LC3B Antibody - #AF4650

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Product: LC3B Antibody
Catalog: AF4650
Description: Rabbit polyclonal antibody to LC3B
Application: WB IHC IF/ICC
Cited expt.: WB, IHC, IF/ICC
Reactivity: Human, Mouse, Rat
Prediction: Pig, Zebrafish, Bovine, Sheep, Dog, Xenopus
Mol.Wt.: 14kDa,16kDa; 15kD(Calculated).
Uniprot: Q9GZQ8
RRID: AB_2844592

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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, IF/ICC 1:100-1:500, IHC 1:50-1:200
*The optimal dilutions should be determined by the end user. For optimal experimental results, antibody reuse is not recommended.
*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(92%), Bovine(100%), Sheep(100%), Dog(100%), Xenopus(100%)
Clonality:
Polyclonal
Specificity:
LC3B Antibody detects endogenous levels of total LC3B.
RRID:
AB_2844592
Cite Format: Affinity Biosciences Cat# AF4650, RRID:AB_2844592.
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

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:

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

Uniprot:

Q9GZQ8(MLP3B_HUMAN) >>Visit HPA database.

Gene(ID):
MAP1LC3B(81631)
Expression:
Q9GZQ8 MLP3B_HUMAN:

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

Sequence:
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
92
Horse
0
Chicken
0
Rabbit
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

Research Backgrounds

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:

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.

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. 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
Tissue Specificity:

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

Family&Domains:

Belongs to the ATG8 family.

Research Fields

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

References

1). A Sirtuin-1-Targeted Gene-Activating Tetrahedral DNA Attenuates Bladder Fibrosis by Restoring Mitophagy in Fibroblasts via the SIRT1-FOXO3-BNIP3 Axis. Advanced science (Weinheim, Baden-Wurttemberg, Germany), 2025 (PubMed: 41276935) [IF=15.1]
2). Endothelial F3-mediated autolysosome and lipid metabolism promote resistance to anti-VEGFA therapy in metastatic colorectal cancer. Autophagy, 2025 (PubMed: 40922383) [IF=14.6]
3). The NLRX1-SLC39A7 complex orchestrates mitochondrial dynamics and mitophagy to rejuvenate intervertebral disc by modulating mitochondrial Zn2+ trafficking. Autophagy, 2024 (PubMed: 37876250) [IF=14.6]

Application: WB    Species: human    Sample: NP cells

Figure 3. NLRX1 is essential for the beneficial action of mitophagy in alleviating mitochondrial dysfunction and NP cell senescence. Primary human NP cells isolated from NP tissues with different Pfirrmann grades were prepared. (A) confocal analysis of NLRX1 and MAP1LC3B with if staining in NP cells isolated from health NP tissues treated by PBS or TBHP, scale bar: 10 μm. (B) proteins immunoprecipitated (IP) from NP cells isolated from health NP tissues treated by PBS or TBHP followed by western blotting, left panel: IP with anti-MAP1LC3B antibody, right panel: IP with anti-NLRX1 antibody. (C) confocal analysis of TOMM20 and MAP1LC3B with if staining in NP cells isolated from degenerated NP tissues treated by PBS or TBHP with NLRX1 overexpression or not, scale bar: 10 μm. (D and E) protein expressions of mitophagy indicators (MAP1LC3B-II, TOMM20, TIMM23) in primary human NP cells isolated from degenerated NP tissues treated by PBS or TBHP with NLRX1 overexpression or not, as determined by western blotting. (F-I) JC-1 incubation for detecting mitochondrial membrane potential (MMP; F, H) and DCFH incubation for detecting reactive oxidative species (ROS; G, I) in NP cells isolated from degenerated NP tissues treated by PBS, TBHP or 3-MA with NLRX1 overexpression or not, as determined by flow cytometry. (J and K) protein expressions of senescence indicators (TP53, CDKN2A) and SASP factors (IL1B, IL6) in NP cells isolated from degenerated NP tissues treated by PBS, TBHP or 3-MA with NLRX1 overexpression or not, as determined by western blotting. (L and M) cell proliferation (MKI67 immunofluorescent staining, EdU incorporation) and cell senescence (SA-GLB1/β-gal staining) in primary human NP cells isolated from degenerated NP tissues treated by PBS, TBHP or 3-MA with NLRX1 overexpression or not, scale bar: 50 μm (IF images), 100 μm (white light images). Data are represented as mean ± SD.

