LC3A/B Antibody - #AF5402

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产品: LC3A/B 抗体
货号: AF5402
描述: Rabbit polyclonal antibody to LC3A/B
应用: WB IHC IF/ICC
反应: Human, Mouse, Rat
预测: Pig, Zebrafish, Bovine, Sheep, Dog, Xenopus
分子量: 14kDa,16 kDa; 14kD,15kD(Calculated).
蛋白号: Q9H492 | Q9GZQ8
RRID: AB_2837886

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说明书
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实验方案
WB Handbook
IHC Protocol
IF/ICC Protocol

产品描述

来源:
Rabbit
应用:
WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: 适用于变性蛋白样本的免疫印迹检测. IHC: 适用于组织样本的石蜡(IHC-p)或冰冻(IHC-f)切片样本的免疫组化/荧光检测. IF/ICC: 适用于细胞样本的荧光检测. ELISA(peptide): 适用于抗原肽的ELISA检测.

反应:
Human,Mouse,Rat
预测:
Pig(100%), Zebrafish(93%), Bovine(100%), Sheep(100%), Dog(100%), Xenopus(100%)
克隆:
Polyclonal
特异性:
LC3A/B Antibody detects endogenous levels of total LC3A/B.
RRID:
AB_2837886
引用格式: Affinity Biosciences Cat# AF5402, RRID:AB_2837886.
偶联:
Unconjugated.
纯化:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
保存:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
别名:

展开/折叠

ATG8E; Autophagy-related protein LC3 A; Autophagy-related ubiquitin-like modifier LC3 A; LC3; LC3A; MAP1 light chain 3 like protein 1; MAP1 light chain 3-like protein 1; MAP1A/1B light chain 3 A; MAP1A/MAP1B LC3 A; MAP1A/MAP1B light chain 3 A; MAP1ALC3; MAP1BLC3; Map1lc3a; Microtubule associated proteins 1A/1B light chain 3; Microtubule-associated protein 1 light chain 3 alpha; Microtubule-associated proteins 1A and 1B, light chain 3; Microtubule-associated proteins 1A/1B light chain 3A; MLP3A_HUMAN; ATG8F; Autophagy-related protein LC3 B; Autophagy-related ubiquitin-like modifier LC3 B; LC3B; LC3II; MAP1 light chain 3 like protein 2; MAP1 light chain 3-like protein 2; MAP1A/1BLC3; MAP1A/MAP1B LC3 B; MAP1A/MAP1B light chain 3 B; MAP1ALC3; MAP1LC3B a; Map1lc3b; Microtubule associated protein 1 light chain 3 beta; Microtubule-associated protein 1 light chain 3 beta; Microtubule-associated proteins 1A/1B light chain 3B; MLP3B_HUMAN;

抗原和靶标

免疫原:
Uniprot:

Q9H492(MLP3A_HUMAN) >>Visit HPA database.

Q9GZQ8(MLP3B_HUMAN) >>Visit HPA database.

基因/基因ID:
MAP1LC3A(84557) | MAP1LC3B(81631)
表达:
Q9H492 MLP3A_HUMAN:

Most abundant in heart, brain, liver, skeletal muscle and testis but absent in thymus and peripheral blood leukocytes.

Q9GZQ8 MLP3B_HUMAN:

Most abundant in heart, brain, skeletal muscle and testis. Little expression observed in liver.

描述:
Ubiquitin-like modifier involved in formation of autophagosomal vacuoles (autophagosomes). Plays a role in mitophagy which contributes to regulate mitochondrial quantity and quality by eliminating the mitochondria to a basal level to fulfill cellular energy requirements and preventing excess ROS production.
序列:
MPSDRPFKQRRSFADRCKEVQQIRDQHPSKIPVIIERYKGEKQLPVLDKTKFLVPDHVNMSELVKIIRRRLQLNPTQAFFLLVNQHSMVSVSTPIADIYEQEKDEDGFLYMVYASQETFGF

MPSEKTFKQRRTFEQRVEDVRLIREQHPTKIPVIIERYKGEKQLPVLDKTKFLVPDHVNMSELIKIIRRRLQLNANQAFFLLVNGHSMVSVSTPISEVYESEKDEDGFLYMVYASQETFGMKLSV

