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Monoclonal Antibodies

Anti-BORIS [20B11]

Invented by Prof Elena Klenova from University of Essex
Invented at University of Essex
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Monoclonal anti- BORIS C-terminal antibody from clone 20B11 specifically recognises the C terminal domain of BORIS, and the endogenous and exogenous BORIS protein. Cell lysates were resolved by SDS-PAGE, blotted and probed with the original mouse supernatant of the 20B11 clone. Positions of BORIS and the C-terminal domain of BORIS are indicated. B, bacterially expressed BORIS; *, 293T cells transfected with 0.5μg of the plasmid expressing BORIS; M, Molecular marker.
Immunocytochemical staining of MCF7 cells with the mouse anti-BORIS C-terminal antibody, 20B11. Cells were immunostained with the supernatants from the clone 20B11 at following dilutions: (A) 20B11 undiluted supernatant and (B) 20B11 at 1:25. (B) Cells have been counterstained with the nuclear marker hematoxylin (blue). Images were taken at x60 magnification using the Olympus BX41 microscope. Expression of BORIS protein is indicated by the presence of brown staining, with a pattern characteristic for BORIS ( both, cytoplasmic and nuclear).
Immunoprecipitation assay using the anti-BORIS 20B11 antibody The 83-kDa protein can be immunoprecipitated with the mouse anti- BORIS C terminal specific antibody. Proteins were immunoprecipitated from the MCF7 cell extracts with the mouse anti- BORIS C terminal antibody 20B11 in their original supernatant form using the following volumes: (1) 400 μl of 20B11; (2) 800 μl of 20B11; (3) Input; (4) MCF7 cell lysates as BORIS positive control. The extracts from MCF7 cells were incubated overnight at 40C followed by 2 hour incubation with protein G sepharose beads. Proteins were resolved by SDS-PAGE, and Western blot analysis was performed with the concentrated form of the mouse anti- BORIS N terminal (4A7) specific antibody (1:500).
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Catalogue Number 152812
Applications ELISA IHC IF IP WB
Antigen/Gene or Protein Targets BORIS
Synonyms Brother of the regulator of imprinted sites, Cancer/testis antigen 27, CCCTC binding factor (zinc finger protein) like, CCCTC-binding factor, CT27, CTCF paralog, CTCF T, CTCF-like protein, Ctcfl, CTCFL_HUMAN, dJ579F20.2, HMG 1L1, HMGB1L1, MGC163358, MGC169105, MGC169106
Reactivity Human
Relevance Reagent for research, diagnostic tool. BORIS protein has been identified as Cancer-Testis Antigen (CTA) with testis-specific paralogue of the CCCTC-binding factor. Recent studies have demonstrated that d BORIS is directly responsible for the transcriptional activation of TSP50 (testes-specific protease 50).
Host Mouse
Immunogen Synthetic peptide within the BORIS C-terminal domain (aa 614-648) (CG)GEMFPVACRETTARVKEE (NB - the first two aa do not belong to BORIS)
Positive Control MCF7 cell lysates
Subclass IgG3 kappa
Molecular Weight (kDa) 83
Recommended Growing Conditions Vials with hybridoma cell from the liquid nitrogen storage should be rapidly thawed. After thawing, the cell suspension needs to be added slowly using a sterile pipette into 10ml of warmed HM20 medium, the suspended cells centrifuged at 1200 rpm for 10 min and the supernatant discarded. The cell pellet should be gently re-suspended in 9ml of HM20-10% HCF (Hybridoma Cloning Factor, BioVeris Corporation) and transferred into a small flask (T25). Cells should be incubated at 37°C and 10% CO2 and passaged when they were 90% confluent. Cells should be successively grown in the medium with gradually decreasing amounts of HCF from 10% to 5% to 2.5% and finally without any HCF. Depending upon the confluency of the culture, cells should be transferred from the small flask to a medium sized flask (T75) during one of these stages.
Research Area Cancer, Epigenetics & Nuclear Signalling

References

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References: 1 entry

Debruyne et al. 2019. Nature. 572(7771):676-680. PMID: 31391581.

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References: 1 entry

Debruyne et al. 2019. Nature. 572(7771):676-680. PMID: 31391581.

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