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

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

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 115 Publications for this product

Customer reviews and Q&As


We recommend a heat-mediated retrieval, with 10mM citrate buffer, pH 6.0. Heat the buffer to boiling, place the slides in the buffer (after de-paraffinization) and maintain the temperature at 95-100C for 20 minutes. After 20 minutes, allow the buf...

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I am sorry to hear that you have been experiencing problems using this product in the application that you wish.

In order to assess the quality of our products I would ask that you complete a brief questionnaire relating to the application us...

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I am very sorry to hear that you have been experiencing issues with ab48394.

I would be very happy to provide a free of charge replacement for you provided that this product has been used according to the the terms of our 100% Abpromise guar...

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Thank you for contacting Abcam. I do wish to apologize for the delay in getting this information to you.

We have been in touch with the lab regarding your question of specificity of ab48394 to LC3B. As the immunogen sequence has been conside...

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Unfortunately, we didn't test this antibody for which form of LC3B it is specific.


However, according to the literature it looks like the two types are derived via carboxyterminal cleaving. As the antibody is derived from the N- terminal ...

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Thank you for your reply.

Our LC3 antibody, ab48394, has been raised using a synthetic peptide made to N-terminal portion of the human LC3 protein sequence (between residues 1-100) and in our validation studies, it has shown high affinity ...

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Thank you for contacting us.

AB48394 was created from a synthetic peptide immunogen corresponding to the N terminus regionof the human LC3 protein. We do believe this product to be specific to this protein. If you have been experiencing is...

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Thank you for your message and for providing this further information.

I am sorry to hear the suggestions made have not improved the results on this occasion. I appreciate the time you have spent on these experiments and would be pleased to ...

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Thank you for taking the time to contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from the latest batch of this antibody, GR63250-1 .

I would like to reassure you that this antibody is tested and covered ...

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Thank you for taking time to complete our questionnaire and for contacting us. I am sorry to hear this antibody is not providing satisfactory results.

The details provided will enable us to investigate this case and will provide us with vita...

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1-10 of 21 Q&A

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