LC3A/B Antibody - #AF5402

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製品: LC3A/B Antibody
カタログ: AF5402
タンパク質の説明: Rabbit polyclonal antibody to LC3A/B
アプリケーション: WB IHC IF/ICC
Cited expt.: WB, IHC, IF/ICC
反応性: Human, Mouse, Rat
予測: Pig, Zebrafish, Bovine, Sheep, Dog, Xenopus
分子量: 14kDa,16 kDa; 14kD,15kD(Calculated).
ユニプロット: Q9H492 | Q9GZQ8
RRID: AB_2837886

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WB Handbook
IHC Protocol
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製品説明

ソース:
Rabbit
アプリケーション:
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: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

反応性:
Human,Mouse,Rat
予測:
Pig(100%), Zebrafish(93%), Bovine(100%), Sheep(100%), Dog(100%), Xenopus(100%)
クローナリティ:
Polyclonal
特異性:
LC3A/B Antibody detects endogenous levels of total LC3A/B.
RRID:
AB_2837886
引用形式: Affinity Biosciences Cat# AF5402, RRID:AB_2837886.
コンジュゲート:
Unconjugated.
精製:
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.
別名:

折りたたみ／展開

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;

免疫原

免疫原:

A synthesized peptide derived from human LC3B, corresponding to a region within N-terminal amino acids.

Uniprot:

Q9H492(MLP3A_HUMAN) >>Visit HPA database.

Q9GZQ8(MLP3B_HUMAN) >>Visit HPA database.

遺伝子(ID):
MAP1LC3A(84557) | MAP1LC3B(81631)
発現特異性:
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.

タンパク質の説明:
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.
タンパク質配列:
MPSDRPFKQRRSFADRCKEVQQIRDQHPSKIPVIIERYKGEKQLPVLDKTKFLVPDHVNMSELVKIIRRRLQLNPTQAFFLLVNQHSMVSVSTPIADIYEQEKDEDGFLYMVYASQETFGF

MPSEKTFKQRRTFEQRVEDVRLIREQHPTKIPVIIERYKGEKQLPVLDKTKFLVPDHVNMSELIKIIRRRLQLNANQAFFLLVNGHSMVSVSTPISEVYESEKDEDGFLYMVYASQETFGMKLSV

種類予測

種類予測:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

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

研究背景

機能:

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.

PTMs:

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.

細胞の位置付け:

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
組織特異性:

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

タンパク質ファミリー:

Belongs to the ATG8 family.

機能:

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.

PTMs:

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.

細胞の位置付け:

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
組織特異性:

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

タンパク質ファミリー:

Belongs to the ATG8 family.

研究領域

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

参考文献

1). Anthelmintics nitazoxanide protects against experimental hyperlipidemia and hepatic steatosis in hamsters and mice. Acta Pharmaceutica Sinica B, 2022 (PubMed: 35530137) [IF=14.7]
2). CENPN suppresses autophagy and increases paclitaxel resistance in nasopharyngeal carcinoma cells by inhibiting the CREB-VAMP8 signaling axis. Autophagy, 2024 (PubMed: 37776538) [IF=14.6]

Application: WB    Species: Human    Sample: NPC cells

Figure 2. CENPN expression affects autophagy and PTX resistance in NPC cells. (A) The cytotoxicity assay showed that knockdown of CENPN reduced PTX resistance in NPC cells. (B) The cytotoxicity assay showed that overexpression of CENPN enhanced PTX resistance in NPC cells. (C) The CCK8 assay showed the effect of PTX (10 nM) on the viability of NPC cells after knockdown of CENPN. (D) The CCK8 assay showed the effect of PTX (5 nM) on the viability of NPC cells after overexpression of CENPN. (E, F) WB analysis showed the effect of PTX (10 nM) on the levels of autophagy in NPC cells after knockdown of CENPN. (G, H) WB analysis showed the effect of PTX (5 nM) on the levels of autophagy in NPC cells after overexpression of CENPN. Data are presented as mean ± SD.
3). Mucin O-glycan-microbiota axis orchestrates gut homeostasis in a diarrheal pig model. Microbiome, 2022 (PubMed: 36045454) [IF=13.8]

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
4). Rationally designed catalytic nanoplatform for enhanced chemoimmunotherapy via deploying endogenous plus exogenous copper and remodeling tumor microenvironment. Journal of nanobiotechnology, 2024 (PubMed: 39252079) [IF=10.2]

Application: WB    Species: Mouse    Sample:

