Overview

  • Product name
  • Description
    Rabbit polyclonal to LC3B
  • Host species
    Rabbit
  • Specificity
    ab51520 is specific for the B isoform of LC3.
  • Tested applications
    Suitable for: IHC - Wholemount, Flow Cyt, IHC-Fr, WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide corresponding to Human LC3B (N terminal). Synthetic peptide made to the N terminal region of human LC3, isoform B protein.
    Database link: Q9GZQ8

  • Positive control
    • WB: Treated U87-MG lysates. ICC/IF: Treated U87-MG cells. Human malignant brain tumors, breast cancer, leukemia samples and murine cells
  • General notes

    ab51520 has been shown to work on human malignant brain tumors, breast cancer, leukemia samples, and murine cells.

    The product formulation changed to include 1% BSA on 17th May 2017. The following lot does not contain BSA and is still in stock GR320876. All future lots will contain 1% BSA.

Properties

Applications

Our Abpromise guarantee covers the use of ab51520 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
IHC - Wholemount Use at an assay dependent concentration.
Flow Cyt 1/200.
IHC-Fr 1/2000.
WB 1/3000. Predicted molecular weight: 15 kDa. Detects bands of ~16 kDa (LC3-II) and ~18 kDa (LC3-I).
ICC/IF 1/2000.

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

Images

  • All lanes : Anti-LC3B antibody (ab51520) at 1/3000 dilution

    Lane 1 : treated U87-MG (human glioblastoma astrocytoma) lysates with Arsenic Trioxide at 0 µM
    Lane 2 : treated U87-MG (human glioblastoma astrocytoma) lysates with Arsenic Trioxide at 2 µM
    Lane 3 : treated U87-MG (human glioblastoma astrocytoma) lysates with Arsenic Trioxide at 4 µM
    Lane 4 : treated U87-MG (human glioblastoma astrocytoma) lysates with Arsenic Trioxide at 8 µM

    Predicted band size: 15 kDa

  • Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling LC3B with ab51520.An Alexa Fluor 488-conjugated Goat to rabbit IgG was used as secondary antibody (green). Actin filaments were labeled with Alexa Fluor 568 phalloidin (red). DAPI was used to stain the cell nuclei (blue).

  • Ab51520 at 1/2000 dilution staining treated U87-MG cultured and subcutaneous tumors
    1) Control
    2) Temozolomide
  • Flow cytometry analysis of NTERA-2 cells with ab51520 at 1/200 using Dylight-488 conjugated goat anti-rabbit IgG secondary.

  • Ab51520 at 1/2000 dilution staining treated U373-MG cells;
    1) Control
    2) C2-ceramide.
    The nuclei were stained with DAPI.
  • Ab51520 at 1/2000 dilution staining treated U373-MG (human glioblastoma) cells
    1) Control
    2) Temozolomide
  • Immunocytochemistry/Immunofluorescence analysis of HeLa cells using anti-LC3B antibody(red) at 0.1 ug/ml. Nuclei were counterstained with DAPI (blue).Nuclei and alpha-tubulin were counterstained with DAPI (blue) and Dylight 550 (red).

  • Ab51520 at 1/2000 dilution staining gliobastoma multiform tissue
    1) normal brain
    2) gliobastoma multiform tissue
  • Immunocytochemistry/Immunofluorescence analysis of HeLa cells using anti-LC3B antibody(red). Nuclei were counterstained with DAPI (blue).

References

This product has been referenced in:
  • Zhang J  et al. Endothelial Monocyte-Activating Polypeptide-II Induces BNIP3-Mediated Mitophagy to Enhance Temozolomide Cytotoxicity of Glioma Stem Cells via Down-Regulating MiR-24-3p. Front Mol Neurosci 11:92 (2018). Read more (PubMed: 29632473) »
  • Ladds MJGW  et al. Autophagic flux blockage by accumulation of weakly basic tenovins leads to elimination of B-Raf mutant tumour cells that survive vemurafenib. PLoS One 13:e0195956 (2018). Read more (PubMed: 29684045) »
See all 80 Publications for this product

Customer reviews and Q&As

1-10 of 24 Abreviews or Q&A

Answer

Thank you for your reply.

I will be pleased to provide an free of charge vial of from a different lot.

Regrettably, this antibody is not in stock at the moment. Another batch is in production and is expected to be available by the end of June. I sincerely apologise for the inconvenience.

I can place the order for you, and it will be delivered to you as soon as it is available. Otherwise, you may like to consider selecting an altnerativeantibody, and I can provide a free of charge vial.

For example, the following LC3B antibodies have been tested in rat and WB:
ab63817 Anti-LC3B antibody
Rabbit polyclonal
Tested in: IHC-P, WB, ICC/IF
Reacts with: Human, Mouse, Rat
https://www.abcam.com/index.html?datasheet=63817 (or use the following: https://www.abcam.com/index.html?datasheet=63817).

ab64781 Anti-LC3B antibody
Rabbit polyclonal
Tested in: IHC-P, WB, IHC-Fr
Reacts with: Mouse, Rat
https://www.abcam.com/index.html?datasheet=64781 (or use the following: https://www.abcam.com/index.html?datasheet=64781).

Alternatively,I would be pleased to provide you with a credit note.

I look forward to hearing from you with details of how you would like to proceed.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Rat Tissue lysate - whole (primary hippocampal neuron)
Gel Running Conditions
Reduced Denaturing (gel 5%)
Loading amount
1 µg
Treatment
different concentration toxicant for 24hrs
Specification
primary hippocampal neuron
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C

Abcam user community

Verified customer

Submitted Jan 24 2018

Application
Western blot
Sample
Monkey Tissue lysate - whole (Cardiac)
Gel Running Conditions
Reduced Denaturing
Loading amount
25 µg
Specification
Cardiac
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Nov 01 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Cardic stem celll)
Permeabilization
Yes - Triton x-100, 0.01%
Specification
Cardic stem celll
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Dec 16 2016

Application
Immunocytochemistry
Sample
Mouse Cultured Cells (Cardiomyocytes)
Permeabilization
Yes - Triton x-100, 0.01%
Specification
Cardiomyocytes
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 4°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Dec 03 2015

Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (Heart)
Permeabilization
Yes - Triton x-100, 0.01%
Specification
Heart
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Nov 06 2015

Application
Flow Cytometry
Sample
Mouse Cell (Cardiomyocytes)
Permeabilization
Yes - Triton X-100, 0.01%
Gating Strategy
LC3B-AlexaFluor488 positive
Specification
Cardiomyocytes
Preparation
Cell harvesting/tissue preparation method: Trypsinization
Sample buffer: 0.25% for 30 sec
Fixation
Paraformaldehyde

Abcam user community

Verified customer

Submitted Sep 30 2015

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

Abcam user community

Verified customer

Submitted Sep 30 2015

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

Abcam user community

Verified customer

Submitted Sep 28 2015

Application
Immunocytochemistry
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1%
Sample
Human Cultured Cells (HEK293)
Specification
HEK293
Permeabilization
Yes - Triton x-100, 0.01%
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Jan 16 2015

1-10 of 24 Abreviews or Q&A

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