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Caspase 9 Antikörper (Caspase 9, Apoptosis-Related Cysteine Peptidase)

Details für Produkt anti-CASP9 Antikörper Nr. ABIN153015, Anbieter: Anmelden zum Anzeigen Neu
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Antigen
Alternativen
Reaktivität
Human
(416), (115), (110), (15), (13), (9), (6), (5), (3), (3), (2), (1), (1), (1)
Wirt
Maus
(425), (48), (1)
Klonalität (Klon)
Monoklonal ()
Konjugat
Dieser Caspase 9 Antikörper ist unkonjugiert
(12), (10), (7), (5), (4), (4), (4), (4), (4), (4), (4), (4), (3), (3), (3)
Applikation
Western Blot (WB), Immunzytochemie (ICC), Immunfluoreszenz (IF)
(395), (163), (120), (80), (69), (61), (46), (38), (13), (12), (8), (6), (2), (2)
Pubmed 3 Publikationen vorhanden
Hersteller Anmelden zum Anzeigen Neu
Produkt-ID vom Hersteller Anmelden zum Anzeigen Neu
Menge 0.2 mL
Lieferung nach Deutschland ( )
Verfügbarkeit
Immunogen Recombinant human Caspase 9 pro-domain protein.
Klon LAP6 96-2-22
Isotyp IgG1 kappa
Reinigung Ascites
Andere Bezeichnung Caspase-9 (CASP9 Antibody Abstract)
Hintergrund Caspases are cysteine proteases, expressed as inactive precursors, that mediateapoptosis by proteolysis of specific substrates. Caspases have the ability to cleave afteraspartic acid residues. There are two classes of caspases involved in apoptosis,initiators (activation by receptor cluster) and effectors (activation by mitochondrialpermeability transition). Proapoptotic signals autocatalytically activate initiatorcaspases, such as Caspase-8 and Caspase-9. Activated initiator caspases thenprocess effector caspases, such as Caspase-3 and Caspase-7, which in turn cause cellcollapse. Alternate Names: anti-APAF 3 antibody, anti-APAF3 antibody, anti-Apoptosis related cysteine peptidaseantibody, anti-Apoptotic protease activating factor 3 antibody, anti-Apoptotic proteaseMch 6 antibody, anti-CASP 9 antibody, anti-ICE LAP6 antibody, anti-ICE like apoptoticprotease 6 antibody.
Gene Symbol: CASP9
Gen-ID 842
UniProt P55211
Forschungsgebiet Apoptosis/Necrosis
Pathways MAPK Signalweg, RTK Signalweg, Apoptose, Caspase Kaskade in der Apoptose, Fc-epsilon Rezeptor Signalübertragung, EGFR Signaling Pathway, Neurotrophin Signalübertragung
Applikationshinweise This Caspase 9 (LAP6 96-2-22) antibody is useful for Immunocytochemistry/Immunofluorescence and Western Blot, where bands at ~46kDa-48kDa and 35kDa are seen representing the pro and active forms of Caspase 9, respectively. In ICC/IF cytoplasmic staining can be seen in HeLa cells.
Recommended dilutions: Immunocytochemistry/Immunofluorescence 1:50-1:250, Western Blot 1:500-1:1000
Protokoll Western protocol specific for Caspase 9 Antibody Procedure Guide for NB 500-209 Monoclonal anti-Caspase 9

Western Protocol

1) Run a 10-*15 % SDS polyacrylamide gel, loading ~20 µg of cell extract per lane.
2) Transfer the proteins to a membrane.
3) Block the membrane in PT-T20 [20 mM Tris-HCl, pH 7.4 / 150 mM NaCl/0.5 % Tween 20] + 5 % NFDM [non-fat dry milk], for 3 hour at room temperature (RT), shaking gently.
4) Rinse the membrane twice with PT-T20.
5) Incubate the membrane in anti-Caspase 9 [NB 500-209], diluted 1:500-1:1,000 in PT-T20 + 5 % NFDM, for 60 minutes at RT.
6) Wash the membrane for 15 minutes, 3 times, in PT-T20 at RT.
7) Incubate the membrane in anti-mouse conjugated to HRP (secondary antibody), diluted in PT-T20 + 5 % NFDM for 60 minutes at RT.
8) Wash the membrane for 15 minutes, 3 times, in PT-T20 at RT.
9) Develop membrane in chemiluminescent reagents, as instructed by kit-vendor.

*Positive control(s): Human kidney 293 cells

To Activate the Cell Extracts:
Bring the extract to a final concentration of 5 mM dATP.
Incubate the extracts at 37?C for 15-30 minutes.Immunocytochemistry/Immunofluorescence Protocol for Caspase 9 Antibody Immunocytochemistry ProtocolCulture cells to appropriate density in 35 mm culture dishes or 6-well plates.
1. Remove culture medium and add 10 % formalin to the dish. Fix at room temperature for 30 minutes.
. Remove the formalin and add ice cold methanol. Incubate for 5-10 minutes.
. Remove methanol and add washing solution (i.e. PBS). Be sure to not let the specimen dry out. Wash three times for 10 minutes.
. To block nonspecific antibody binding incubate in 10 % normal goat serum from 1 hour to overnight at room temperature.
. Add primary antibody at appropriate dilution and incubate at room temperature from 2 hours to overnight at room temperature.
. Remove primary antibody and replace with washing solution. Wash three times for 10 minutes.
. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
. Remove antibody and replace with wash solution, then wash for 10 minutes. Add Hoechst 33258 to wash solution at 1:25,0000 and incubate for 10 minutes. Wash a third time for 10 minutes.
. Cells can be viewed directly after washing. The plates can also be stored in PBS containing Azide covered in Parafilm (TM). Cells can also be cover-slipped using Fluoromount, with appropriate sealing.*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.
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
Western Blot (WB) image for anti-Caspase 9 Antikörper (Caspase 9, Apoptosis-Related Cysteine Peptidase) (ABIN153015) The pro and active forms of Caspase 9 detected in active 293 cell lysate (lane 2). La...
Produkt verwendet in: Martin, Fearnhead: "Apocytochrome c blocks caspase-9 activation and Bax-induced apoptosis." in: The Journal of biological chemistry, Vol. 277, Issue 52, pp. 50834-41, 2002 (PubMed).

Allgemeine Veröffentlichungen Kirsch, Doseff, Chau et al.: "Caspase-3-dependent cleavage of Bcl-2 promotes release of cytochrome c." in: The Journal of biological chemistry, Vol. 274, Issue 30, pp. 21155-61, 1999 (PubMed).

Fearnhead, Rodriguez, Govek et al.: "Oncogene-dependent apoptosis is mediated by caspase-9." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 95, Issue 23, pp. 13664-9, 1998 (PubMed).

Produktnummer ABIN153015

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