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Rabbit Anti-Caspase-3 antibody (bs-0081R)
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说明书: 50ul  100ul  200ul
20ul/380.00元
50ul/780.00元
100ul/1380.00元
200ul/2200.00元
大包装/询价

产品编号 bs-0081R
英文名称 Caspase-3
中文名称 活化半胱胺酸蛋白酶蛋白-3抗体
别    名 Caspase-3 subunit p17; APOPAIN; CASP3; Caspase 3 apoptosis related cysteine protease; Caspase3; CPP32; CPP32B; Cysteine protease CPP32; Human cysteine protease CPP32 isoform alpha mRNA complete cds; PARP cleavage protease; SCA 1; SCA1; SREBP cleavage activity 1; Yama; CASP3_HUMAN; Caspase-3; CASP-3; Apopain; Protein Yama; SREBP cleavage activity 1; SCA-1.  
Specific References  (63)     |     bs-0081R has been referenced in 63 publications.
[IF=3.47] Kubatka, Peter, et al. "Oregano demonstrates distinct tumour-suppressive effects in the breast carcinoma model." European Journal of Nutrition (2016): 1-14.  IHC-P ;  Rat.  
[IF=1.06] Zhang, Xiaolin, and Hao Yu. "Matrine inhibits diethylnitrosamine-induced HCC proliferation in rats through inducing apoptosis via p53, Bax-dependent caspase-3 activation pathway and down-regulating MLCK overexpression (Supplement 2016)." Iranian Journal of Pharmaceutical Research (2016).  WB ;  Rat.  
[IF=3.40] Gao, Hui, et al. "Hispidulin induces mitochondrial apoptosis in acute myeloid leukemia cells by targeting extracellular matrix metalloproteinase inducer."American Journal of Translational Research 8.2 (2016): 1115-1132  WB ;  Human.  
[IF=2.53] Yoshinaga, Ayana, et al. "NEU3 inhibitory effect of naringin suppresses cancer cell growth by attenuation of EGFR signaling through GM3 ganglioside accumulation." European Journal of Pharmacology (2016).  WB ;  Human.  
[IF=0.00] Petty, Howard R. "NANOPARTICLE THERAPY IN CANCER." U.S. Patent No. 20,150,335,744. 26 Nov. 2015.  IF(ICC) ;  
[IF=2.90] Shan, Ming, and Ting-Jun Fan. "Cytotoxicity of carteolol to human corneal epithelial cells by inducing apoptosis via triggering the Bcl-2 family protein-mediated mitochondrial pro-apoptotic pathway." Toxicology in Vitro (2016).  ELISA ;  Human.  
[IF=2.10] Hu, Hanhua, et al. "Drug-induced apoptosis of Echinococcus granulosus protoscoleces." Parasitology research 109.2 (2011): 453-459.  IHC-P ;  Others.  
[IF=1.51] Sun, Y‐L., et al. "Relationship between apoptosis and proliferation in granulosa and theca cells of cystic follicles in sows." Reproduction in domestic animals 47.4 (2012): 601-608.  IHC-P ;  Pig.  
[IF=1.55] Ding, Qing, et al. "8‑bromo‑7‑methoxychrysin induces apoptosis by regulating Akt/FOXO3a pathway in cisplatin‑sensitive and resistant ovarian cancer cells." Molecular medicine reports 12.4 (2015): 5100-5108.  WB ;  Human.  
[IF=4.38] Ma, Xiangyi, et al. "DT390-triTMTP1, a novel fusion protein of diphtheria toxin with tandem repeat TMTP1 peptide, preferentially targets metastatic tumors." Molecular pharmaceutics 10.1 (2013): 115-126.  IHC-P ;  Human.  
[IF=1.55] Han, Ning, et al. "Agmatine protects Müller cells from high‑concentration glucose‑induced cell damage via N‑methyl‑D‑aspartic acid receptor inhibition." Molecular medicine reports 12.1 (2015): 1098-1106.  WB ;  Rat.  
[IF=2.86] Liu, Yaping, et al. "Overexpression of PRL7D1 in Leydig Cells Causes Male Reproductive Dysfunction in Mice." International Journal of Molecular Sciences 17.1 (2016): 96.  WB ;  Mouse.  
