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Extra Spindle Poles Like 1 (ESPL1) (AA 1866-1996), (C-Term) Antikörper

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Antigen
Epitop
C-Term, AA 1866-1996
(18), (16), (15), (15), (13), (10), (8), (8), (8), (6), (6), (6), (6), (5), (4), (3), (3), (3), (2), (2), (2), (1), (1), (1), (1)
Reaktivität
Human
(184), (40), (28), (2)
Wirt
Maus
(156), (28)
Klonalität (Klon)
Monoklonal ()
Konjugat
Unkonjugiert
(15), (15), (13), (10), (10), (10), (3), (3), (3), (3), (3), (3), (3), (3), (3), (2), (1), (1)
Applikation
Immunzytochemie (ICC), Immunfluoreszenz (IF)
(90), (89), (64), (33), (26), (21), (20), (9), (8), (3), (2)
Pubmed 4 Publikationen vorhanden
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Menge 0.1 mL
Lieferung nach Deutschland ( )
Verfügbarkeit
Immunogen Maltose-Binding Protein fusion of C-terminal fragment of human Separase (amino acids 1866-1996).
Klon XJ11-4D7
Isotyp IgG2a
Reinigung Ascites
Andere Bezeichnung Separin / ESPL1 (ESPL1 Antibody Abstract)
Hintergrund Separase is a cysteine protease that is essential for mitotic progression by separatingsister chromatids. Each cell must receive one chromatid of every chromosome, duringmitosis. Cohesin plays an important role in cohering sister chromatids during theprophase through anaphase stages of mitosis, making certain that genomic informationis replicated accurately. As the cellular division process continues, separase destroyscohesin by means of cleavage, allowing the chromatids to separate and divide with thecell. Separase activity is highly regulated. It not only cleaves cohesin at the onset ofanaphase but also cleaves itself, promoting downregulation of separase after anaphase. Should a human cell become an aneuploid (one too many or too few chromatids), theembryo most likely will not survive. Should the embryo survive, it will most likelydevelop severe birth defects or later develop malignant cancers.Separase antibodies can be used as a specific marker for centrosomes of mitotic cells. The staining of separase in centrosomes can be detected from prophase of mitosis upuntil anaphase. Alternate Names: anti-Caspase like protein ESPL1 antibody, anti-ESP1 antibody, anti-ESPL1 antibody,anti-Extra spindle poles like 1 antibody, anti-KIAA0165 antibody, anti-Separin antibody,anti-Similar to fission yeast cut1and gene antibody, anti-SSE antibody.
Gene Symbol: ESPL1
Gen-ID 9700
UniProt Q14674
Applikationshinweise This Separase (XJ11-4D7) antibody is useful for Immunocytochemistry/Immunofluorescence. In ICC/IF, this antibody recognizes endogenous Separase in paraformaldehyde-fixated preparations of human cells.
Recommended dilutions: Immunocytochemistry/Immunofluorescence 1:1000
Protokoll Immunofluorescence protocol specific for Separase Antibody Immunofluorescence Procedure Cell Preparation
1. HeLa cells are grown on coverslips and enriched in a fraction of mitotic cells by double thymidine block/release protocol.
. Briefly, cells are allowed to attach to the glass coverslips for 16 hours.
. Thymidine is added to the culturing medium at a final concentration of 2 mM for 18 hours.
. Cells are washed 2 times with PBS.
. Fresh medium without thymidine is added.
. Cells are incubated in the thymidine-free medium for 8 hours.
. Thymidine is added again to a final 2 mM concentration for 18 hours again.
. After the second thymidine incubation, the cells are washed 2 times with PBS.
. Fresh thymidine-free medium is added again. 10 .Starting at 8 hours after the second thymidine removal, cells are analyzed by light microscopy and once population of mitotic cells are about 50 % (usually around 9-10 hours after the second release) the cell staining procedure begins. Cell Staining
. Cells are washed 2 times with Dulbecco's PBS with Ca and Mg (D-PBS) and permeabilized prior to fixation by incubation with 0.05 % Triton X-100 for 1 min.
. Permeabilizing solution is aspirated and 4 % paraformaldehyde is added for 30-45 min.
. Paraformaldehyde is removed by washing the coverslips 3 times with D-PBS.
. Cells are blocked by SuperBlock reagent (Pierce) in PBS supplemented with 0.5 % Triton X-100 for 30 min.
. The coverslips are washed 3 times with 0.05 % Tween-20 in PBS (PBS-T).
. Staining to detect separase is carried out at a dilution of 1:1,000 of mouse monoclonal anti-separase prepared in PBS-T and incubated with the cells on coverslips for 1 hour at room temperature (RT).
. Coverslips are washed 3 x 5 min. washes with PBS-T.
. Incubate cells with an anti-mouse Cy3 conjugated fluorochrome-labeled secondary. The secondary antibody is diluted in PBS-T and incubated for 20 min.
. Coverslips are incubated with DAPI at a final concentration of 10 ng/ml in PBS-T for 5 min.10.Cells are washed 5 x 5 min with PBS-T. 11.Prior to mounting, the coverslips are washed once with PBS without detergents.
. Micrographs are taken by a fluorescent microscope.
Beschränkungen Nur für Forschungszwecke einsetzbar
Format Liquid
Konservierungsmittel Sodium azide
Vorsichtsmaßnahmen WARNING: Reagents contain sodium azide. Sodium azide is very toxic if ingested or inhaled. Avoid contact with skin, eyes, or clothing. Wear eye or face protection when handling. If skin or eye contact occurs, wash with copious amounts of water. If ingested or inhaled, contact a physician immediately. Sodium azide yields toxic hydrazoic acid under acidic conditions. Dilute azide-containing compounds in running water before discarding to avoid accumulation of potentially explosive deposits in lead or copper plumbing.
Handhabung Avoid freeze-thaw cycles
Lagerung -20 °C
Informationen zur Lagerung Aliquot and store at -20 °C or -80 °C.
Bilder des Herstellers
Immunfluoreszenz (IF) image for anti-Extra Spindle Poles Like 1 (ESPL1) (AA 1866-1996), (C-Term) Antikörper (ABIN151796) Overlay [blue] of centrosomal staining in HeLa cells. Centrosomal staining of separas...
Produkt verwendet in: Fujita, Epperly, Zou et al.: "Regulation of the anaphase-promoting complex-separase cascade by transforming growth factor-beta modulates mitotic progression in bone marrow stromal cells." in: Molecular biology of the cell, Vol. 19, Issue 12, pp. 5446-55, 2008 (PubMed).

Rubinek, Chesnokova, Wolf et al.: "Discordant proliferation and differentiation in pituitary tumor-transforming gene-null bone marrow stem cells." in: American journal of physiology. Cell physiology, Vol. 293, Issue 3, pp. C1082-92, 2007 (PubMed).

Chestukhin, Pfeffer, Milligan et al.: "Processing, localization, and requirement of human separase for normal anaphase progression." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 100, Issue 8, pp. 4574-9, 2003 (PubMed).

Allgemeine Veröffentlichungen Chestukhin, DeCaprio: "Western blot screening for monoclonal antibodies against human separase." in: Journal of immunological methods, Vol. 274, Issue 1-2, pp. 105-13, 2003 (PubMed).

Produktnummer ABIN151796

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