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|Antigen||Caspase 9, Apoptosis-Related Cysteine Peptidase (CASP9) Antibodies|
|Conjugate||This Caspase 9 antibody is un-conjugated Alternatives|
Immunocytochemistry (ICC), Immunofluorescence (IF), Immunohistochemistry (IHC), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Western Blotting (WB)
|5 references available|
|Supplier||Log in to see|
Product Details anti-Caspase 9 AntibodyTarget Details Caspase 9 Application Details Handling ProductDetails: References for anti-Caspase 9 antibody (ABIN153015) Images
|Immunogen||Recombinant human Caspase 9 pro-domain protein. [UniProt# P55211]|
Target Details Caspase 9Product Details anti-Caspase 9 Antibody Application Details Handling ProductDetails: References for anti-Caspase 9 antibody (ABIN153015) Images back to top
|Alternative Name||Caspase-9 (CASP9 Antibody Abstract)|
|Background||Gene Symbol: CASP9|
|Research Area||Apoptosis/Necrosis, Cancer|
|Pathways||MAPK Signaling, RTK Signaling, Apoptosis, Caspase Cascade in Apoptosis, Fc-epsilon Receptor Signaling Pathway, EGFR Signaling Pathway, Neurotrophin Signaling Pathway, Positive Regulation of Endopeptidase Activity|
Application DetailsProduct Details anti-Caspase 9 Antibody Target Details Caspase 9 Handling ProductDetails: References for anti-Caspase 9 antibody (ABIN153015) Images back to top
|Application Notes||Western Blot 1:500-1:1000, Immunohistochemistry, Immunocytochemistry/Immunofluorescence 1:50-1:250, Immunohistochemistry-Paraffin 1:50-1:250This Caspase 9 (LAP6 96-2-22) antibody is useful for Immunocytochemistry/Immunofluorescence and Western Blot, where bands at ~46 kDa-48 kDa and 35 kDa are seen representing the pro and active forms of Caspase 9, respectively. In ICC/IF cytoplasmic staining can be seen in HeLa cells. Paraffin data from customer review.|
The antibodies are intended for use in vitro experiments only. Our antibodies have not been tested nor are recommended for use in vivo.
Western protocol specific for Caspase 9 Antibody Procedure Guide for NB 500-209 Monoclonal anti-Caspase 9
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.
|Restrictions||For Research Use only|
HandlingProduct Details anti-Caspase 9 Antibody Target Details Caspase 9 Application Details ProductDetails: References for anti-Caspase 9 antibody (ABIN153015) Images back to top
Buffer contains: 0.1 % Sodium Azide
|Precaution of Use||This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.|
|Handling Advice||Avoid freeze-thaw cycles|
|Storage||-80 °C,-20 °C|
|Storage Comment||Aliquot and store at -20°C or -80°C. Avoid freeze-thaw cycles.|
ProductDetails: References for anti-Caspase 9 antibody (ABIN153015)Product Details anti-Caspase 9 Antibody Target Details Caspase 9 Application Details Handling Images back to top
|Product cited in:||
Hsuan, Chyau, Hung, Chen, Chou: "The induction of apoptosis and autophagy by Wasabia japonica extract in colon cancer." in: European journal of nutrition, Vol. 55, Issue 2, pp. 491-503, 2016 (Sample species: Human). Further details: Western Blotting
Kim, Lee, Park, Chung, Im, Noh, Han, Moon, Jung, Ryu: "Expression of SIRT1 and apoptosis-related proteins is predictive for lymph node metastasis and disease-free survival in luminal A breast cancer." in: Virchows Archiv : an international journal of pathology, Vol. 467, Issue 5, pp. 563-70, 2015 (Sample species: Human). Further details: Immunohistochemistry (Paraffin-embedded Sections)
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 (Sample species: Human). Further details: Western Blotting
Kirsch, Doseff, Chau, Lim, de Souza-Pinto, Hansford, Kastan, Lazebnik, Hardwick: "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
Fearnhead, Rodriguez, Govek, Guo, Kobayashi, Hannon, Lazebnik: "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
ImagesProduct Details anti-Caspase 9 Antibody Target Details Caspase 9 Application Details Handling ProductDetails: References for anti-Caspase 9 antibody (ABIN153015) back to top