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RecombinantRabMAb

Recombinant Anti-MAP1LC3A antibody [EP1983Y] - Autophagosome Marker (ab52768)

Submit a review Q&A (2)References (13)
Immunocytochemistry/ Immunofluorescence - Anti-MAP1LC3A antibody [EP1983Y] - Autophagosome Marker (ab52768)
  • Flow Cytometry (Intracellular) - Anti-MAP1LC3A antibody [EP1983Y] - Autophagosome Marker (ab52768)
  • Western blot - Anti-MAP1LC3A antibody [EP1983Y] - Autophagosome Marker (ab52768)
  • Immunocytochemistry/ Immunofluorescence - Anti-MAP1LC3A antibody [EP1983Y] - Autophagosome Marker (ab52768)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MAP1LC3A antibody [EP1983Y] - Autophagosome Marker (ab52768)
  • Anti-MAP1LC3A antibody [EP1983Y] - Autophagosome Marker (ab52768)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP1983Y] to MAP1LC3A - Autophagosome Marker
  • Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
  • Reacts with: Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Carrier Free PE

Overview

  • Product name

    Anti-MAP1LC3A antibody [EP1983Y] - Autophagosome Marker
    See all MAP1LC3A primary antibodies

  • Description

    Rabbit monoclonal [EP1983Y] to MAP1LC3A - Autophagosome Marker

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details

  • Species reactivity

    Reacts with: Rat, Human
    Predicted to work with: Mouse

  • Immunogen

    Synthetic peptide within Human MAP1LC3A aa 100 to the C-terminus (C terminal). The exact sequence is proprietary.
    Database link: Q9H492

  • Positive control

    • WB: Hela cell lysate IHC-P: Human breast carcinoma tissue

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab52768 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
Flow Cyt (Intra)
1/40 - 1/1000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
WB
1/5000 - 1/10000. Detects a band of approximately 16 kDa (predicted molecular weight: 14 kDa).
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
1/100 - 1/500.
Notes
Flow Cyt (Intra)
1/40 - 1/1000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
WB
1/5000 - 1/10000. Detects a band of approximately 16 kDa (predicted molecular weight: 14 kDa).
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
1/100 - 1/500.

Target

  • Function

    Probably involved in formation of autophagosomal vacuoles (autophagosomes).

  • Tissue specificity

    Most abundant in heart, brain, liver, skeletal muscle and testis but absent in thymus and peripheral blood leukocytes.

  • Sequence similarities

    Belongs to the MAP1 LC3 family.

  • Post-translational
    modifications

    The precursor molecule is cleaved by APG4B/ATG4B to form the cytosolic form, LC3-I. This is activated by APG7L/ATG7, transferred to ATG3 and conjugated to phospholipid to form the membrane-bound 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

    see all

  • Alternative names

    • ATG8E antibody
    • Autophagy related protein LC3 A antibody
    • Autophagy related ubiquitin like modifier LC3 A antibody
    • Autophagy-related protein LC3 A antibody
    • Autophagy-related ubiquitin-like modifier LC3 A antibody
    • LC3 antibody
    • LC3A antibody
    • MAP1 light chain 3 like protein 1 antibody
    • MAP1 light chain 3-like protein 1 antibody
    • MAP1A/1B light chain 3 A antibody
    • MAP1A/MAP1B LC3 A antibody
    • MAP1A/MAP1B light chain 3 A antibody
    • MAP1ALC3 antibody
    • MAP1BLC3 antibody
    • Map1lc3a antibody
    • Microtubule associated proteins 1A/1B light chain 3 antibody
    • Microtubule-associated protein 1 light chain 3 alpha antibody
    • Microtubule-associated proteins 1A and 1B, light chain 3 antibody
    • Microtubule-associated proteins 1A/1B light chain 3A antibody
    • MLP3A_HUMAN antibody
    see all

Images

  • Immunocytochemistry/ Immunofluorescence - Anti-MAP1LC3A antibody [EP1983Y] - Autophagosome Marker (ab52768)
    Immunocytochemistry/ Immunofluorescence - Anti-MAP1LC3A antibody [EP1983Y] - Autophagosome Marker (ab52768)

    Immunocytochemistry/Immunofluorescence analysis of C6 (rat glioma) labelling MAP1LC3A with purified ab52768 at 1/500. Cells were fixed with 100% methanol. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (Ab150077). Nuclei counterstained with DAPI (blue).

