-3' exonuclease activity in isolation and acquires endonucleolytic activity on 5' and 3' hairpins and overhangs when in a complex with PRKDC. The latter activity is required specifically for the resolution of closed hairpins prior to the formation of the coding joint. May also be required for the repair of complex DSBs induced by ionizing radiation, which require substantial end-processing prior to religation by NHEJ.
  • Tissue specificity
    Ubiquitously expressed, with highest levels in the kidney, lung, pancreas and placenta (at the mRNA level). Expression is not increased in thymus or bone marrow, sites of V(D)J recombination.
  • Involvement in disease
    Defects in DCLRE1C are a cause of severe combined immunodeficiency autosomal recessive T-cell-negative/B-cell-negative/NK-cell-positive with sensitivity to ionizing radiation (RSSCID) [MIM:602450]. SCID refers to a genetically and clinically heterogeneous group of rare congenital disorders characterized by impairment of both humoral and cell-mediated immunity, leukopenia, and low or absent antibody levels. Patients with SCID present in infancy with recurrent, persistent infections by opportunistic organisms. The common characteristic of all types of SCID is absence of T-cell-mediated cellular immunity due to a defect in T-cell development. Individuals affected by RS-SCID show defects in the DNA repair machinery necessary for coding joint formation and the completion of V(D)J recombination. A subset of cells from such patients show increased radiosensitivity.
    Defects in DCLRE1C are the cause of severe combined immunodeficiency Athabaskan type (SCIDA) [MIM:602450]. SCIDA is a variety of RS-SCID caused by a founder mutation in Athabascan-speaking native Americans, being inherited as an autosomal recessive trait with an estimated gene frequency of 2.1% in the Navajo population. Affected individuals exhibit clinical symptoms and defects in DNA repair comparable to those seen in RS-SCID.
    Defects in DCLRE1C are a cause of Omenn syndrome (OS) [MIM:603554]. OS is characterized by severe combined immunodeficiency associated with erythrodermia, hepatosplenomegaly, lymphadenopathy and alopecia. Affected individuals have elevated T-lymphocyte counts with a restricted T-cell receptor (TCR) repertoire. They also generally lack B-lymphocytes, but have normal natural killer (NK) cell function (T+ B- NK+).
  • Sequence similarities
    Belongs to the DNA repair metallo-beta-lactamase (DRMBL) family.
  • Post-translational
    modifications
    Phosphorylation on undefined residues by PRKDC may stimulate endonucleolytic activity on 5' and 3' hairpins and overhangs. PRKDC must remain present, even after phosphorylation, for efficient hairpin opening. Also phosphorylated by ATM in response to ionizing radiation (IR) and by ATR in response to ultraviolet (UV) radiation.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • A SCID antibody
    • A SCID protein antibody
    • Artemis protein antibody
    • ASCID antibody
    • DCLRE1C antibody
    • DCLRE1C DNA cross link repair 1C antibody
    • DCLRE1C protein antibody
    • DCLREC1C antibody
    • DCR1C_HUMAN antibody
    • DNA cross link repair 1C antibody
    • DNA cross link repair 1C protein antibody
    • DNA cross-link repair 1C protein antibody
    • FLJ11360 antibody
    • FLJ36438 antibody
    • hSNM1C antibody
    • OTTHUMP00000045150 antibody
    • Protein A-SCID antibody
    • Protein ARTEMIS antibody
    • PSO2 homolog antibody
    • RS SCID antibody
    • SCIDA antibody
    • Severe combined immunodeficiency type a antibody
    • SNM1 homolog C antibody
    • SNM1 like protein antibody
    • SNM1-like protein antibody
    • SNM1C antibody
    see all

Images

  • All lanes : Anti-Artemis antibody (ab3834) at 2.4 µg/ml

    Lane 1 : AT7BI whole cell lysate
    Lane 2 : MO59K whole cell lysate
    Lane 3 : MO59J whole cell lysate
    Lane 4 : HeLa whole cell lysate
    Lane 5 : CJ whole cell lysate

    Predicted band size: 78 kDa
    Observed band size: 90 kDa
    why is the actual band size different from the predicted?



    Western blot using ab3834 at 1/500 on various cell lysates. CJ is an artemis deficient cell line and so no band is visible.

    CJ is an Artemis deficient cell line and so no band is visible.

  • IHC-P of human spleen using ab3834 at a 1:1000 dilution.  Positive staining of T cells and B lymphocytes is observed.

References

This product has been referenced in:
  • Newman EA  et al. Alternative NHEJ Pathway Components Are Therapeutic Targets in High-Risk Neuroblastoma. Mol Cancer Res 13:470-82 (2015). Read more (PubMed: 25563294) »
  • Sallmyr A  et al. Up-regulation of WRN and DNA ligase III{alpha} in chronic myeloid leukemia: consequences for the repair of DNA double strand breaks. Blood : (2008). Read more (PubMed: 18524993) »
See all 2 Publications for this product

Customer reviews and Q&As

Answer

Thank you for your enquiry. We don't sell the CJ cell line; the Western blot image located on the online datasheet was provided for us by Dr Penny Jeggo, University of Sussex. As you seem to be experiencing some difficulty with this antibody, I would like to try to help you out. Could we get a detailed protocol from you, please? We have some general questions, the answers to which will enable us to investigate this matter as quickly as possible: 1. Please describe the problem (high background, wrong band size, more bands, no band etc). 2. On what material are you testing the antibody in WB? •Species? •Cell extract/ Nuclear extract? •Purified protein? •Recombinant protein? 3. How much protein did you load? •How did you prepare the lysate for the analysis (protease inhibitors etc)? •Did you heat the samples? 4. Primary Antibody •Specification (in which species was it raised against)? •At what dilution(s) have you tested this antibody? •Incubation time, wash step? 5. Secondary Antibody •Specification (in which species was it raised against)? •At what dilution(s) have you tested this antibody? •Incubation time, wash step? •Do you know whether the problems you are experiencing come from the secondary? 6. What detection method are you using? 7. Background bands •Have you eliminated the possibility that any background bands could be due to the secondary antibody? (Run a “No primary” control) •Is the blocking step sufficient? (We recommend blocking the membrane by adding 20 ml of blocking buffer (5% non-fat dry milk, 0.1% Tween-20 in TBS). Incubate for 2 h at room temperature or overnight at 4°C with agitation) •Are your washing steps sufficiently stringent? (Multiple short washes are more effective than fewer longer wash steps) •At what size are the bands migrating? Could they be degradation products of your target? •Please provide an image of your blot (as an e-mail attachment, a faxed image is not sufficient) 8. Optimization attempts •How many times have you tried the Western? •Do you obtain the same results every time e.g. are background bands always in the same place? •What steps have you altered? 9. Did you apply positive and negative controls along with the samples? Please specify.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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