Key Product Details

Validated by

Tagged Protein Expression

Species Reactivity

Validated:

Human, Mouse, Rat, Porcine, Bacteria, Bovine, Canine, Primate, Yeast, Zebrafish

Cited:

Human, Mouse, Rat, Porcine, Bacteria, Bovine, Canine, Fish - Danio rerio (Zebrafish), Primate

Applications

Validated:

Electron Microscopy, Flow Cytometry, Immunoblotting, Immunocytochemistry/Immunofluorescence, Immunohistochemistry, Immunohistochemistry Free-Floating, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Immunoprecipitation, Knockdown Validated, Knockout Validated, Simple Western, Western Blot

Cited:

Block/Neutralize, Electron Microscopy, Flow Cytometry, IF/IHC, Immunoblotting, Immunocytochemistry, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry Free-Floating, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Immunoprecipitation, In vivo assay, Knockdown Validated, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free

Concentration

1.0 mg/ml

View all LC3B Antibodies »

Product Summary for LC3B Antibody - BSA Free

Immunogen

Polyclonal LC3B Antibody was made to a synthetic peptide made to the N-terminal region of the human LC3B protein.

Reactivity Notes

Use in Yeast reported in scientific literature (PMID:35247568).Use in Rat reported in scientific literature (PMID:34499623). Mouse reactivity reported in scientific literature (PMID:32802192). Zebrafish reactivity reported in scientific literature (PMID: 23724125). . Canine and primate reactivity reported in scientific literature (PMID: 24027311). Porcine reactivity reported in scientific literature (PMID: 25378587). . Rat reactivity reported in scientific literature (30067379). . Bacteria reactivity reported in scientific literature (31110360). . Bovine reactivity reported in scientific literature (21868124). . Other species have not been tested.

Localization

LC3-I is cytoplasmic. LC3-II binds to the autophagic membranes.

Marker

Autophagosome Marker

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

14.688 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for LC3B Antibody - BSA Free

Western Blot Detection of LC3B in Transfected FTC-133 and TPC-1 Cells with Various Treatments

Western Blot Detection of LC3B in Transfected FTC-133 and TPC-1 Cells with Various Treatments

FTC-133/TPC-1 cells were transfected with HMGB1 shRNA and control shRNA and then pre-treated for 1 h with pepstatin A (PA, 10 uM) and E64D (10 uM) as indicated. Cells were subsequently treated for 3 h with HBSS in continuous presence or absence PA/E64D inhibitors. LC3-I/II levels were assayed by Western blot. Image collected and cropped by CiteAb from the following publication (https://jeccr.biomedcentral.com/articles/10.1186/s13046-019-1328-3) licensed under a CC-BY licence.
Immunocytochemistry/Immunofluorescence Staining of LC3B in HeLa Cells

Immunocytochemistry/Immunofluorescence Staining of LC3B in HeLa Cells

Analysis of LC3B in HeLa cells using anti-LC3B antibody (red) [Catalog # NB600-1384]. Nuclei were counterstained with DAPI (blue).
Immunohistochemical Staining of LC3B in CC Tissue with Low and High LC3B Dot Like Staining

Immunohistochemical Staining of LC3B in CC Tissue with Low and High LC3B Dot Like Staining

LC3B immunohistochemical staining in CC tissue(A) CC with low LC3B dot like staining. (B) CC with high LC3B dot like staining. (magnification 20X). Image collected and cropped by CiteAb from the following publication (https://www.oncotarget.com/lookup/doi/10.18632/oncotarget.17554) licensed under a CC-BY licence.
View 13 more images

Applications for LC3B Antibody - BSA Free

Application
Recommended Usage

Flow Cytometry

1:200

Immunohistochemistry-Frozen

1:400

Immunohistochemistry-Paraffin

1:200-1:400

Simple Western

1:100

Western Blot

1:1000
Application Notes
In Western Blot, bands are seen at ~17 and 19 kDa corresponding to LC3-II and LC3-I. The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors. Electron Microscopy was reported in scientific literature (PMID: 21885071). Use in Immunohistochemistry-free floating reported in scientific literature (PMID: 24928515). Use in Immunohistochemistry on both paraffin-embedded and frozen sections reported in scientific literature (PMID: 18259115). Use in immunoprecipitation reported in scientific literature (PMID: 26098573). Use in immunoblotting reported in scientific literature (PMID: 25383539). Use in flow reported in scientific literature (PMID: 27622036). In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue.
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 18 reviews rated 4.2 using NB600-1384 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.

