来源
用途
交叉反应性
理论分子量
实际分子量
Rabbit
WB, IHC
H, M, R
31kDa
17/35kDa
WB, Western blot; IP, Immunoprecipitation; IF, Immunofluorescence; IHC, Immunohistochemistry; ICC, Immunocytochemistry; FC, Flow Cytometry; ChIP, Chromatin Immunoprecipitation Assay; ChIP-seq, ChIP-sequencing.
H, Human; M, Mouse; R, Rat; C, Chicken; Cw, Cow; Dg, Dog; Gp, Guinea pig; Hm, Hamster; Hr, Horse; Mk, Monkey; Pg, Pig; Rb, Rabbit; S, Sheep; Z, Zebrafish; All, all species expected.
配套提供了Western一抗稀释液，可以用于Western检测或其它适当用途时的一抗稀释。
建议抗体使用时的稀释比例如下(实际使用时需根据抗原水平的高低作适当调整)：
WB
IP
IF
IHC
ICC
FC
ChIP
ChIP-seq
1:500-1:2000
-
-
1:50-1:200
1:50-1:200
-
-
-
抗体详细信息如下::
About this Antibody
Name
Caspase 3 Rabbit Polyclonal Antibody (KO Validated)
Category
Rabbit Polyclonal Antibody (pAb); Primary antibody
Isotype
IgG
Purification method
Affinity purification
Positive samples
Jurkat
Cellular location
Cytoplasm
Customer validation
WB (Human, P. notoginseng, Mouse, Rat, Pig, Chicken)
About the Immunogen
Immunogen
Recombinant fusion protein of human Caspase 3 (NP_004337.2).
Sequence
FHKSTGMTSRSGTDVDAANLRETFRNLKYEVRNKNDLTREEIVELMRDVSKEDHSKRSSFVCVLLSHGEEGIIFGTNGPVDLKKITNFFRGDRCRSLTGKPKLFII
Gene ID
836
Swiss Prot
P42574
Synonyms
CASP3; CPP32; CPP32B; SCA-1; caspase-3
Category
Regulation of Apoptosis; Mitochondrial Control of Apoptosis; Amyloid Plaque and Neurofibrillary Tangle Formation in Alzheimer's Disease; Death Receptor Signaling
Background
Caspase-3 (Cysteine-aspartic acid protease 3/Casp3; also Yama, apopain and CPP32) is a 29 kDa member of the peptidase C14A family of enzymes. It is widely expressed and is an integral component of the apoptotic cascade. Caspase-3 is considered to be the major executioner caspase; that is, the primary downstream mediator of apoptotic-associated proteolysis. Active Caspase-3 is known to utilize a Cys residue to cleave multiple substrates, including PARP, proIL 16, PKC-gamma & -δ, procaspases 6, 7 and 9, and beta catenin. Human procaspase-3 is a 32 kDa, 277 amino acid (aa) protein. Normally, it is an inactive, cytosolic homodimer, but following an upstream signal that activates processing proteases, procaspase-3 undergoes proteolytic cleavage. This generates an N-terminal 175 aa p20/20 kDa subunit plus a 102 aa C-terminal p12/12 kDa subunit, followed by further processing of the p20 subunit at Asp28 to generate a final p17 subunit (aa 29 175) The p17 and p12 subunits noncovalently heterodimerize, and subsequently associate with another p17/p12 heterodimer to form an active antiparallel homodimer. The p17 subunit contains the enzyme active site (aa 161 165), with an embedded catalytic Cys which is normally nitrosylated and inactive. Full activation requires both proteolytic processing and Cys163 denitrosylation. Multiple proteases can use Caspase-3 as a substrate including Caspase-6, -8, and -10, granzyme B, and Caspase-3 itself.
包装清单：
产品编号
产品名称
包装
AF5132
Caspase 3 Rabbit Polyclonal Antibody (KO Validated)
50μl
AZ050
Western一抗稀释液
50ml
—
说明书
1份
保存条件：
Caspase 3 Rabbit Polyclonal Antibody (KO Validated) -20ºC保存，Western一抗稀释液-20ºC或4ºC保存，一年有效。Western一抗稀释液优先推荐4ºC保存，长期不使用可以考虑-20ºC保存，但冻融可能会导致出现轻微的浑浊和少量不溶物。
注意事项：
如果本抗体用于Western blot (WB)、免疫荧光(IF)、免疫细胞化学(ICC)等实验，请注意回收使用过的稀释抗体。回收的抗体通常至少可以重复使用5-10次。稀释后的抗体，包括已经使用过的稀释抗体，请4℃保存。
回收后重复使用的抗体，使用方法同新鲜稀释的抗体。如果在重复使用过程中发现抗体出现轻微混浊现象，可以10,000g离心1-3分钟，取上清用于后续检测。如果回收的抗体出现明显的絮状物或长霉长菌等情况，则可以考虑废弃该抗体。
提供的Western一抗稀释液也可以用于免疫荧光(IF)、免疫组化(IHC)、免疫细胞化学(ICC)等适当用途。如果希望获得最佳的检测效果，请考虑使用上述检测专用的一抗稀释液。
本产品仅限于专业人员的科学研究用，不得用于临床诊断或治疗，不得用于食品或药品，不得存放于普通住宅内。
为了您的安全和健康，请穿实验服并戴一次性手套操作。
使用说明：
请根据抗体的实际用途选择相应的使用方法。
1. Western检测：
a. 按照推荐的稀释比例用提供的Western一抗稀释液稀释抗体。
b. 把经过封闭的蛋白膜与稀释好的一抗4℃缓慢摇动过夜或室温缓慢摇动2小时，确保稀释的抗体至少能在摇动的瞬间覆盖蛋白膜。
c. 回收稀释的一抗，4℃保存以备下次继续使用。
d. 按照Western的实验步骤进行后续的洗涤、二抗孵育、洗涤和检测等操作。
2. 免疫染色：
可以使用生产的免疫染色一抗稀释液(P0103)稀释抗体，使用后注意回收稀释好的一抗，
3. 其它实验操作请自行参考适当的protocol进行。
4. 代表性图片：
Fig. 1. Western blot analysis of extracts of Jurkat cells, using Caspase 3 antibody at 1:1000 dilution. Jurkat cells treated by Etoposide 25uM etoposide for 5 hours. Secondary antibody: HRP-labeled Goat Anti-Rabbit IgG(H+L) (A0208) at 1:1000 dilution. Lysates/proteins: 25µg per lane. Blocking buffer: QuickBlock™ Blocking Buffer (P0231). Detection: BeyoECL Star (P0018A). Exposure time:1s.
Fig. 2. Western blot analysis of extracts from normal (control) and Caspase 3 knockout (KO) 293T cells, using Caspase 3 antibody at 1:1000 dilution. Secondary antibody: HRP-labeled Goat Anti-Rabbit IgG(H+L) (A0208) at 1:1000 dilution. Lysates/proteins: 25µg per lane. Blocking buffer: QuickBlock™ Blocking Buffer (P0231). Detection: BeyoECL Star (P0018A). Exposure time:1s.
Fig. 3. Immunohistochemistry of paraffin-embedded rat spleen using Caspase 3 antibody at dilution of 1:100 (40x lens).
Fig. 4. Immunohistochemistry of paraffin-embedded human tonsil using Caspase 3 antibody at dilution of 1:100 (40x lens).
​