Application: IF/ICC    Species: human    Sample: NP cells

Figure 3. NLRX1 is essential for the beneficial action of mitophagy in alleviating mitochondrial dysfunction and NP cell senescence. Primary human NP cells isolated from NP tissues with different Pfirrmann grades were prepared. (A) confocal analysis of NLRX1 and MAP1LC3B with if staining in NP cells isolated from health NP tissues treated by PBS or TBHP, scale bar: 10 μm. (B) proteins immunoprecipitated (IP) from NP cells isolated from health NP tissues treated by PBS or TBHP followed by western blotting, left panel: IP with anti-MAP1LC3B antibody, right panel: IP with anti-NLRX1 antibody. (C) confocal analysis of TOMM20 and MAP1LC3B with if staining in NP cells isolated from degenerated NP tissues treated by PBS or TBHP with NLRX1 overexpression or not, scale bar: 10 μm. (D and E) protein expressions of mitophagy indicators (MAP1LC3B-II, TOMM20, TIMM23) in primary human NP cells isolated from degenerated NP tissues treated by PBS or TBHP with NLRX1 overexpression or not, as determined by western blotting. (F-I) JC-1 incubation for detecting mitochondrial membrane potential (MMP; F, H) and DCFH incubation for detecting reactive oxidative species (ROS; G, I) in NP cells isolated from degenerated NP tissues treated by PBS, TBHP or 3-MA with NLRX1 overexpression or not, as determined by flow cytometry. (J and K) protein expressions of senescence indicators (TP53, CDKN2A) and SASP factors (IL1B, IL6) in NP cells isolated from degenerated NP tissues treated by PBS, TBHP or 3-MA with NLRX1 overexpression or not, as determined by western blotting. (L and M) cell proliferation (MKI67 immunofluorescent staining, EdU incorporation) and cell senescence (SA-GLB1/β-gal staining) in primary human NP cells isolated from degenerated NP tissues treated by PBS, TBHP or 3-MA with NLRX1 overexpression or not, scale bar: 50 μm (IF images), 100 μm (white light images). Data are represented as mean ± SD.
4). Sirt3-mediated mitophagy regulates AGEs-induced BMSCs senescence and senile osteoporosis. Redox Biology, 2021 (PubMed: 33662874) [IF=10.7]

Application: WB    Species: mice    Sample: bone marrow mesenchymal stem (BMSCs)

Fig. 2. Effects of different concentrations of AGEs on mitochondrial function and mitophagy of BMSCs. The BMSCs were treated with AGEs (50–200 μg/mL) or BSA for 24–72 h. (A) Representative fluorescence images with DCF (green) staining in BMSCs stimulated with AGEs. Scale bar: 50 μm. (B) Representative fluorescence images with Mito-SOX (red) and Mito-Tracker (green) double-staining in BMSCs stimulated with AGEs. Scale bar: 50 μm. (C) The MMP was detected through JC-1 staining in BMSCs stimulated with AGEs. Scale bar: 50 μm. (D) Representative fluorescence images with Mtphagy Dye (red) and Mito-Tracker (green) double-staining in BMSCs stimulated with AGEs. Scale bar: 50 μm. (E) Representative fluorescence images with LC3B (red) and Mito-Tracker (green) double-staining in BMSCs stimulated with AGEs. Scale bar: 50 μm. (F) Representative Western blotting assay and quantitation of the level of LC3B, P62, Parkin, Sirt3. **p < 0.01 versus BSA. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)
5). METTL3-mediated m6A modification of SIRT1 mRNA modulates podocyte autophagy in diabetes nephropathy with the treatment effect of Astragalus polysaccharide. International journal of biological macromolecules, 2025 (PubMed: 41349753) [IF=7.7]
6). Pentapeptide PYRAE triggers ER stress-mediated apoptosis of breast cancer cells in mice by targeting RHBDF1-BiP interaction. Acta pharmacologica Sinica, 2024 (PubMed: 37798352) [IF=6.9]
7). Tert-butylhydroquinone promotes skin flap survival by inhibiting oxidative stress mediated by the Nrf2/HO-1 signalling pathway.. British journal of pharmacology, 2024 (PubMed: 39233316) [IF=6.8]
8). Inhibition of GSK3β Reduces Ectopic Lipid Accumulation and Induces Autophagy by the AMPK Pathway in Goat Muscle Satellite Cells. Cells, 2019 (PubMed: 31683987) [IF=6.0]
9). Effect of mitophagy in the formation of osteomorphs derived from osteoclasts. iScience, 2023 (PubMed: 37250312) [IF=5.8]
10). TAAR1 deficiency impairs mitochondrial dynamics and synaptic integrity in the medial prefrontal cortex and associated cognition in mice. Translational psychiatry, 2025 (PubMed: 41271642) [IF=5.8]
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