[IF=9.91] Li, Ting, et al. "Proliferation of parenchymal microglia is the main source of microgliosis after ischaemic stroke." Brain (2013): awt287.  Mouse.  
[IF=7.60] Teng, I., et al. "Phospholipid-functionalized mesoporous silica nanocarriers for selective photodynamic therapy of cancer." Biomaterials (2013).  WB ;  Mouse.  
[IF=4.75] Rosenzweig, Derek H., Sing J. Ou, and Thomas M. Quinn. ʺP38 mitogen‐activated protein kinase promotes dedifferentiation of primary articular chondrocytes in monolayer culture.ʺ Journal of Cellular and Molecular Medicine (2013)  WB ;  Bovine.  
[IF=4.26] Rosenzweig, Derek H., et al. "Mechanical injury of bovine cartilage explants induces depth-dependent, transient changes in MAP kinase activity associated with apoptosis." Osteoarthritis and Cartilage (2012).  WB ;  Bovine.  
[IF=4.22] Sondag, Gregory R., et al. "Osteoactivin Induces Transdifferentiation of C2C12 Myoblasts into Osteoblasts." Journal of Cellular Physiology (2013).  Mouse.  
[IF=3.95] Wang, Gang, et al. "Inhibition of hydrogen sulfide synthesis provides protection for severe acute pancreatitis rats via apoptosis pathway." Apoptosis (2013): 1-15.  IHC-P ;  Rat.  
[IF=3.06] Li, Guiying, et al. ʺGremlin Aggravates Hyperglycemia‐Induced Podocyte Injury by a TGFβ/Smad Dependent Signaling Pathway.ʺ Journal of CellularBiochemistry (2013).  WB ;  Mouse.  
[IF=1.83] KASSAYOVÁ, MONIKA, et al. "Anticancer and Immunomodulatory Effects of Lactobacillus plantarum LS/07, Inulin and Melatonin in NMU-induced Rat Model of Breast Cancer." Anticancer Research 36.6 (2016): 2719-2728.  IHC-P ;  Rat.  
[IF=1.72] Zhao, Jun, et al. "The cytotoxic and pro-apoptotic effects of phenylephrine on corneal stromal cells via a mitochondrion-dependent pathway both in vitro and in vivo." Experimental and Toxicologic Pathology (2016).  ELISA ;  Human.  
[IF=6.01] Chen, Zhoujiang, et al. "Tunable conjugation densities of camptothecin on hyaluronic acid for tumor targeting and reduction-triggered release." Acta Biomaterialia (2016).  IHC-P ;  Mouse.  
[IF=3.92] Wei, Gang, et al. "β-Asarone inhibits neuronal apoptosis via the CaMKII/CREB/Bcl-2 signaling pathway in an in vitro model and AβPP/PS1 mice." Journal of Alzheimer's Disease 33.3 (2013): 863-880.  IF(ICC) ;  Mouse.  
[IF=1.48] Liu, Yong‑Chao, et al. "RNAi-mediated gene silencing of vascular endothelial growth factor C suppresses growth and induces apoptosis in mouse breast cancer in vitro and in vivo." Oncology Letters. (2016)  WB ;  Mouse.  
[IF=2.61] Lei, Mingxing, et al. ʺGsdma3 is a new factor needed for TNF-α-mediated apoptosis signal pathway in mouse skin keratinocytes.ʺ Histochemistry and cell biology 138.3 (2012): 385-396.  WB, IHC-P ;  Mouse.  
[IF=2.41] Guan, Junhong, et al. "Bone morphogenetic protein-7 (BMP-7) mediates ischemic preconditioning-induced ischemic tolerance via attenuating apoptosis in rat brain." Biochemical and Biophysical Research Communications (2013).  WB ;  Rat.  
[IF=2.27] Li, M., et al. ʺmiR‐34c works downstream of p53 leading to dairy goat male germline stem‐cell (mGSCs) apoptosis.ʺ Cell Proliferation 46.2 (2013): 223-231.  WB ;  Goat.  
[IF=2.03] Aksu, Ilkay, et al. "Anxiety correlates to decreased blood and prefrontal cortex IGF-1 levels in streptozotocin induced diabetes." Neuroscience Letters (2012).  Rat.  
