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Index / Trevigen / Anti_Cleaved_Caspase_3 PC /Product Detail : 2305-PC-100 Anti_Cleaved_Caspase_3 PC
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#2305-PC-100 Anti_Cleaved_Caspase_3 PC

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  Price : 483   EUR
548   USD
375   GBP
2031   Zloty
64699   JPY
3729   NOK
3995   SEK
546   CHF

Product name : Anti_Cleaved_Caspase_3 PC

Catalog number : 2305-PC-100

Quantity: 100 µl

Availability: Yes

Supplier name : Trevigen

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More Details about

Anti-Cleaved Caspase-3 Rabbit Polyclonal Antibody


Catalog #: 2305-PC-020

Size: 20 μl

Description: Rabbits were immunized with the KLH-coupled synthetic peptide CRGTELDCGIETD. The polyclonal antibody was affinity purified on a column derivatized with the peptide. It detects the processed large subunit (p18 fragment) but not the unprocessed, zymogen form of caspase-3. In apoptotic lysates, the antibody may recognize the induction of a strong p30 band (Fig. 1).


Physical State: This antibody is provided as affinity purified IgG fraction in 1X PBS without preservative. The final antibody concentration is 0.1 mg/ml.


Immunogen: 13-amino acid peptide sequence from carboxyl terminus of the human and mouse caspase-3 p18 subunit.


Specificity: The antibody detects human and mouse cleaved caspase-3.


Storage: The antibody can be stored at 4°C for at least 1 month, or can be aliquoted and stored frozen at -20°C to -80°C. Avoid repeated freezing and thawing by aliquoting into smaller portions.


Applications: For immunodetection, an antibody dilution of 1:500-1:1000 is recommended. May also be used for immunoprecipitation and immunocytochemistry at a 1:100-1:1000 dilution. Empirical determination will be required for optimal results.


Reference: 1. Srinivasan, A., K.A. Roth, R.O. Sayers, K.S. Shindler, A.M. Wong, L.C. Fritz, and K.J. Tomaselli. 1998. In situ immunodetection of activated caspase-3 in apoptotic neurons in the developing nervous system. Cell Death Diff 5:1004-1016.


Fig 1. Immunoblot of SDS-extracts from mouse cells treated with 25 μM etoposide. Samples were electrophoresed on a 12% Tris-Glycine gel and blotted onto a PVDF membrane. The 18 kDa cleaved caspase-3 was detected by Trevigen’s anti-cleaved caspase-3 followed by anti-rabbit conjugated to horseradish peroxidase and chemiluminescence.

 


Fig. 2. Jurkat cells were treated with 25 μM etoposide for 5 hours to induce apoptosis. Cells were labeled with 1:250 dilution of anti-cleaved caspase-3 followed by anti-rabbit horseradish peroxidase. Detection was performed with Trevigen’s Blue Label and counterstained with Nuclear Fast Red.


Cell Lysates for Western Blotting:


To prepare total cell lysates, cells are solubilized in 1X SDS gel sample buffer (20 mM dithiothiothreitol, 6% SDS, 250 mM Tris, pH 6.8, 10% glycerol, and bromophenol blue) at 5 x 105 - 1 x 106 cells per ml. The extracts are heated in a boiling water bath for 5 minutes. Electrophorese on 12% Tris-Glycine SDS-PAGE gels.


Procedure for Immunoblotting using Peroxidase Detection:


Blotting buffer: 12 mM Tris base, 96 mM Glycine, and 20% MeOH.
Blocking solution: 5% (w/v) nonfat dry milk in PBS.
Antibody solution: 5% (w/v) nonfat dry milk, 0.05% Tween in PBS.


Transfer the electrophoresed proteins to a PVDF membrane and incubate the membrane for 1/2 hour at room temperature in blocking solution. Incubate the membrane overnight at 4°C in antibody solution containing a 1:500 -1:1000 dilution of cleaved caspase-3 rabbit polyclonal antibody. Empirical determination of primary antibody concentration will be required for optimal results. Wash the membrane at room temperature for 15 minutes with 3 changes of 0.05% Tween in PBS. Changing membrane containers often reduces background. Incubate the membrane at room temperature for 1 hour in an antibody solution containing anti-rabbit IgG conjugated to horseradish peroxidase. Empirical determination of secondary antibody concentration will be required for optimal results. Wash the membrane for 15 minutes with 3 changes of 0.05% Tween in PBS. Develop peroxidase reaction using e.g. chemiluminescence.

