LC3A/B Antibody | Affinity Biosciences

Product:	LC3A/B Antibody
Catalog:	AF5402
Description:	Rabbit polyclonal antibody to LC3A/B
Application:	WB IHC IF/ICC
Reactivity:	Human, Mouse, Rat
Prediction:	Pig, Zebrafish, Bovine, Sheep, Dog, Xenopus
Mol.Wt.:	14kDa,16 kDa; 14kD,15kD(Calculated).
Uniprot:	Q9H492 \| Q9GZQ8
RRID:	AB_2837886

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Blocking peptides are peptides that bind specifically to the target antibody and block antibody binding. These peptide usually contains the epitope recognized by the antibody. Antibodies bound to the blocking peptide no longer bind to the epitope on the target protein. By comparing the staining from the blocked antibody versus the antibody alone, one can see which staining is specific.

Size	Price	Inventory
100ul	$280	In stock
200ul	$350	In stock

Lead Time: Same day delivery

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Related Downloads

MSDS

Data Sheet

COA

Protocols

WB Handbook

IHC Protocol

IF/ICC Protocol

Product Info

Source:

Rabbit

Application:

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.

Reactivity:

Human,Mouse,Rat

Prediction:

Pig(100%), Zebrafish(93%), Bovine(100%), Sheep(100%), Dog(100%), Xenopus(100%)

Clonality:

Polyclonal

Specificity:

LC3A/B Antibody detects endogenous levels of total LC3A/B.

RRID:

AB_2837886
Cite Format: Affinity Biosciences Cat# AF5402, RRID:AB_2837886.

Conjugate:

Unconjugated.

Purification:

The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).

Storage:

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.

Alias:

Fold/Unfold

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;

Immunogens

Immunogen:

Uniprot:

Q9H492(MLP3A_HUMAN) >>Visit HPA database.

Q9GZQ8(MLP3B_HUMAN) >>Visit HPA database.

Gene(ID):

MAP1LC3A(84557) | MAP1LC3B(81631)

Expression:

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.

Description:

Sequence:

Q9H492 MLP3A_HUMAN:
Protein BLAST With
NCBI/
ExPASy/
Uniprot

MPSDRPFKQRRSFADRCKEVQQIRDQHPSKIPVIIERYKGEKQLPVLDKTKFLVPDHVNMSELVKIIRRRLQLNPTQAFFLLVNQHSMVSVSTPIADIYEQEKDEDGFLYMVYASQETFGF

Q9GZQ8 MLP3B_HUMAN:
Protein BLAST With
NCBI/
ExPASy/
Uniprot

MPSEKTFKQRRTFEQRVEDVRLIREQHPTKIPVIIERYKGEKQLPVLDKTKFLVPDHVNMSELIKIIRRRLQLNANQAFFLLVNGHSMVSVSTPISEVYESEKDEDGFLYMVYASQETFGMKLSV

Predictions

Predictions:

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

Horse

Chicken

Rabbit

Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - Q9H492/Q9GZQ8 As Substrate

Q9H492
Q9GZQ8

Site	PTM Type	Enzyme	Source
S12	Phosphorylation	P17612 (PRKACA)	Uniprot
K42	Ubiquitination		Uniprot
K49	Acetylation		Uniprot
K49	Ubiquitination		Uniprot
T50	Phosphorylation	Q13188 (STK3) , Q13043 (STK4)	Uniprot
K51	Acetylation		Uniprot
S92	Phosphorylation		Uniprot
T93	Phosphorylation		Uniprot

Site	PTM Type	Source
K5	Ubiquitination	Uniprot
T6	Phosphorylation	Uniprot
R21	Methylation	Uniprot
T29	Phosphorylation	Uniprot
K30	Ubiquitination	Uniprot
K42	Ubiquitination	Uniprot
K49	Ubiquitination	Uniprot
K51	Ubiquitination	Uniprot
K65	Ubiquitination	Uniprot

Research Backgrounds

Q9H492_MLP3A_HUMAN
Q9GZQ8_MLP3B_HUMAN

Function:

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.

Subcellular Location:

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

Tissue Specificity:

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

Subunit Structure:

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.

Family&Domains:

Belongs to the ATG8 family.

Function:

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:

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

Subcellular Location:

Tissue Specificity:

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

Subunit Structure:

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.

Family&Domains:

Belongs to the ATG8 family.

Research Fields

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

References

1). Mucin O-glycan-microbiota axis orchestrates gut homeostasis in a diarrheal pig model. Microbiome, 2022 (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, 2022 (PubMed: 35530137) [IF=14.5]

3). CENPN suppresses autophagy and increases paclitaxel resistance in nasopharyngeal carcinoma cells by inhibiting the CREB-VAMP8 signaling axis. Autophagy, 2024 (PubMed: 37776538) [IF=13.3]

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.

4). Autophagy-mediated activation of the AIM2 inflammasome enhances M1 polarization of microglia and exacerbates retinal neovascularization. MedComm, 2024 (PubMed: 39081514) [IF=10.7]

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.

5). 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]

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=8.2]

7). A neuroprotective role of Ufmylation through Atg9 in the aging brain of Drosophila. CELLULAR AND MOLECULAR LIFE SCIENCES, 2023 (PubMed: 37086384) [IF=8.0]

8). d-Borneol enhances cisplatin sensitivity via autophagy dependent EMT signaling and NCOA4-mediated ferritinophagy. PHYTOMEDICINE, 2022 (PubMed: 36030746) [IF=7.9]

9). 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

10). 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=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.

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LC3A/B Antibody - #AF5402