[IF=1.85] Li, Mingzhao, et al. ʺExpression of miR‐34c in response to overexpression of Boule and Stra8 in dairy goat male germ line stem cells (mGSCs).ʺ Cell Biochemistry and Function (2013).  Goat.  
[IF=2.13] Cetintas, Vildan Bozok, et al. "Effects of flavopiridol on critical regulation pathways of CD133high/CD44high lung cancer stem cells." Medicine 95.43 (2016): e5150.  IF(ICC) ;  Mouse.  
[IF=3.85] Wang, Yandi, et al. "Regulation of steroid hormones and energy status with cysteamine and its effect on spermatogenesis." Toxicology and Applied Pharmacology (2016).  IHC-P ;  Sheep.  
[IF=5.23] Zhao, Yong, et al. "Hydrogen Sulfide and/or Ammonia Reduces Spermatozoa Motility through AMPK/AKT Related Pathways." Scientific Reports 6 (2016): 37884.  WB ;  Pig.  
[IF=1.64] Kosutova, P., et al. "Intravenous dexamethasone attenuated inflammation and influenced apoptosis of lung cells in an experimental model of acute lung injury." Physiological research 65.Supplementum 5 (2016): S663.  IHC-P ;  Rabbit.  
[IF=2.38] Abdel-Hamid, Nagwa I., Mona F. El-Azab, and Yasser M. Moustafa. "Macrolide antibiotics differentially influence human HepG2 cytotoxicity and modulate intrinsic/extrinsic apoptotic pathways in rat hepatocellular carcinoma model." Naunyn-Schmiedeberg's Archives of Pharmacology (2017): 1-17.  IHC-P ;  Rat.  
[IF=1.55] Yang, Jinjiang, Ying Lu, and Ai Guo. "Platelet-rich plasma protects rat chondrocytes from interleukin-1β-induced apoptosis." Molecular Medicine Reports 14.5 (2016): 4075-4082.  WB ;  Rat.  
[IF=0.87] Ochigbo, Grace Onyeche, et al. "Polyphenol-rich fraction of Parquetina nigrescens mitigates dichlorvos-induced neurotoxicity and apoptosis." Journal of Ayurveda and Integrative Medicine (2017).  Rat.  
[IF=1.56] Qu, Zhongyuan, et al. "Chelidonine induces mitotic slippage and apoptotic-like death in SGC-7901 human gastric carcinoma cells." Molecular medicine reports 13.2 (2016): 1336-1344.  WB ;  Human.  
[IF=2.33] Yang, Juan, Guiyuan Li, and Keqiang Zhang. "Pro-survival effects by NF-κB, Akt and ERK (1/2) and anti-apoptosis actions by Six1 disrupt apoptotic functions of TRAIL-Dr4/5 pathway in ovarian cancer." Biomedicine & Pharmacotherapy 84 (2016): 1078-1087.  WB ;  Human.  
[IF=3.23] Wang, Yu, et al. "Ibutilide treatment protects against ER stress induced apoptosis by regulating calumenin expression in tunicamycin treated cardiomyocytes." PloS one 12.4 (2017): e0173469.  WB ;  Rat.  
[IF=4.94] Kubatka, Peter, et al. "Antineoplastic effects of clove buds (Syzygium aromaticum L.) in the model of breast carcinoma." Journal of Cellular and Molecular Medicine (2017).  IHC-P ;  Rat.  
[IF=5.47] Pan, Bo, et al. "c-Abl Tyrosine Kinase Mediates Neurotoxic Prion Peptide-Induced Neuronal Apoptosis via Regulating Mitochondrial Homeostasis." Molecular Neurobiology (2014): 1-15.  Rat.  
[IF=2.75] Ye, Bengui, et al. "Anti-tumor activity and relative mechanism of ethanolic extract of Marsdenia tenacissima (Asclepiadaceae) against human hematologic neoplasm in vitro and in vivo." Journal of Ethnopharmacology (2014).  WB ;  Human.  
[IF=2.69] Lin, Caiyu, et al. "Lithocarpus Polystachyus Rehd Leaf Aqueous Extract Inhibits Human Breast Cancer Growth In Vitro and In Vivo." Nutrition and Cancer (2014).  WB ;  Human.  