种属预测

种属预测:

score>80的预测可信度较高，可尝试用于WB检测。*预测模型主要基于免疫原序列比对，结果仅作参考，不作为质保凭据。

Species
Results
Score
Pig
100
Bovine
100
Sheep
100
Dog
100
Xenopus
100
Zebrafish
93
Horse
0
Chicken
0
Rabbit
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

翻译修饰 - Q9H492/Q9GZQ8 作为底物

Site PTM Type Enzyme
S12 Phosphorylation P17612 (PRKACA)
K42 Ubiquitination
K49 Acetylation
K49 Ubiquitination
T50 Phosphorylation Q13188 (STK3) , Q13043 (STK4)
K51 Acetylation
S92 Phosphorylation
T93 Phosphorylation
Site PTM Type Enzyme
K5 Ubiquitination
T6 Phosphorylation
R21 Methylation
T29 Phosphorylation
K30 Ubiquitination
K42 Ubiquitination
K49 Ubiquitination
K51 Ubiquitination
K65 Ubiquitination

研究背景

功能:

Ubiquitin-like modifier involved in formation of autophagosomal vacuoles (autophagosomes). Whereas LC3s are involved in elongation of the phagophore membrane, the GABARAP/GATE-16 subfamily is essential for a later stage in autophagosome maturation. Through its interaction with the reticulophagy receptor TEX264, paticipates in the remodeling of subdomains of the endoplasmic reticulum into autophagosomes upon nutrient stress, which then fuse with lysosomes for endoplasmic reticulum turnover.

翻译修饰:

The precursor molecule is cleaved by ATG4B to form the cytosolic form, LC3-I. This is activated by APG7L/ATG7, transferred to ATG3 and conjugated to phospholipid to form the membrane-bound form, LC3-II.

The Legionella effector RavZ is a deconjugating enzyme that produces an ATG8 product that would be resistant to reconjugation by the host machinery due to the cleavage of the reactive C-terminal glycine.

Phosphorylation at Ser-12 by PKA inhibits conjugation to phosphatidylethanolamine (PE). Interaction with MAPK15 reduces the inhibitory phosphorylation and increases autophagy activity.

细胞定位:

Cytoplasm>Cytoskeleton. Endomembrane system>Lipid-anchor. Cytoplasmic vesicle>Autophagosome membrane>Lipid-anchor. Cytoplasmic vesicle>Autophagosome.
Note: LC3-II binds to the autophagic membranes.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
组织特异性:

Most abundant in heart, brain, liver, skeletal muscle and testis but absent in thymus and peripheral blood leukocytes.

亚基结构:

3 different light chains, LC1, LC2 and LC3, can associate with MAP1A and MAP1B proteins (By similarity). Interacts with TP53INP1 and TP53INP2. Directly interacts with SQSTM1; this interaction leads to MAP1LC3A recruitment to inclusion bodies containing polyubiquitinated protein aggregates and to inclusion body degradation by autophagy. Interacts with ATG13. Interacts with ULK1. Interacts with TBC1D5. Found in a complex with UBQLN1 and UBQLN2. Interacts with UBQLN4 (via STI1 1 and 2 domains). Interacts with UBQLN1 in the presence of UBQLN4. Interacts with TRIM5. Interacts with MEFV. Interacts with RETREG1, RETREG2 and RETREG3. Interacts with PICALM. Interacts with the reticulophagy receptor TEX264.

蛋白家族:

Belongs to the ATG8 family.

功能:

Ubiquitin-like modifier involved in formation of autophagosomal vacuoles (autophagosomes). Plays a role in mitophagy which contributes to regulate mitochondrial quantity and quality by eliminating the mitochondria to a basal level to fulfill cellular energy requirements and preventing excess ROS production. Whereas LC3s are involved in elongation of the phagophore membrane, the GABARAP/GATE-16 subfamily is essential for a later stage in autophagosome maturation. Promotes primary ciliogenesis by removing OFD1 from centriolar satellites via the autophagic pathway. Through its interaction with the reticulophagy receptor TEX264, paticipates in the remodeling of subdomains of the endoplasmic reticulum into autophagosomes upon nutrient stress, which then fuse with lysosomes for endoplasmic reticulum turnover.

翻译修饰:

The precursor molecule is cleaved by ATG4B to form the cytosolic form, LC3-I. This is activated by APG7L/ATG7, transferred to ATG3 and conjugated to phospholipid to form the membrane-bound form, LC3-II.

