Overview

  • Product name
  • Description
    Rabbit polyclonal to LC3B
  • Tested applications
    Suitable for: ICC/IF, IP, IHC-P, WB, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Dog, Human, Zebrafish, Syrian hamster, Bacteria
    Predicted to work with: Cow
  • 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

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).

IHC-Fr Use at an assay dependent concentration.

Target

  • Function
    Probably involved in formation of autophagosomal vacuoles (autophagosomes).
  • Tissue specificity
    Most abundant in heart, brain, skeletal muscle and testis. Little expression observed in liver.
  • Sequence similarities
    Belongs to the MAP1 LC3 family.
  • Post-translational
    modifications
    The 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 localization
    Cytoplasm > cytoskeleton. Endomembrane system. Cytoplasmic vesicle > autophagosome membrane. LC3-II binds to the autophagic membranes.
  • Information by UniProt
  • Database links
  • Alternative names
    • ATG8F antibody
    • Autophagy-related protein LC3 B antibody
    • Autophagy-related ubiquitin-like modifier LC3 B antibody
    • LC3B antibody
    • LC3II antibody
    • MAP1 light chain 3 like protein 2 antibody
    • MAP1 light chain 3-like protein 2 antibody
    • MAP1A/1BLC3 antibody
    • MAP1A/MAP1B LC3 B antibody
    • MAP1A/MAP1B light chain 3 B antibody
    • MAP1ALC3 antibody
    • MAP1LC3B a antibody
    • Map1lc3b antibody
    • Microtubule associated protein 1 light chain 3 beta antibody
    • Microtubule-associated protein 1 light chain 3 beta antibody
    • Microtubule-associated proteins 1A/1B light chain 3B antibody
    • MLP3B_HUMAN antibody
    see all

Anti-LC3B antibody images

  • 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
    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.
  • 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.

    See Abreview

  • 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 analysis of (formalin-fixed,paraffin-embedded sections) analysis of human brain,cerebral cortex,neurons with cell processes LC3B with ab48394

  • Immunocytochemistry/Immunofluorescence analysis of HeLa cells with Dylight 488 (green).Nuclei and alpha -tubulin were counterstained with DAPI (blue) and Dylight 550 (red).

  • All 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
    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

    This image is courtesy of an anonymous Abreview

    See Abreview

References for Anti-LC3B antibody (ab48394)

This product has been referenced in:
  • Moretton A  et al. Selective mitochondrial DNA degradation following double-strand breaks. PLoS One 12:e0176795 (2017). WB . Read more (PubMed: 28453550) »
  • Vazquez-Cintron EJ  et al. Engineering Botulinum Neurotoxin C1 as a Molecular Vehicle for Intra-Neuronal Drug Delivery. Sci Rep 7:42923 (2017). Read more (PubMed: 28220863) »

See all 58 Publications for this product

Product Wall

Application
Western blot
Sample
Human Cell lysate - whole cell (LNCaP - prostate cancer cells)
Gel Running Conditions
Reduced Denaturing (15% SDS-PAGE)
Loading amount
25 µg
Specification
LNCaP - prostate cancer cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

Submitted Jun 03 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (LNCaP - prostate cancer cells)
Permeabilization
Yes - 0.2% Triton X-100
Specification
LNCaP - prostate cancer cells
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
Fixative
Formaldehyde
Username

Dr. Armen Petrosyan

Verified customer

Submitted Jun 03 2016

Application
Western blot
Sample
Rat Cell lysate - whole cell (Rat Hepatocytes)
Gel Running Conditions
Reduced Denaturing (15% SDS-PAGE)
Loading amount
20 µg
Specification
Rat Hepatocytes
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

Submitted Jun 03 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (Rat hepatocytes)
Permeabilization
Yes - 0.2% Triton X-100
Specification
Rat hepatocytes
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
Fixative
Formaldehyde
Username

Dr. Armen Petrosyan

Verified customer

Submitted Mar 26 2016

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Pancreatic cancer xenograft)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH 9
Permeabilization
No
Specification
Pancreatic cancer xenograft
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 15% · Temperature: 20°C
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Dec 18 2015

Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Reduced Denaturing
Sample
Human Cell lysate - whole cell (HEK293)
Specification
HEK293
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Feb 06 2015

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Sample
Mouse Cell (mouse heart myocyte)
Specification
mouse heart myocyte
Permeabilization
Yes - 1% TRITON-X-100
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Feb 04 2015

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Sample
Rat Cell (h9c2)
Specification
h9c2
Permeabilization
Yes - 1% TRITON-X-100
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Feb 04 2015

Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Reduced Denaturing
Sample
Chinese Hamster Cell lysate - whole cell (CHO cell)
Specification
CHO cell
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Feb 03 2015

Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Reduced Denaturing
Sample
Rat Cell lysate - whole cell (h9c2)
Specification
h9c2
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Feb 03 2015

1-10 of 49 Abreviews or Q&A

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