[IF=3.17] Liu, Yang, et al. "Amelioration of Stroke-Induced Neurological Deficiency by Lyophilized Powder of Catapol and Puerarin." International Journal of Biological Sciences 10.4 (2014): 448-456.  WB ;  Mouse.  
[IF=2.52] Yao, Kun, et al. "Knockdown of RLIP76 expression by RNA interference inhibits proliferation, enhances apoptosis, and increases chemosensitivity to daunorubicin in U937 leukemia cells." Tumor Biology (2014): 1-9.  WB ;  Human.  
[IF=2.15] Tang, Xi-Lan, et al. "The Cardioprotective Effect of Protocatechuic Acid on Myocardial Ischemia/Reperfusion Injury." Journal of Pharmacological Sciences 125.2 (2014): 176-183.  WB ;  Rat.  
[IF=3.13] Zhang, Wen-feng, et al. "Angelica polysaccharides inhibit the growth and promote the apoptosis of U251 glioma cells in vitro and in vivo." Phytomedicine (2017).  WB ;  Human.  
[IF=2.10] Zhang, Beiru, et al. "The Toxicity Mechanisms of Action of Aβ25–35 in Isolated Rat Cardiac Myocytes." Molecules 19.8 (2014): 12242-12257.  other ;  Rat.  
[IF=3.79] Luo, Xiaoming, et al. "Antimetastasis and antitumor efficacy promoted by sequential release of vascular disrupting and chemotherapeutic agents from electrospun fibers." International Journal of Pharmaceutics (2014).  IHC-P ;  Mouse.  
[IF=2.38] Gao, Hui, et al. "Hispidulin Potentiates the Antitumor Effect of Sunitinib Against Human Renal Cell Carcinoma in Laboratory Models." Cell Biochemistry and Biophysics: 1-8.  WB ;  Human.  
[IF=2.38] Gao, Hui, et al. "shRNA-Mediated EMMPRIN Silencing Inhibits Human Leukemic Monocyte Lymphoma U937 Cell Proliferation and Increases Chemosensitivity to Adriamycin." Cell Biochemistry and Biophysics: 1-9.  WB ;  Human.  
[IF=3.68] Xiao, Ke, et al. "The role of visfatin on the regulation of inflammation and apoptosis in the spleen of LPS-treated rats." Cell and Tissue Research (2014).  IHC-P ;  Rat.  
[IF=3.53] Fang C, Zhang J, Qi D, Fan X, Luo J, et al. (2014) Evodiamine Induces G2/M Arrest and Apoptosis via Mitochondrial and Endoplasmic Reticulum Pathways in H446 and H1688 Human Small-Cell Lung Cancer Cells. PLoS ONE 9(12): e115204.   WB ;  Human.  
[IF=1.14] Wang, J., et al. "Esculetin, a coumarin derivative, exerts in vitro and in vivo antiproliferative activity against hepatocellular carcinoma by initiating a mitochondrial-dependent apoptosis pathway." Brazilian Journal of Medical and Biological Research (2014): 000-000.  WB ;  Mouse.  
[IF=4.20] Tian, A., et al. "Nanoscale TiO 2 nanotubes govern the biological behavior of human glioma and osteosarcoma cells." International Journal of Nanomedicine10 (2015): 2377-2389.  Human.  
[IF=3.84] Kubatka, Peter, et al. "Fruit peel polyphenols demonstrate substantial anti-tumour effects in the model of breast cancer." European Journal of Nutrition(2015): 1-11.  IHC-P ;  Rat.  
[IF=1.55] Jia, Jianping, et al. "A preliminary study of the effect of ECRG4 overexpression on the proliferation and apoptosis of human laryngeal cancer cells and the underlying mechanisms." Molecular Medicine Reports.  Human.  
[IF=2.37] Liu, Jun-Song, et al. "β-elemene enhances the radiosensitivity of gastric cancer cells by inhibiting Pak1 activation." World J Gastroenterol 21.34 (2015): 9945-9956.  WB ;  Human.  
