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ESPL1 Antikörper (Extra Spindle Poles Like 1) (C-Term)

Details für Produkt anti-ESPL1 Antikörper Nr. ABIN151796, Anbieter: Anmelden zum Anzeigen
Antigen
  • AL024103
  • AU045071
  • Cerp
  • ESP1
  • mp:zf637-3-001691
  • PRCE
  • SEPA
  • SSE
Epitop
C-Term
20
20
18
16
14
10
9
8
8
8
7
6
6
6
6
4
3
3
3
2
2
2
2
1
1
1
Reaktivität
Human
195
41
29
1
Wirt
Maus
162
33
Klonalität (Klon)
Monoklonal ()
Konjugat
Dieser ESPL1 Antikörper ist unkonjugiert
15
15
15
10
10
10
5
4
4
3
3
3
3
3
3
2
2
2
2
1
1
Applikation
Immunocytochemistry (ICC), Immunofluorescence (IF)
94
90
71
35
25
21
20
7
6
4
2
Hersteller
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Immunogen Maltose-Binding Protein fusion of C-terminal fragment of human Separase (amino acids 1866-1996). [UniProt# Q14674]
Klon XJ11-4D7
Isotyp IgG2a
Reinigung Ascites
Andere Bezeichnung Separase (ESPL1 Antibody Abstract)
Hintergrund Gene Symbol: ESPL1
UniProt Q14674
Forschungsgebiet Cell Cycle, Chromatin and Nuclear Signaling
Applikationshinweise Immunocytochemistry/Immunofluorescence 1:1000
Kommentare

The antibodies are intended for use in vitro experiments only. Our antibodies have not been tested nor are recommended for use in vivo.

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
Buffer Buffer contains: 0.1 % Sodium Azide
Konservierungsmittel Sodium azide
Vorsichtsmaßnahmen This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handhabung Avoid freeze-thaw cycles
Lagerung -20 °C,-80 °C
Informationen zur Lagerung Aliquot and store at -20°C or -80°C. Avoid freeze-thaw cycles.
Bilder des Herstellers
Immunofluorescence (IF) image for anti-ESPL1 Antikörper (Extra Spindle Poles Like 1) (C-Term) (ABIN151796) Overlay [blue] of centrosomal staining in HeLa cells. Centrosomal staining of separas...
Produkt verwendet in: Kim, Jeon, Ha, Park, Kim, Shin, Lee, Chung, Lee: "Functional interaction between BubR1 and securin in an anaphase-promoting complex/cyclosomeCdc20-independent manner." in: Cancer research, Vol. 69, Issue 1, pp. 27-36, 2009 (PubMed).

Fujita, Epperly, Zou, Greenberger, Wan: "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).

Sak, Fegers, Groneberg, Stuschke: "Effect of separase depletion on ionizing radiation-induced cell cycle checkpoints and survival in human lung cancer cell lines." in: Cell proliferation, Vol. 41, Issue 4, pp. 660-70, 2008 (PubMed). (Probematerial (Species): Human).

Rubinek, Chesnokova, Wolf, Wawrowsky, Vlotides, Melmed: "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, DeCaprio, Pellman: "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).