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Pathways :
WP1023: mRNA processing
WP1027: Glycolysis and Gluconeogenesis
WP1034: Acetylcholine Synthesis
WP1035: Mismatch repair
WP1058: Senescence and Autophagy
WP1078: G1 to S cell cycle control
WP1083: Cell cycle
WP1101: DNA Replication
WP1105: Adipogenesis
WP112: Principle Pathways of Carbon Metabolism
WP1142: mRNA processing
WP1146: Glycolysis and Gluconeogenesis
WP1152: Mismatch repair
WP1174: Senescence and Autophagy
WP1195: G1 to S cell cycle control
WP1200: Cell cycle
WP1204: Mismatch repair
WP1229: mRNA processing
WP123: mRNA processing
WP1232: Notch Signaling Pathway
WP1233: Mismatch repair
WP1257: Mismatch repair
WP1267: Senescence and Autophagy
WP1295: Mismatch repair
WP1305: Senescence and Autophagy

Related Genes :
[Casp4 Casp11 Caspl Ich3] Caspase-4 (CASP-4) (EC 3.4.22.64) (Caspase-11) (CASP-11) (Protease ICH-3) [Cleaved into: Caspase-4 subunit p10; Caspase-4 subunit p20]
[Casp7 Lice2 Mch3] Caspase-7 (CASP-7) (EC 3.4.22.60) (Apoptotic protease Mch-3) (Cysteine protease LICE2) [Cleaved into: Caspase-7 subunit p20; Caspase-7 subunit p11]
[Casp3 Cpp32] Caspase-3 (CASP-3) (EC 3.4.22.56) (Apopain) (Cysteine protease CPP32) (CPP-32) (LICE) (Protein Yama) (SREBP cleavage activity 1) (SCA-1) [Cleaved into: Caspase-3 subunit p17; Caspase-3 subunit p12]
[Casp2 Ich1 Nedd-2 Nedd2] Caspase-2 (CASP-2) (EC 3.4.22.55) (Neural precursor cell expressed developmentally down-regulated protein 2) (NEDD-2) (Protease ICH-1) [Cleaved into: Caspase-2 subunit p18; Caspase-2 subunit p13; Caspase-2 subunit p12]
[CASP8 MCH5] Caspase-8 (CASP-8) (EC 3.4.22.61) (Apoptotic cysteine protease) (Apoptotic protease Mch-5) (CAP4) (FADD-homologous ICE/ced-3-like protease) (FADD-like ICE) (FLICE) (ICE-like apoptotic protease 5) (MORT1-associated ced-3 homolog) (MACH) [Cleaved into: Caspase-8 subunit p18; Caspase-8 subunit p10]
[Casp6] Caspase-6 (CASP-6) (EC 3.4.22.59) (Apoptotic protease Mch-2) [Cleaved into: Caspase-6 subunit p18; Caspase-6 subunit p11]
[Casp1 Il1bc] Caspase-1 (CASP-1) (EC 3.4.22.36) (Interleukin-1 beta convertase) (IL-1BC) (Interleukin-1 beta-converting enzyme) (ICE) (IL-1 beta-converting enzyme) (p45) [Cleaved into: Caspase-1 subunit p20; Caspase-1 subunit p10]
[CASP7 MCH3] Caspase-7 (CASP-7) (EC 3.4.22.60) (Apoptotic protease Mch-3) (CMH-1) (ICE-like apoptotic protease 3) (ICE-LAP3) [Cleaved into: Caspase-7 subunit p20; Caspase-7 subunit p11]
[CASP3 CPP32] Caspase-3 (CASP-3) (EC 3.4.22.56) (Apopain) (Cysteine protease CPP32) (CPP-32) (Protein Yama) (SREBP cleavage activity 1) (SCA-1) [Cleaved into: Caspase-3 subunit p17; Caspase-3 subunit p12]
[Casp3 Cpp32] Caspase-3 (CASP-3) (EC 3.4.22.56) (Apopain) (Cysteine protease CPP32) (CPP-32) (IRP) (LICE) (Protein Yama) (SREBP cleavage activity 1) (SCA-1) [Cleaved into: Caspase-3 subunit p17; Caspase-3 subunit p12]
[CASP3] Caspase-3 (CASP-3) (EC 3.4.22.56) [Cleaved into: Caspase-3 subunit p17; Caspase-3 subunit p12]
[Dredd DCP2 CG7486] Caspase-8 (EC 3.4.22.61) (Death-related ced-3/NEDD2-like protein) [Cleaved into: Caspase-8 subunit p15; Caspase-8 subunit p10]