[IF=1.55] Song, Guijun, et al. "Anti-autophagic and anti-apoptotic effects of memantine in a SH-SY5Y cell model of Alzheimers disease via mammalian target of rapamycin-dependent and-independent pathways." Molecular Medicine Reports.  WB ;  Human.  
[IF=2.70] Lv, Runxiao, et al. "Neuroprotective effect of allicin in a rat model of acute spinal cord injury." Life Sciences (2015).  WB ;  Rat.  
[IF=2.31] Dong, Liwei, et al. "Echinacoside induces apoptotic cancer cell death by inhibiting the nucleotide pool sanitizing enzyme MTH1." OncoTargets and Therapy 8 (2015): 3649.  Human.  
[IF=0.00] Wang, Yu, et al. "Tanshinone II A Relieves Adriamycin-induced Myocardial Injury in Rat Model." International Journal of Chemistry 8.1 (2016): 40.  WB ;  Rat.  
[IF=3.82] Clark, Andrea J., et al. "WO3/Pt nanoparticles are NADPH oxidase biomimetics that mimic effector cells in vitro and in vivo." Nanotechnology27.6 (2015): 065101.  
规格价格 50ul/780元 购买    100ul/1380元 购买    200ul/2200元 购买    大包装/询价
说 明 书 50ul  100ul  200ul
研究领域 肿瘤  细胞生物  神经生物学  信号转导  细胞凋亡  Alzheimer's  
抗体来源 Rabbit
克隆类型 Polyclonal
交叉反应 Human, Mouse, Rat, Dog, Pig, Cow, Horse, Rabbit, Sheep, 
产品应用 WB=1:500-2000 ELISA=1:500-1000 IHC-P=1:400-800 IHC-F=1:400-800 Flow-Cyt=2μg/Test IF=1:100-500 (石蜡切片需做抗原修复)
not yet tested in other applications.
optimal dilutions/concentrations should be determined by the end user.
分 子 量 28kDa
细胞定位 细胞浆 
性    状 Lyophilized or Liquid
浓    度 1mg/ml
免 疫 原 KLH conjugated synthetic peptide derived from human caspase-3 p17 subunit:1-100/277 
亚    型 IgG
纯化方法 affinity purified by Protein A
储 存 液 0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol.
保存条件 Store at -20 °C for one year. Avoid repeated freeze/thaw cycles. The lyophilized antibody is stable at room temperature for at least one month and for greater than a year when kept at -20°C. When reconstituted in sterile pH 7.4 0.01M PBS or diluent of antibody the antibody is stable for at least two weeks at 2-4 °C.
PubMed PubMed
产品介绍 background:
The caspase family of cysteine proteases play a key role in apoptosis. Caspase 3 is the most extensively studied apoptotic protein among caspase family members. Caspase 3 is synthesized as inactive pro enzyme that is processed in cells undergoing apoptosis by self proteolysis and/or cleavage by other upstream proteases (e.g. Caspases 8, 9 and 10). The processed form of Caspase 3 consists of large (17kDa) and small (12kDa) subunits which associate to form an active enzyme. Caspase 3 is cleaved at Asp28 Ser29 and Asp175 Ser176. The active Caspase 3 proteolytically cleaves and activates other caspases (e.g. Caspases 6, 7 and 9), as well as relevant targets in the cells (e.g. PARP and DFF). Alternative splicing of this gene results in two transcript variants which encode the same protein. In immunohistochemical studies Caspase 3 expression has been shown to be widespread but not present in all cell types (e.g. commonly reported in epithelial cells of skin, renal proximal tubules and collecting ducts). Differences in the level of Caspase 3 have been reported in cells of short lived nature (eg germinal centre B cells) and those that are long lived (eg mantle zone B cells). Caspase 3 is the predominant caspase involved in the cleavage of amyloid beta 4A precursor protein, which is associated with neuronal death in Alzheimer's disease.
Reacts with Caspase-3 subunit p17 and precursor.

