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Western blot analysis of Caspase-3 was performed by loading 20 ug of U-87 MG (lane 1), Hep G2 (lane 2), Ntera-2 (lane 3), Jurkat (lane 4) and HEK-293 (lane 5) (Fig. A) and Jurkat (lane 1), Jurkat treated for O/N with 3 uM of Staurosporine (lane 2), HeLa (lane 3), HeLa treated for O/N with 1 uM of Etoposide (lane 4) and HeLa treated for O/N with 3 uM of Staurosporine in (lane 5) (Fig. B) cell lysates using Novex® NuPAGE® 4-12 % Bis-Tris gel (NP0322BOX), XCell SureLock™ Electrophoresis System (EI0002), Novex® Sharp Pre-Stained Protein Standard (LC5800), and iBlot® Dry Blotting System (IB21001). Proteins were transferred to a nitrocellulose membrane and blocked with 5 % skim milk for 1 hour at room temperature. Caspase-3 was detected at ~32 kDa using Caspase-3 Mouse Monoclonal Antibody (351600Z) at 1-2 µg/mL in 5 % skim milk at 4°C overnight on a rocking platform. Goat Anti-Mouse - HRP Secondary Antibody (626520) at 1:4000 dilution was used and chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (WP20005). (Fig. B) Upon treatment with Staurosporine/Etoposide there was a reduction in total Caspase-3.
|Tested species reactivity||Human|
|Published species reactivity||Human|
|Host / Isotype||Mouse / IgG2a|
|Immunogen||Full length, recombinant human caspase 3 protein|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Storage Conditions||Maintain refrigerated at 2-8°C for up to 1 month. For long term storage store at -20°C|
|Tested Applications||Dilution *|
|Immunohistochemistry (IHC)||1-2 µg/ml|
|Western Blot (WB)||1-2 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Western Blot (WB)||See 3 publications below|
Caspase 3 is ubiquitously expressed and like other caspases is synthesized as an inactive (32kDa) proenzyme. Upon activation, Caspase 3 is cleaved at Asp28-Ser29 and Asp175-Ser176 thereby generating two subunits of 17kDa and 12kDa, respectively. Recent studies have implicated that Caspase 3 is associated with the induction of apoptosis. Activation of Caspase 3 occurs in response to variety of apoptotic inducers including Fas mediated apoptosis.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
In vitro cytotoxicity of 4'-OH-tamoxifen and estradiol in human endometrial adenocarcinoma cells HEC-1A and HEC-1B.
35-1600Z was used in western blot to assess the effects of 4'-hydroxy-tamoxifen and estradiol on two human endometrial adenocarcinoma cell lines.
|Cuevas ME,Lindeman TE||Oncology reports (33:464)||2015|
|Human||Not Cited||The effect of Longan seed polyphenols on colorectal carcinoma cells.||Chung YC,Lin CC,Chou CC,Hsu CP||European journal of clinical investigation (40:713)||2010|
|Human||Not Cited||Mechanisms of grape seed procyanidin-induced apoptosis in colorectal carcinoma cells.||Hsu CP,Lin YH,Chou CC,Zhou SP,Hsu YC,Liu CL,Ku FM,Chung YC||Anticancer research (29:283)||2009|
apopain; Apopain precursor; CASP-3; caspase 3, apoptosis-related cysteine peptidase; caspase 3, apoptosis-related cysteine protease; CPP-32; CPP32; cysteine protease CPP32; EC 3.4.22.-; ICE3; PARP cleavage protease; procaspase3; protein Yama; SCA-1; SREBP cleavage activity 1; Yama protein
CASP3; CPP32; CPP32B; SCA-1