The Legionella effector RavZ is a deconjugating enzyme that produces an ATG8 product that would be resistant to reconjugation by the host machinery due to the cleavage of the reactive C-terminal glycine.

Phosphorylation at Thr-12 by PKA inhibits conjugation to phosphatidylethanolamine (PE) (By similarity). Interaction with MAPK15 reduces the inhibitory phosphorylation and increases autophagy activity.

细胞定位:

Cytoplasm>Cytoskeleton. Endomembrane system>Lipid-anchor. Cytoplasmic vesicle>Autophagosome membrane>Lipid-anchor. Cytoplasmic vesicle>Autophagosome.
Note: LC3-II binds to the autophagic membranes. Localizes also to discrete punctae along the ciliary axoneme (By similarity).

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
组织特异性:

Most abundant in heart, brain, skeletal muscle and testis. Little expression observed in liver.

亚基结构:

3 different light chains, LC1, LC2 and LC3, can associate with MAP1A and MAP1B proteins (By similarity). Interacts at microtubules with CABP1 (via EF-hands 1 and 2) but not with calmodulin. Interacts with FYCO1 (via C-terminus). Interacts with TP53INP1 and TP53INP2. Interacts with TBC1D25. Directly interacts with SQSTM1; this interaction leads to MAP1LC3B recruitment to inclusion bodies containing polyubiquitinated protein aggregates and to inclusion body degradation by autophagy. Interacts with ATG4B, MAPK15 and BNIP3. Interacts with MAPB1, KEAP1, PCM1, OFD1, CEP131, and TECPR2. Interacts with TBC1D5. Found in a complex with UBQLN1 and UBQLN2. Interacts with UBQLN4 (via STI1 1 and 2 domains). Interacts with UBQLN1 in the presence of UBQLN4. Interacts with ATG13. Interacts with RETREG2, RETREG1 and RETREG3. No interaction, or very weak, with WDFY3. Interacts with PLCL1; the interaction inhibits autophagosome formation (By similarity). Interacts with TRIM16. Interacts with CRY1 and PER2 (By similarity). Interacts with the reticulophagy receptor TEX264.

蛋白家族:

Belongs to the ATG8 family.

研究领域

· Cellular Processes > Cell growth and death > Ferroptosis.   (View pathway)

文献引用

1). Mucin O-glycan-microbiota axis orchestrates gut homeostasis in a diarrheal pig model. Microbiome (PubMed: 36045454) [IF=15.5]

Application: WB    Species: Pig    Sample: colonic tissues

Fig. 6The mechanisms underlying impaired mucin O-glycans in diarrheal piglets. A Immunoblot analysis of the colon of piglets. B The protein levels of NLRP3, ASC, and Caspase 1 in the colon. The concentrations of C IL-1β and D IL-18 in the colonic tissues. E Immunoblot analysis for the colon of piglets. F The protein levels of ATG5, ATG7, LC3A/B, and p62 in the colon. G The mRNA levels of ATG5, ATG7, LC3B, and p62 in the T84 cells after infection with E. coli k88. H The mRNA levels of MUC2, MUC5AC, MUC17, TFF3, RETNLB, FCGBP, and ATF4 in the T84 cells after treatment with 3-MA. Data are presented as mean ± SE. H, healthy controls; D, diarrheal piglets; 3-MA, 3-methyladenine; CON, control group; ETEC, E. coli k88 group; ETEC+3-MA, E. coli k88+3-MA
2). Anthelmintics nitazoxanide protects against experimental hyperlipidemia and hepatic steatosis in hamsters and mice. Acta Pharmaceutica Sinica B (PubMed: 35530137) [IF=14.5]
3). Polysaccharide from Strongylocentrotus nudus eggs regulates intestinal epithelial autophagy through CD36/PI3K-Akt pathway to ameliorate inflammatory bowel disease. International Journal of Biological Macromolecules (PubMed: 37327932) [IF=8.2]
4). A neuroprotective role of Ufmylation through Atg9 in the aging brain of Drosophila. CELLULAR AND MOLECULAR LIFE SCIENCES (PubMed: 37086384) [IF=8.0]
5). d-Borneol enhances cisplatin sensitivity via autophagy dependent EMT signaling and NCOA4-mediated ferritinophagy. PHYTOMEDICINE (PubMed: 36030746) [IF=7.9]
6). Patchouli alcohol protects against chronic unpredictable mild stress-induced depressant-like behavior through inhibiting excessive autophagy via activation of mTOR signaling pathway. BIOMEDICINE & PHARMACOTHERAPY (PubMed: 32244196) [IF=7.5]

