Immunofluorescence analysis of Caspase-3 was done on 70% confluent log phase HeLa cells treated with 5 uM of Staurosporine for 16 hours. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with Caspase-3 (9H19L2) ABfinity™ Rabbit Monoclonal antibody (700182) at 1 ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing Cytoplasmic localization. Panel e is untreated cell showing no signal. Panel f is a no primary antibody control. The images were captured at 60X magnification
|Tested species reactivity||Human, Mouse|
|Published species reactivity||Pig, Human, Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Immunogen||A peptide corresponding to amino acids 171-175 of P42574.|
|Contains||0.09% 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 *|
|Flow Cytometry (Flow)||0.5-1 ug/test|
|Immunocytochemistry (ICC)||1-2 µg/ml|
|Immunofluorescence (IF)||1-2 µg/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||1:10-1:50|
|Western Blot (WB)||0.1-0.2 ug/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.
This antibody is predicted to react with bovine, canine, feline, hamster, mouse, primate, pufferfish, rabbit, rat, porcine and Xenopus based on sequence homology.
ABfinity™ recombinant antibodies are rabbit monoclonal antibodies, unmatched for producing superior results. ABfinity™ antibodies are developed by immunizing animals, screening for functionality, cloning the immunogen-specific antibody genes into high-level mammalian expression vectors, produced on a large scale, and purified with Protein A.
ABfinity™ monoclonal antibodies resemble rabbit monoclonals isolated from serum or produced by hybridomas, but demonstrate greater specificity and sensitivity. Because ABfinity™ recombinant antibodies are derived from cloned DNA sequences of the heavy and light antibody chains, they are not susceptible to cell-line drift or lot-to-lot variation, thus allowing for peak specificity and performance.
Intact IgG appears on a non-reducing gel as ~150 kDa band and upon reduction generating a ~25 kDa light chain band and a ~50 kDa heavy chain.
Caspases are a family of cysteine proteases that centrally controls apoptotic machinery. Caspases can be grouped according to their substrate specificities that are largely determined by the amino acids preceding the cleavage site. One group of caspases that include -6, -8, is specific for the substrate V/LEXD. This substrate is a site similar to those found in caspase proenzymes. This group of caspases may function as initiators of a proteolytic cascade by activating pro-caspases to amplify a death signal. A second group of caspases is specific for the substrate DEXD that is related to sites found on target proteins cleaved during apoptosis.
Caspase-3 is a member of the interleukin-1 beta converting enzyme, Caspase-8, nuclear lamins and others. The overexpression of Caspase-3 can result in apoptosis. Likewise, the inhibition of Caspase-3 or other caspases can prevent cells from entering the apoptotic pathway. Recent evidence has revealed a link between plasma caspase-3 and atherosclerosis and it role in activation of apoptosis in breast cancer mediated by siRNA-mediated Apollon silencing. This antibody is specific for the cleaved (active) form of caspase-3.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Comparison of 2, 5, and 20 % O2 on the development of post-thaw human embryos.
700182 was used in immunocytochemistry to characterize the development of post-thaw human embryos by comparing 2,5,a nd 20% O2 levels
|Yang Y,Xu Y,Ding C,Khoudja RY,Lin M,Awonuga AO,Dai J,Puscheck EE,Rappolee DA,Zhou C||Journal of assisted reproduction and genetics (33:919)||2016|
Detection and quantification of farnesol-induced apoptosis in difficult primary cell cultures by TaqMan protein assay.
700182 was used in immunocytochemistry and western blot to develop a TaqMan protein assay of active caspase-3 to quantitate farnesol-induced apoptosis in primary cell cultures
|Pfister C,Pfrommer H,Tatagiba MS,Roser F||Apoptosis : an international journal on programmed cell death (18:452)||2013|
|Not Applicable||Not Cited||
Identification of 1,2,3,4,6-Penta-O-galloyl-ß-d-glucopyranoside as a Glycine N-Methyltransferase Enhancer by High-Throughput Screening of Natural Products Inhibits Hepatocellular Carcinoma.
700182 was used in western blot to employ the method of high-throughput screening of natural products that inhibit hepatocellular carcinoma by identifying 1,2,3,4,6-Penta-O-galloyl-beta-d-glucopyranoside as a glycine N-methyltransferase enhancer
|Kant R,Yen CH,Lu CK,Lin YC,Li JH,Chen YM||International journal of molecular sciences (17:null)||2016|
Investigating the role of shape on the biological impact of gold nanoparticles in vitro.
700182 was used in western blot to assess the impact of gold nanoparticle geometry on the biochemical response of Calu-3 epithelial cells.
|Tian F,Clift MJ,Casey A,Del Pino P,Pelaz B,Conde J,Byrne HJ,Rothen-Rutishauser B,Estrada G,de la Fuente JM,Stoeger T||Nanomedicine (London, England) (10:2643)||2015|
Functional proteomic analysis reveals the involvement of KIAA1199 in breast cancer growth, motility and invasiveness.
700182 was used in immunohistochemistry to study the role of the novel gene KIAA1199 in breast cancer
|Jami MS,Hou J,Liu M,Varney ML,Hassan H,Dong J,Geng L,Wang J,Yu F,Huang X,Peng H,Fu K,Li Y,Singh RK,Ding SJ||BMC cancer (14:null)||2014|
|Not Applicable||Not Cited||
Effect of memantine: A NMDA receptor blocker, on ethambutol-induced retinal injury.
700182 was used in immunohistochemistry - paraffin section to study ethambutol-induced retinal injury and the effect of memantine, an NMDA receptor blocker
|Abdel-Hamid AA,Firgany Ael-D,Ali EM||Annals of anatomy = Anatomischer Anzeiger : official organ of the Anatomische Gesellschaft (204:86)||2016|
Serial diffusion MRI to monitor and model treatment response of the targeted nanotherapy CRLX101.
700182 was used in immunohistochemistry to study the use of serial diffusion MRI to monitor the response to a cyclodextrin polymer nanoparticle containing a DNA topoisomerase I inhibitor in a murine lymphoma model
|Ng TS,Wert D,Sohi H,Procissi D,Colcher D,Raubitschek AA,Jacobs RE||Clinical cancer research : an official journal of the American Association for Cancer Research (19:2518)||2013|
|Not Applicable||Not Cited||
Apoptosis-dependent acute lung injury and repair after intratracheal instillation of noradrenaline in rats.
700182 was used in immunohistochemistry - paraffin section to assess intratracheal instillation of noradrenaline in rats and apoptosis-dependent acute lung injury and repair
|Uhal BD,Rayford H,Zhuang J,Li X,Laukka J,Soledad-Conrad V||Experimental physiology (88:269)||2003|
|Pig||Not Cited||Impact of diet deprivation and subsequent overallowance during gestation on mammary gland development and lactation performance.||Farmer C,Palin MF,Martel-Kennes Y||Journal of animal science (92:141)||2014|