- Datasheet
- Specific References (140)
- Protocols
Overview
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Product nameAnti-LC3B antibody
See all LC3B primary antibodies -
DescriptionRabbit polyclonal to LC3B
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Host speciesRabbit
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Tested applicationsSuitable for: ICC/IF, IP, IHC-P, WB, IHC-Frmore details
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Species reactivityReacts with: Mouse, Rat, Dog, Human, Zebrafish, Syrian hamster, Bacteria
Predicted to work with: Cow
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Immunogen
Synthetic peptide corresponding to Human LC3B (N terminal). A synthetic peptide made to an N-terminal portion of the human LC3 protein sequence (between residues 1-100).
Database link: Q9GZQ8 -
Positive control
- Human brain lysate, mouse brain lysate, ES, HeLa, HEK293 and MCF7 cells, human lymphocytes and embryoid bodi.
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
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Storage bufferPreservative: 0.02% Sodium azide
Constituent: PBS -
Concentration information loading... -
PurityImmunogen affinity purified
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ClonalityPolyclonal
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IsotypeIgG
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Research areas
Associated products
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Compatible Secondaries
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Immunohistochemistry kits
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Related Products
Applications
Our Abpromise guarantee covers the use of ab48394 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| ICC/IF | Use a concentration of 1 µg/ml. | |
| IP | Use at an assay dependent concentration. 20 ug per 500 ug of protein. | |
| IHC-P | 1/200 - 1/400. | |
| WB | Use a concentration of 0.5 - 2 µg/ml. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa). Detects bands of approximately 17 kDa (LC3-II) and 19 kDa (LC3-I) (predicted molecular weight: 15 kDa). |
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| IHC-Fr | Use at an assay dependent concentration. |
Target
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FunctionProbably involved in formation of autophagosomal vacuoles (autophagosomes).
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Tissue specificityMost abundant in heart, brain, skeletal muscle and testis. Little expression observed in liver.
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Sequence similaritiesBelongs to the MAP1 LC3 family.
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Post-translational
modificationsThe precursor molecule is cleaved by APG4B/ATG4B to form LC3-I. This is activated by APG7L/ATG7, transferred to ATG3 and conjugated to phospholipid to form LC3-II. -
Cellular localizationCytoplasm > cytoskeleton. Endomembrane system. Cytoplasmic vesicle > autophagosome membrane. LC3-II binds to the autophagic membranes.
- Information by UniProt
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Database links
- Entrez Gene: 408001 Cow
- Entrez Gene: 81631 Human
- Entrez Gene: 67443 Mouse
- Entrez Gene: 64862 Rat
- Entrez Gene: 322425 Zebrafish
- Omim: 609604 Human
- SwissProt: O41515 Cow
- SwissProt: Q9GZQ8 Human
see all -
Alternative names
- ATG8F antibody
- Autophagy-related protein LC3 B antibody
- Autophagy-related ubiquitin-like modifier LC3 B antibody
see all
Images
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Western blot - Anti-LC3B antibody (ab48394)All lanes : Anti-LC3B antibody (ab48394) at 2 µg/ml
Lane 1 : ATG5+ control lysate (see reference)
Lane 2 : ATG5- control lysate (see reference)
Lysates/proteins at 40 µg per lane.
Secondary
All lanes : Goat anti-rabbit HRP conjugated
Developed using the ECL technique.
Predicted band size: 15 kDa
Observed band size: 17,19 kDa (why is the actual band size different from the predicted?)
Additional bands at: 38 kDa, 40 kDa, 80 kDa. We are unsure as to the identity of these extra bands.
Observed band at 18 kDa: LC3-I.
Observed band at 17 kDa: LC3-II. -
Immunocytochemistry/ Immunofluorescence - Anti-LC3B antibody (ab48394)This image is courtesy of an Abreview by Armen Petrosyan.ab48394 staining LC3B in a Rat hepatocyte by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde, permeabilized with 0.2% Triton X-100 in PBS and blocked with 1% Donkey serum in 0.1% PBST for 60 minutes at 21°C. Samples were incubated with primary antibody (1/50 in PBS + 1% BSA) for 3 hours at 22°C. An Alexa Fluor® 394-conjugated Donkey anti-rabbit IgG polyclonal was used as the secondary antibody at a dilution of 1/200.
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Immunocytochemistry/ Immunofluorescence - Anti-LC3B antibody (ab48394)ab48394 staining LC3B (LC3-II) in HeLa cells treated with calmidazolium chloride (ab120658), by ICC/IF. Increase of LC3B (LC3-II) expression correlates with increased concentration of calmidazolium chloride, as described in literature.
The cells were incubated at 37°C for 6h in media containing different concentrations of ab120658 (calmidazolium chloride) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab48394 (1 μg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LC3B antibody (ab48394)Immunohistochemistry analysis of (formalin-fixed,paraffin-embedded sections) analysis of human brain,cerebral cortex,neurons with cell processes LC3B with ab48394
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Immunocytochemistry/ Immunofluorescence - Anti-LC3B antibody (ab48394)Immunocytochemistry/Immunofluorescence analysis of HeLa cells with Dylight 488 (green).Nuclei and alpha -tubulin were counterstained with DAPI (blue) and Dylight 550 (red).
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Western blot - Anti-LC3B antibody (ab48394)This image is courtesy of an anonymous AbreviewAll lanes : Anti-LC3B antibody (ab48394) at 1/2000 dilution
Lane 1 : Rat whole tissue lysate - Normal liver
Lane 2 : Rat whole tissue lysate - liver treated with AEE788 at 50 mg/kg 3 times a week for 1 week
Lane 3 : Rat whole tissue lysate - liver treated with RAD at 2.5 mg/kg daily for 1 week
Lysates/proteins at 30 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/5000 dilution
Developed using the ECL technique.
Performed under non-reducing conditions.
Predicted band size: 15 kDa
Observed band size: 17 kDa (why is the actual band size different from the predicted?)
Exposure time: 1 minute
Protocols
References
This product has been referenced in:
- Magalhaes J et al. Effects of ambroxol on the autophagy-lysosome pathway and mitochondria in primary cortical neurons. Sci Rep 8:1385 (2018). WB ; Mouse . Read more (PubMed: 29362387) »
- Paiva NM et al. Ileal pouch of ulcerative colitis and familial adenomatous polyposis patients exhibit modulation of autophagy markers. Sci Rep 8:2619 (2018). WB ; Human . Read more (PubMed: 29422639) »
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