Figure 3. Synergistic tumor inhibition of CP and CQ dependent on ROS generation and irrespective of autophagosome accumulation. (A) Tumor volume curves were monitored during different treatments by intratumoral injection. CP, 5 mg/kg; CQ, 8.56 mg/kg; wortmannin (Wort), 0.35 mg/kg. n=5. (B and C) Photograph images and average tumor weights of the dissected tumors after the indicated treatments. n=5. (Dand E) Western blot analysis (D) of CT-26 cells exposed to the indicated treatments in the presence or absence of ATG5 expression attenuation. Image J software was used (E). CP, 15 μg/mL; CQ, 50 μM. Triple individual experiments were performed. (F) Cell viability analysis of CT-26 cells after the indicated treatments for 24 h in the presence or absence of ATG5 expression attenuation. CP, 15 μg/mL; CQ, 50 μM. Triple individual experiments were performed. n=3. (G and H) ROS analysis by flow cytometry after CT-26 after exposed to different treatments for 24 h and then stained with DCFH-DA (10 μM; 20 min). Triple individual experiments were performed. (I and J) Cell viability analysis (24 h treatment) and ROS analysis (6 h treatment) of CT-26 cells after the indicated treatments. CP, 15 μg/mL; CQ, 50 μM; NAC, 5 mM. (K) Total copper contents in CT-26 cells after the indicated treatments for 12 h was quantified by ICP-OES. CQ, 50 μM. (L) Fluorescence images stained by R6G (10 μΜ; 20 min) for CT-26 after the indicated treatments for 6 h. CP, 15 μg/mL; CQ, 50 μΜ. λex= 495 nm. (M) Western blot analysisof CT-26 cells after the indicated treatments for 24 h. CP, 15 μg/mL; CQ, 50 μM.
5). Autophagy-mediated activation of the AIM2 inflammasome enhances M1 polarization of microglia and exacerbates retinal neovascularization. MedComm, 2024 (PubMed: 39081514) [IF=9.9]

Application: WB    Species: Mouse    Sample: normoxic and hypoxic BV2 cells

FIGURE 3. Autophagy regulates the protein level of absent in melanoma 2 (AIM2). (A and B) Reactive oxygen species (ROS) levels of BV2 cells under normoxia or hypoxia (mean ± standard deviation [SD]; n = 5/group; ** p < 0.01, unpaired Student's t‐test; scale bar: 200 µm). (C and D) Retinal neovascularization (RNV) area in oxygen‐induced retinopathy (OIR) with DMSO or GKT831 (mean ± SD; n = 4/group; ** p < 0.01, unpaired Student's t‐test; scale bar: 1 mm). (E) Images of transmission electron microscopy (TEM) between normoxic and hypoxic BV2 cells. Scale bar: 1 µm, 200 nm. (F) Protein expression and quantification of autophagy marker LC3B in normoxic and hypoxic BV2 cells (mean ± SD; n = 3/group; * p < 0.05, ** p < 0.01, unpaired Student's t‐test). (G) Expression and quantification of AIM2 after disposing chloroquine at concentrations of 5, 10, 20, 40, and 80 µm (mean ± SD; n = 3/group; ** p < 0.001, one‐way analysis of variance [ANOVA]). (H) Protein expression and quantification of AIM2 in hypoxic BV2 cells treated with rapamycin (mean ± SD; n = 3/group; * p < 0.05, one‐way ANOVA). (I) Co‐immunoprecipitation (Co‐IP) of AIM2 with LC3B or P62. DMSO, Dimethyl Sulfoxide.
6). Polysaccharide from Strongylocentrotus nudus eggs regulates intestinal epithelial autophagy through CD36/PI3K-Akt pathway to ameliorate inflammatory bowel disease. International Journal of Biological Macromolecules, 2023 (PubMed: 37327932) [IF=7.7]
7). Inhibition of mTORC1 signaling protects kidney from irradiation-induced toxicity via accelerating recovery of renal stem-like cells. Stem Cell Research & Therapy, 2018 (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, 2022 (PubMed: 35998794) [IF=7.1]
9). Patchouli alcohol protects against chronic unpredictable mild stress-induced depressant-like behavior through inhibiting excessive autophagy via activation of mTOR signaling pathway. BIOMEDICINE & PHARMACOTHERAPY, 2020 (PubMed: 32244196) [IF=6.9]

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.
10). Repurposing nitazoxanide as a novel anti‐atherosclerotic drug based on mitochondrial uncoupling mechanisms. British Journal of Pharmacology, 2023 (PubMed: 36082580) [IF=6.8]
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