Background: LC3B

Autophagy (macroautophagy) is a catabolic process which targets intracellular components such as proteins and organelles for degradation. Originally described as a bulk degradation process, current research supports its selective nature (1). Selective autophagy targets specific cellular components for degradation including the endoplasmic reticulum (2) (ER-phagy), mitochondria (3) (mitophagy), peroxisomes (3) (pexophagy), ribosomes (4) (ribophagy) and bacteria (5) (xenophagy). Autophagy relies on a newly formed phagophore, a membrane structure which elongates, sequesters cellular content, and fuses to form a double membrane vesicle known as the autophagosome. Fusion of autophagosomes with lysosomes gives rise to the autophagolysosome, where cellular components are degraded by lysosome hydrolases (1).

Autophagic flux is supported by autophagy-related proteins (Atgs) initially identified in yeast (6,7). The core autophagy machinery is comprised of 17 Atg proteins that play specific roles in autophagosome formation. Among these Atg proteins, Atg8 is not only involved in autophagosome formation but also functions in cargo selection. In mammals, several Atg8 homologues have been identified including microtubule-associated protein 1 light chain 3 alpha, beta and gamma - LC3A, LC3B, and LC3C (8) respectively, as well as GABA type A receptor-associated protein (GABARAP), GABARAP-Like1, and GABARAP-Like2 (9). LC3 (predicted molecular weight 14kD) is ubiquitously expressed and undergoes posttranslational processing after synthesis. First, the cysteine protease Atg4 cleaves a carboxy terminal sequence to generate the cytosolic form LC3-I. Next, E1-like (Atg7) and E2-like (Atg3) enzymes conjugate phosphatidylethanolamine to the newly exposed carboxyterminal glycine, generating LC3-II. Finally, the Atg12-Atg5-Atg16L1 complex participates in LC3 lipidation and autophagosome formation (10). LC3B-I to LC3B-II conversion correlates with autophagosome number and is considered the best marker to monitor autophagy.

References

1. Yu, L., Chen, Y., & Tooze, S. A. (2018). Autophagy pathway: Cellular and molecular mechanisms. Autophagy. https://doi.org/10.1080/15548627.2017.1378838

2. Forrester, A., De Leonibus, C., Grumati, P., Fasana, E., Piemontese, M., Staiano, L., ... Settembre, C. (2019). A selective ER -phagy exerts procollagen quality control via a Calnexin- FAM 134B complex. The EMBO Journal. https://doi.org/10.15252/embj.201899847

3. He, X., Zhu, Y., Zhang, Y., Geng, Y., Gong, J., Geng, J., ... Zhong, H. (2019). RNF34 functions in immunity and selective mitophagy by targeting MAVS for autophagic degradation. The EMBO Journal. https://doi.org/10.15252/embj.2018100978

4. Mathai, B., Meijer, A., & Simonsen, A. (2017). Studying Autophagy in Zebrafish. Cells. https://doi.org/10.3390/cells6030021

5. Losier, T. T., Akuma, M., McKee-Muir, O. C., LeBlond, N. D., Suk, Y., Alsaadi, R. M., ... Russell, R. C. (2019). AMPK Promotes Xenophagy through Priming of Autophagic Kinases upon Detection of Bacterial Outer Membrane Vesicles. Cell Reports. https://doi.org/10.1016/j.celrep.2019.01.062

6. Nakatogawa, H., Suzuki, K., Kamada, Y., & Ohsumi, Y. (2009). Dynamics and diversity in autophagy mechanisms: Lessons from yeast. Nature Reviews Molecular Cell Biology. https://doi.org/10.1038/nrm2708

7. Tsukada, M., & Ohsumi, Y. (1993). Isolation and characterization of autophagy-defective mutants of Saccharomyces cerevisiae. FEBS Letters. https://doi.org/10.1016/0014-5793(93)80398-E

8. Wild, P., McEwan, D. G., & Dikic, I. (2014). The LC3 interactome at a glance. Journal of Cell Science. https://doi.org/10.1242/jcs.140426

9. Igloi, G. L. (2001). Cloning, expression patterns, and chromosome localization of three human and two mouse homologues of GABAA receptor-associated protein. Genomics. https://doi.org/10.1006/geno.2001.6555

10. Glick, D., Barth, S., & Macleod, K. F. (2010). Autophagy: Cellular and molecular mechanisms. Journal of Pathology. https://doi.org/10.1002/path.2697

Long Name

Microtubule-associated Protein 1 Light Chain 3 beta

Alternate Names

Apg8b, ATG8F, LC3II, MAP1LC3B

Gene Symbol

MAP1LC3B

Additional LC3B Products

Product Documents for LC3B Antibody - BSA Free

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Product Specific Notices for LC3B Antibody - BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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