来源
用途
交叉反应性
理论分子量
实际分子量
Rabbit
WB, IF, IHC
H, M, R, Pg
14kDa
14/16kDa
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
1:50-1:200
-
-
-
抗体详细信息如下::
About this Antibody
Name
LC3B Rabbit Polyclonal Antibody (KO Validated)
Category
Rabbit Polyclonal Antibody (pAb); Primary antibody
Isotype
IgG
Purification method
Affinity purification
Positive samples
C6, HeLa, NIH/3T3, Pig liver
Cellular location
Cytoplasm, Cytoplasmic vesicle, Endomembrane system, Lipid-anchor, autophagosome, autophagosome membrane, cytoskeleton
Customer validation
WB (Mouse); IHC (Mouse)
About the Immunogen
Immunogen
A synthetic peptide of human LC3B
Sequence
-
Gene ID
81631
Swiss Prot
Q9GZQ8
Synonyms
MAP1LC3B; ATG8F; LC3B; MAP1A/1BLC3; MAP1LC3B-a; microtubule associated protein 1 light chain 3 beta
Category
Organelle Markers; Autophagy Signaling
Background
Autophagy is a catabolic process for the autophagosomic-lysosomal degradation of bulk cytoplasmic contents. Autophagy is generally activated by conditions of nutrient deprivation, but it has also been associated with a number of physiological processes including development, differentiation, neurodegenerative diseases, infection, and cancer. Autophagy marker Light Chain 3 (LC3) was originally identified as a subunit of microtubule-associated proteins 1A and 1B (termed MAP1LC3) and subsequently found to contain similarity to the yeast protein Apg8/Aut7/Cvt5 critical for autophagy. Three human LC3 isoforms (LC3A, LC3B, and LC3C) undergo post-translational modifications during autophagy. Cleavage of LC3 at the carboxy terminus immediately following synthesis yields the cytosolic LC3-I form. During autophagy, LC3-I is converted to LC3-II through lipidation by a ubiquitin-like system involving Atg7 and Atg3 that allows for LC3 to become associated with autophagic vesicles. The presence of LC3 in autophagosomes and the conversion of LC3 to the lower migrating form, LC3-II, have been used as indicators of autophagy.
包装清单：
产品编号
产品名称
包装
AF5225
LC3B Rabbit Polyclonal Antibody (KO Validated)
50μl
AZ050
Western一抗稀释液
50ml
—
说明书
1份
保存条件：
LC3B 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 various cell lines, using LC3B antibody at 1:1000 dilution. C6 cells were treated by Chloroquine for 20 hours. Hela cells were treated by Chloroquine for 20 hours. NIH/3T3 cells were treated by Chloroquine for 20 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 of pig liver, using LC3B antibody at 1:10000 dilution. Lysates/proteins: 25µg per lane. Blocking buffer: QuickBlock™ Blocking Buffer (P0231).
Fig. 3. Immunohistochemistry of paraffin-embedded rat heart using LC3B antibody at dilution of 1:100 (40x lens).
Fig. 4. Immunofluorescence analysis of NIH/3T3 cells using LC3B antibody at dilution of 1:100. NIH/3T3 cells were treated by Chloroquine (50 μM) for 20 hours(left). Blue: DAPI for nuclear staining.
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