Function:
Involved in the activation cascade of caspases responsible for apoptosis execution. At the onset of apoptosis it proteolytically cleaves poly(ADP-ribose) polymerase (PARP) at a '216-Asp-|-Gly-217' bond. Cleaves and activates sterol regulatory element binding proteins (SREBPs) between the basic helix-loop-helix leucine zipper domain and the membrane attachment domain. Cleaves and activates caspase-6, -7 and -9. Involved in the cleavage of huntingtin. Triggers cell adhesion in sympathetic neurons through RET cleavage.

Subunit:
Heterotetramer that consists of two anti-parallel arranged heterodimers, each one formed by a 17 kDa (p17) and a 12 kDa (p12) subunit. Interacts with BIRC6/bruce.

Subcellular Location:
Cytoplasm.

Tissue Specificity:
Highly expressed in lung, spleen, heart, liver and kidney. Moderate levels in brain and skeletal muscle, and low in testis. Also found in many cell lines, highest expression in cells of the immune system.

Post-translational modifications:
Cleavage by granzyme B, caspase-6, caspase-8 and caspase-10 generates the two active subunits. Additional processing of the propeptides is likely due to the autocatalytic activity of the activated protease. Active heterodimers between the small subunit of caspase-7 protease and the large subunit of caspase-3 also occur and vice versa.
S-nitrosylated on its catalytic site cysteine in unstimulated human cell lines and denitrosylated upon activation of the Fas apoptotic pathway, associated with an increase in intracellular caspase activity. Fas therefore activates caspase-3 not only by inducing the cleavage of the caspase zymogen to its active subunits, but also by stimulating the denitrosylation of its active site thiol.

Similarity:
Belongs to the peptidase C14A family.

SWISS:
P42574

Gene ID:
836

Database links:

Entrez Gene: 836 Human

Entrez Gene: 12367 Mouse

Entrez Gene: 397244 Pig

Entrez Gene: 100008840 Rabbit

Entrez Gene: 25402 Rat

Omim: 600636 Human

SwissProt: P42574 Human

SwissProt: P70677 Mouse

SwissProt: Q95ND5 Pig

SwissProt: Q8MJC3 Rabbit

SwissProt: P55213 Rat

Unigene: 141125 Human

Unigene: 34405 Mouse

Unigene: 10562 Rat



Important Note:
This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.

Caspase3广泛分布于各种不同类型的细胞中,是Caspase家族中最重要的凋亡执行者之一,激活的Caspase-3能使许多与细胞结构、细胞周期及DNA修复等相关蛋白或激酶失活,从而使细胞凋亡.
产品图片
Protein:Lung(Mouse) lysate at 30ug;
Primary: Anti-caspase-3 (bs-0081R) at 1:300 dilution;
Secondary: HRP conjugated Goat-Anti-Rabbit IgG(bse-0295G) at 1: 5000;
Predicted band size : 17/28 kD
Observed band size : 28 kD
Sample:
Kidney (Mouse) Lysate at 40 ug
Primary: Anti-Caspase-3 (bs-0081R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 28 kD
Observed band size: 17 kD
Sample:
Liver (Mouse) Lysate at 40 ug
Primary: Anti-Caspase-3 (bs-0081R) at 1/500 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 28 kD
Observed band size: 28 kD
Sample: Raji Cell lysate;
Primary: Anti-Caspase-3 (bs-0081R) at 1:300;
Secondary: HRP conjugated Goat-Anti-rabbit IgG(bs-0295G-HRP) at 1: 5000;
Predicted band size: 17/28 kD
Observed band size: 26 kD
Sample:Liver(Sheep)Lysate at 40 ug
Primary: Anti-Caspase-3(bs-0081R)at 1/300 dilution
Secondary: IRDye800CW Goat Anti-RabbitIgG at 1/20000 dilution
Predicted band size: 28kD
Observed band size: 30kD
Tissue/cell: rat brain tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-Caspase-3 Polyclonal Antibody, Unconjugated(bs-0081R) 1:200, overnight at 4癈, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
Tissue/cell: rabbit pancreas tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-Caspase-3 Polyclonal Antibody, Unconjugated(bs-0081R) 1:300, overnight at 4癈, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
The figure annotation:
The blue histogram is unstained cells
. The Orange histogram is cells stained with Rabbit IgG/FITC (bs-0295P-FITC).
The green histogram is cells stained with Rabbit Anti-Caspase-3/FITC Conjugated antibody (bs-0081R-FITC).
Controls
Positive control: HepG 2 cells
Isotype control: Cell lines treated with Rabbit IgG/FITC (bs-0295P-FITC) instead of the primary antibody to confirm that primary antibody binding is specific. 2μg in 1 00μL1 X PBS containing 0.5% BSA.
Blank control: Mouse spleen.
Primary Antibody (green line): Rabbit Anti-Caspase-3 antibody (bs-0081R)
Dilution: 1μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Dilution: 1μg /test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at-20℃. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. Acquisition of 20,000 events was performed.
The figure annotation:
The blue histogram is unstained cells.
The Orange histogram is cells stained with Rabbit IgG/FITC (bs-0295P-FITC).
The green histogram is cells stained with Rabbit Anti-Caspase-3/FITC Conjugated antibody (bs-0081R-FITC).
Controls
Positive control: A549 cells
Isotype control: Cell lines treated with Rabbit IgG/FITC(bs-0295P-FITC) instead of the primary antibody to confirm that primary antibody binding is specific. 3μg in 1 00 μL 1 X PBS containing 0.5% BSA.
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