Application: WB    Species: rat    Sample: hippocampus

Fig. 6. |The effect of PA on the levels of ex-pression of LC3-II and p62 protein in the hippocampus. (A) The effect of PA on LC3-II and p62 protein levels in hippocampus were in-vestigated by western blot analysis.
7). Inhibition of mTORC1 signaling protects kidney from irradiation-induced toxicity via accelerating recovery of renal stem-like cells. Stem Cell Research & Therapy (PubMed: 30107854) [IF=7.5]

Application: WB    Species: human    Sample: kidney

Fig. 3 | mTORC1 signaling activated by irradiation in kidney tissues.Expression of LC3-I and LC3-II in kidney tissues. Protein lysates prepared from kidney tissues at days 1, 3 and 7 after irradiation. Expression of LC3-I and LC3-II (left panel) detected by western blotting. GAPDH used as a housekeeping control. Nonirradiated kidney tissues (CTL) used as controls. Expression of LC3-I and LC3-II quantitated by ImageJ software and ratio of LC3-II and LC3-I expression presented (right panel). *p < 0.05 vs CTL; ***p < 0.001 vs CTL. CTL nonirradiated renal tissues, GAPDH glyceraldehyde 3-phosphate dehydrogenase, IHC immunohistochemistry, pmTOR pshosphorylated mammalian target of rapamycin, TBI total body irradiation
8). Cladrin alleviates dexamethasone-induced apoptosis of osteoblasts and promotes bone formation through autophagy induction via AMPK/mTOR signaling. Free Radical Biology and Medicine (PubMed: 35998794) [IF=7.4]
9). YAP1 alleviates sepsis-induced acute lung injury via inhibiting ferritinophagy-mediated ferroptosis. Frontiers in Immunology (PubMed: 35979359) [IF=7.3]

Application: WB    Species: Mice    Sample: lung tissues

Figure 7 YAP1 deficiency aggravated CLP-induced ferroptosis and ferritinophagy in lung tissues. (A) Representative images shown by TEM. The red arrow indicates representative mitochondria in WT and YAP1f/f mice lungs treated by CLP or not (n = 3 mice/group). (B) Fluorescence analysis showed representative images of ferritin colocalization (green) with tissue expression of LC3 (red) and quantification fluorescence intensity of LC3. Nuclei were counterstained by DAPI (blue). Light pink fluorescence represented the colocalization of ferritin and LC3. (C–F) Immunoblotting detection of the expression contents of GPX4 (D), SLC7A11 (E), and SFXN1 (F) in mice with indicated treatment. (G–J) Immunoblotting determination of the expression levels of LC3II/LC3I (H), NCOA4 (I), and FTH1 (J) in mice with equivalent treatment. Data are expressed as mean ± SD. n = 6; *p < 0.05, **p < 0.01, ***p < 0.001.

Application: IHC    Species: Mice    Sample: lung tissues

Figure 7 YAP1 deficiency aggravated CLP-induced ferroptosis and ferritinophagy in lung tissues. (A) Representative images shown by TEM. The red arrow indicates representative mitochondria in WT and YAP1f/f mice lungs treated by CLP or not (n = 3 mice/group). (B) Fluorescence analysis showed representative images of ferritin colocalization (green) with tissue expression of LC3 (red) and quantification fluorescence intensity of LC3. Nuclei were counterstained by DAPI (blue). Light pink fluorescence represented the colocalization of ferritin and LC3. (C–F) Immunoblotting detection of the expression contents of GPX4 (D), SLC7A11 (E), and SFXN1 (F) in mice with indicated treatment. (G–J) Immunoblotting determination of the expression levels of LC3II/LC3I (H), NCOA4 (I), and FTH1 (J) in mice with equivalent treatment. Data are expressed as mean ± SD. n = 6; *p < 0.05, **p < 0.01, ***p < 0.001.
10). Repurposing nitazoxanide as a novel anti‐atherosclerotic drug based on mitochondrial uncoupling mechanisms. British Journal of Pharmacology (PubMed: 36082580) [IF=7.3]
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