Overview

  • Product name
    Anti-ORC6L antibody [3A4]
    See all ORC6L primary antibodies
  • Description
    Rat monoclonal [3A4] to ORC6L
  • Host species
    Rat
  • Specificity
    ab22536 recognises ORC6L.
  • Tested applications
    Suitable for: IP, WB, ChIPmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Full length protein (Human): ORC6L

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
    Preservative: 0.1% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • Purity
    Protein G purified
  • Clonality
    Monoclonal
  • Clone number
    3A4
  • Myeloma
    x63-Ag8.653
  • Isotype
    IgG2a
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab22536 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
IP Use at an assay dependent dilution.
WB Use at an assay dependent dilution. Predicted molecular weight: 28 kDa.
ChIP Use at an assay dependent dilution.

Target

  • Function
    Component of the origin recognition complex (ORC) that binds origins of replication. DNA-binding is ATP-dependent, however specific DNA sequences that define origins of replication have not been identified so far. ORC is required to assemble the pre-replication complex necessary to initiate DNA replication.
  • Involvement in disease
    Defects in ORC6 are the cause of Meier-Gorlin syndrome type 3 (MGORS3) [MIM:613803]. MGORS3 is a syndrome characterized by bilateral microtia, aplasia/hypoplasia of the patellae, and severe intrauterine and postnatal growth retardation with short stature and poor weight gain. Additional clinical findings include anomalies of cranial sutures, microcephaly, apparently low-set and simple ears, microstomia, full lips, highly arched or cleft palate, micrognathia, genitourinary tract anomalies, and various skeletal anomalies. While almost all cases have primordial dwarfism with substantial prenatal and postnatal growth retardation, not all cases have microcephaly, and microtia and absent/hypoplastic patella are absent in some. Despite the presence of microcephaly, intellect is usually normal.
  • Sequence similarities
    Belongs to the ORC6 family.
  • Post-translational
    modifications
    Phosphorylated upon DNA damage, probably by ATM or ATR.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • ORC 6 antibody
    • Orc6 antibody
    • ORC6_HUMAN antibody
    • ORC6L antibody
    • Origin recognition complex subunit 6 antibody
    • origin recognition complex subunit 6 homolog like antibody
    • origin recognition complex subunit 6 homolog like (yeast) antibody
    • origin recognition complex subunit 6 like (yeast) antibody
    see all

References

ab22536 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Answer

Thank you for your enquiry. At Abcam we qualify all of our "ChIP grade" antisera in two ways. Firstly we operate an in house ChIP testing programme with an in house development scientist. Secondly we collaborate with groups that have extensive experience in ChIP. On this occassion ab22536 was used by and published by: Ritzi M et al. Complex protein-DNA dynamics at the latent origin of DNA replication of Epstein-Barr virus. J Cell Sci 116:3971-84 (2003). PubMed: 12953058 Accoding to the publication ChIP was performed as follows: "ChIP assay and PCR analysis For ChIP experiments, 1´107 nuclei were prepared for each immunoprecipitation as described above. Nuclei were washed at a concentration of 1´108 nuclei ml–1 in ice-cold buffer A supplemented with 200 mM NaCl. After centrifugation (1300 g, 5 minutes, 4°C) nuclei were carefully resuspended in 1 ml buffer A. Then, 9 ml prewarmed buffer A supplemented with 1.1% formaldehyde were added and the nuclei cross-linked for 10 minutes at 37°C. Fixed nuclei were washed twice with PBS with 0.5% NP40, resolved in 2.7 ml LSB (10 mM Hepes pH 7.9, 10 mM KCl, 1.5 mM MgCl2) and lysed by adding 300 ml 20% Sarkosyl. The chromatin was transferred onto a 40 ml sucrose cushion (LSB plus 100 mM sucrose) and centrifuged (10 minutes, 4°C, 4000 g). Supernatant was removed and the chromatin was resuspended in 2 ml TE and sonicated (Branson sonifier 250-D, 35% amplitude, 2 minutes in 1 second intervals). For partial DNA digests, 2 mM CaCl2 and 8 U micrococcal nuclease (MNase) (Roche) were added to the chromatin and incubated for 10 minutes at 37°C. The reaction was stopped by adding 5 mM EGTA. For immunoprecipitation, the extract was adjusted with 1/10 volume of 11´ NET (550 mM Tris-HCl pH 7.4, 1.65 M NaCl, 5.5 mM EDTA, 5.5% NP40). 10 mg affinity-purified polyclonal antibodies (HsOrc3p, HsMcm3/7p), 15 ml of polyclonal HsOrc1p antiserum or 50 ml supernatant of monoclonal antibodies (EBNA1, HsOrc6p) were added respectively. The immunoprecipitation and purification of co-precipitated DNA was performed as illustrated (Schepers et al., 2001). Real-time PCR analysis was performed according to the manufacturers instructions using the same parameters and primer pairs as described (Schepers et al., 2001). A detailed protocol for the ChIP experiments are available (http://haema145.gsf.de/)". This protocol does not deviate significantly from Abcams recommended ChIP protocol aside from a sucrose nuclei purification and an MNase digestion. I hope this information helps. Please do not hesiate to contact me should you require further assistance.

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