[CASP9 MCH6] Caspase-9 (CASP-9) (EC 3.4.22.62) (Apoptotic protease Mch-6) (Apoptotic protease-activating factor 3) (APAF-3) (ICE-like apoptotic protease 6) (ICE-LAP6) [Cleaved into: Caspase-9 subunit p35; Caspase-9 subunit p10]
[CASP2 ICH1 NEDD2] Caspase-2 (CASP-2) (EC 3.4.22.55) (Neural precursor cell expressed developmentally down-regulated protein 2) (NEDD-2) (Protease ICH-1) [Cleaved into: Caspase-2 subunit p18; Caspase-2 subunit p13; Caspase-2 subunit p12]
[CASP3] Caspase-3 (CASP-3) (EC 3.4.22.56) [Cleaved into: Caspase-3 subunit p17; Caspase-3 subunit p12]
[Casp9 Mch6] Caspase-9 (CASP-9) (EC 3.4.22.62) (Apoptotic protease Mch-6) (Apoptotic protease-activating factor 3) (APAF-3) (ICE-like apoptotic protease 6) (ICE-LAP6) [Cleaved into: Caspase-9 subunit p35; Caspase-9 subunit p10]
[CASP5 ICH3] Caspase-5 (CASP-5) (EC 3.4.22.58) (ICE(rel)-III) (Protease ICH-3) (Protease TY) [Cleaved into: Caspase-5 subunit p20; Caspase-5 subunit p10]
[CASP4 ICH2] Caspase-4 (CASP-4) (EC 3.4.22.57) (ICE and Ced-3 homolog 2) (ICH-2) (ICE(rel)-II) (Mih1) (Protease TX) [Cleaved into: Caspase-4 subunit 1; Caspase-4 subunit 2]
[Casp2 Ich1] Caspase-2 (CASP-2) (EC 3.4.22.55) (Protease ICH-1) [Cleaved into: Caspase-2 subunit p18; Caspase-2 subunit p13; Caspase-2 subunit p12]
[BTG2 PC3] Protein BTG2 (BTG family member 2) (NGF-inducible anti-proliferative protein PC3)
[CASP10 MCH4] Caspase-10 (CASP-10) (EC 3.4.22.63) (Apoptotic protease Mch-4) (FAS-associated death domain protein interleukin-1B-converting enzyme 2) (FLICE2) (ICE-like apoptotic protease 4) [Cleaved into: Caspase-10 subunit p23/17; Caspase-10 subunit p12]
[CASP6 MCH2] Caspase-6 (CASP-6) (EC 3.4.22.59) (Apoptotic protease Mch-2) [Cleaved into: Caspase-6 subunit p18; Caspase-6 subunit p11]
[CASP1 IL1BC IL1BCE] Caspase-1 (CASP-1) (EC 3.4.22.36) (Interleukin-1 beta convertase) (IL-1BC) (Interleukin-1 beta-converting enzyme) (ICE) (IL-1 beta-converting enzyme) (p45) [Cleaved into: Caspase-1 subunit p20; Caspase-1 subunit p10]
[Drice ICE CG7788] Caspase (EC 3.4.22.-) (drICE) [Cleaved into: Caspase subunit p21; Caspase subunit p12]
[Casp8] Caspase-8 (CASP-8) (EC 3.4.22.61) [Cleaved into: Caspase-8 subunit p18; Caspase-8 subunit p10]
[Btg2 Pc3] Protein BTG2 (BTG family member 2) (NGF-inducible anti-proliferative protein PC3)
[Casp8] Caspase-8 (CASP-8) (EC 3.4.22.61) [Cleaved into: Caspase-8 subunit p18; Caspase-8 subunit p10]
[Dcp-1 CG5370] Caspase-1 (EC 3.4.22.-) [Cleaved into: Caspase-1 subunit p22; Caspase-1 subunit p13]
[Casp6 Mch2] Caspase-6 (CASP-6) (EC 3.4.22.59) (Apoptotic protease Mch-2) [Cleaved into: Caspase-6 subunit p18; Caspase-6 subunit p11]
[Casp1 Il1bc Il1bce] Caspase-1 (CASP-1) (EC 3.4.22.36) (Interleukin-1 beta convertase) (IL-1BC) (Interleukin-1 beta-converting enzyme) (ICE) (IL-1 beta-converting enzyme) (p45) [Cleaved into: Caspase-1 subunit p20; Caspase-1 subunit p10]

Bibliography :
[19508825] Attenuation of ischemia-induced rat brain injury by 2-(-2-benzofuranyl)-2-imidazoline, a high selectivity ligand for imidazoline I(2) receptors.