Cell Signaling Technology

Product Pathways - Autophagy Signaling

LC3B (D11) XP® Rabbit mAb #3868

lc3   lc3-b   sc-16756   sc-28266  

No. Size Price
3868S 100 µl ( 10 western blots ) ¥3,580.00 现货查询 购买询价
3868T 20 µl ( 2 western blots ) ¥1,400.00 现货查询 购买询价
3868 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat, Endogenous 14, 16 Rabbit IgG
IP 1:50
IHC-P 1:3200
F 1:400
IF-IC 1:200

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, IHC-P=Immunohistochemistry (Paraffin), F=Flow Cytometry, IF-IC=Immunofluorescence (Immunocytochemistry),

Homology

Species predicted to react based on 100% sequence homology: Monkey, Bovine, Pig,

Specificity / Sensitivity

LC3B (D11) XP® Rabbit mAb detects endogenous levels of total LC3B protein. Cross-reactivity may occur with other LC3 isoforms. Stronger reactivity is observed with the type II form of LC3B.

LC3B (D11) XP® Rabbit mAb 兔单抗能检测内源性的LC3B总蛋白。可能会与其他LC3亚型发生交叉反应。与LC3B的II型形式有较强的反应性。

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of LC3B.

该单克隆抗体是用合成的LC3B氨基末端附近的残基相对应的肽段免疫动物制备。

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells, untreated (left) or chloroquine-treated (right), using LC3B (D11) XP® Rabbit mAb (green). Actin filaments were labeled using DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

共聚焦免疫荧光分析HeLa细胞,未处理(左)或氯喹处理(右),使用的抗体是LC3B (D11) XP® Rabbit mAb 兔单抗(绿色)。肌动蛋白丝用DY-554鬼笔环肽标记(红色)。蓝色伪彩= DRAQ5® #4084(DNA荧光染料)。

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of HeLa cells using LC3B (D11) XP® Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).

流式细胞仪分析HeLa细胞,使用的抗体是LC3B (D11) XP® Rabbit mAb 兔单抗(蓝色),一种非特异的阴性对照抗体(红色)作为对照。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded HeLa cell pellets, control (left) or chloroquine-treated (right), using LC3B (D11) XP® Rabbit mAb.

免疫组化分析石蜡包埋的HeLa细胞沉淀,对照(左)或 chloroquine (氯喹)处理(右),使用的抗体是LC3B (D11) XP® Rabbit mAb 兔单抗。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human astrocytoma using LC3B (D11) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).

免疫组化分析石蜡包埋的人星形细胞瘤,使用的抗体是LC3B (D11) XP® Rabbit mAb 兔单抗,分别经对照肽(左)或 抗原特异性多肽 (右)处理。

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® LC3B siRNA I #6212 (+) or SignalSilence® LC3B siRNA II #6213 (+), using LC3B (D11) XP® Rabbit mAb #3868 and α-Tubulin (11H10) Rabbit mAb #2125. The LC3B (D11) XP® Rabbit mAb confirms silencing of LC3B expression, while the α-Tubulin (11H10) Rabbit mAb is used to control for loading and specificity of LC3B siRNA.

Western blot分析HeLa细胞的提取物,分别用100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-),SignalSilence® LC3B siRNA I #6212 (+)或SignalSilence® LC3B siRNA II #6213 (+)转染。使用的抗体是:LC3B (D11) XP® Rabbit mAb 兔单抗#3868 和 α-Tubulin (11H10) Rabbit mAb 兔单抗#2125。α-Tubulin (11H10) Rabbit mAb 兔单抗作为加样和LC3B siRNA的特异性对照,LC3B (D11) XP® Rabbit mAb 兔单抗证实LC3B 表达的沉默。

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines, untreated (-) or treated overnight with chloroquine (50 μM) (+), using LC3B (D11) XP® Rabbit mAb (upper) or LC3B Antibody #2775 (lower).

Western blot分析不同细胞系的提取物,未处理(-)或chloroquine(氯喹)处理(50 μM,过夜,+),使用的抗体分别是LC3B (D11) XP® Rabbit mAb 兔单抗(上图)和LC3B Antibody #2775(下图)。

IF-IC

IF-IC

Confocal immunofluorescent analysis of HCT-116 cells, untreated (left) or choroquine-treated (50 uM, overnight; right) using LC3B (D11) XP® Rabbit mAb (green) and β-Catenin (L54E2) Mouse mAb (Alexa Fluor® 555 Conjugate) #5612 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Background

Autophagy is a catabolic process for the autophagosomic-lysosomal degradation of bulk cytoplasmic contents (1,2). Autophagy is generally activated by conditions of nutrient deprivation but has also been associated with a number of physiological processes including development, differentiation, neurodegenerative diseases, infection and cancer (3). Autophagy marker Light Chain 3 (LC3) was originally identified as a subunit of microtubule-associated proteins 1A and 1B (termed MAP1LC3) (4), and subsequently found to contain similarity to the yeast protein Apg8/Aut7/Cvt5 critical for autophagy (5). Three human LC3 isoforms (LC3A, LC3B, and LC3C) undergo post-translational modifications during autophagy (6-9). 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 (6-10). 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 (11).

自噬是自噬溶酶体对其包裹的细胞质内含物进行降解的一种分解代谢过程(1,2)。自噬过程一般是在营养匮乏的条件下被激活,但是也与一些生理过程有关,如发育、分化、神经变性、感染和癌症(3)。自噬微管相关蛋白的轻链3(LC3)最初被认定为是微管相关蛋白1A和1B的一个亚基(被称为MAP1LC3)(4),随后发现该蛋白所包含的与酵母蛋白类似的Apg8/Aut7/Cvt5对于自噬起着至关重要的作用(5)。三种人类LC3亚型(LC3A、LC3B和LC3C)在自噬过程中翻译后被修饰(6-9)。随着合成后LC3羧基末端迅速裂解,产生了胞浆的LC3-I 型。在自噬过程中,通过一个涉及Atg7和Agt3的类泛素体系,LC3-I被脂质化为LC3-II,从而将LC3与自噬小泡联系起来(6-10)。自噬体中LC3的存在,及其向低迁移形式的LC3-II的转化被作为自噬发生的“指示器”(11)。

  1. Reggiori, F. and Klionsky, D.J. (2002) Eukaryot. Cell 1, 11-21.
  2. Codogno, P. and Meijer, A.J. (2005) Cell Death Differ. 12 Suppl 2, 1509-18.
  3. Levine, B. and Yuan, J. (2005) J. Clin. Invest. 115, 2679-88.
  4. Mann, S.S. and Hammarback, J.A. (1994) J. Biol. Chem. 269, 11492-97.
  5. Lang, T. et al. (1998) EMBO J. 17, 3597-607.
  6. Kabeya, Y. et al. (2000) EMBO J. 19, 5720-28.
  7. He, H. et al. (2003) J. Biol. Chem. 278, 29278-87.
  8. Tanida, I. et al. (2004) J. Biol. Chem. 279, 47704-10.
  9. Wu, J. et al. (2006) Biochem. Biophys. Res. Commun. 339, 437-42.
  10. Ichimura, Y. et al. (2000) Nature 408, 488-92.
  11. Kabeya, Y. et al. (2004) J. Cell Sci. 117, 2805-12.

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Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

XP is a registered trademark of Cell Signaling Technology, Inc.

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