  • Flow Cytometry (Intracellular) - Anti-MAP1LC3A antibody [EP1983Y] - Autophagosome Marker (ab52768)
    Flow Cytometry (Intracellular) - Anti-MAP1LC3A antibody [EP1983Y] - Autophagosome Marker (ab52768)

    Overlay histogram showing SHSY-5Y cells stained with ab52768 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab52768, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in SHSY-5Y cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

     

  • Western blot - Anti-MAP1LC3A antibody [EP1983Y] - Autophagosome Marker (ab52768)
    Western blot - Anti-MAP1LC3A antibody [EP1983Y] - Autophagosome Marker (ab52768)
    Anti-MAP1LC3A antibody [EP1983Y] - Autophagosome Marker (ab52768) at 1/10000 dilution + Hela cell lysate at 10 µg

    Secondary
    Goat anti-rabbit HRP labeled at 1/2000 dilution

    Predicted band size: 14 kDa
    Observed band size: 16 kDa why is the actual band size different from the predicted?

  • Immunocytochemistry/ Immunofluorescence - Anti-MAP1LC3A antibody [EP1983Y] - Autophagosome Marker (ab52768)
    Immunocytochemistry/ Immunofluorescence - Anti-MAP1LC3A antibody [EP1983Y] - Autophagosome Marker (ab52768)Image from Buckingham EM et al., Biotechniques. 2014;57(5):241-4. Fig 1.; doi: 10.2144/000114226.

    Immunocytochemistry/Immunofluorescence analysis of human keratinocytes (pannel A) and VZV-infected melanoma cells (panel E) labelling MAP1LC3A with ab52768.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MAP1LC3A antibody [EP1983Y] - Autophagosome Marker (ab52768)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MAP1LC3A antibody [EP1983Y] - Autophagosome Marker (ab52768)

    Immunohistochemical staining of MAP1LC3A in paraffin embedded human breast carcinoma using ab52768 at a 1/100 dilution.

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Anti-MAP1LC3A antibody [EP1983Y] - Autophagosome Marker (ab52768)
    Anti-MAP1LC3A antibody [EP1983Y] - Autophagosome Marker (ab52768)

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

References (13)

Publishing research using ab52768? Please let us know so that we can cite the reference in this datasheet.

ab52768 has been referenced in 13 publications.

  • Mori T  et al. Autophagy is involved in the sclerotic phase of systemic sclerosis. Fukushima J Med Sci 66:17-24 (2020). PubMed: 32281584
  • Gao J  et al. Autophagy activation promotes clearance of a-synuclein inclusions in fibril-seeded human neural cells. J Biol Chem 294:14241-14256 (2019). PubMed: 31375560
  • Hirai Y  et al. Autophagy in malnutrition-associated dermatoses. J Dermatol 46:43-47 (2019). PubMed: 30379352
  • Xu Y  et al. Protective Effects of Melatonin Against Zearalenone Toxicity on Porcine Embryos in vitro. Front Pharmacol 10:327 (2019). PubMed: 31024301
  • Horigome Y  et al. Loss of autophagy in chondrocytes causes severe growth retardation. Autophagy N/A:1-11 (2019). PubMed: 31203752
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

1-2 of 2 Abreviews or Q&A

Question

I have another question regarding the anti-MAP1LC3a Ab (ab52768). 1) Based on the Western blot data shown in the web page, I am assuming that this Ab binds only to the uncleaved form of LC3? 2) Have you tested it for the cleaved form of LC3 as well? 3) Where does this antibody bind toon LC3? 4) Do you have Abs that bind only to the uncleaved and only to the cleaved forms of LC3?

Read More
Answer

Thank you for your patience. I have heard back from the lab with the following information: 1) This antibody binds to the uncleaved and cleaved form of LC3, LC3-I. 2) It should recognize two isoforms, LC3-I and LC3-II. One of our scientists recommended this product for detecting both isoforms. The 16-kDa (LC3-II) and 18-kDa bands (LC3-I) are probably close together on the gel, so the band width is unusually large. 3) The peptide maps near the C-terminal end of human MAP1LC3A (UniProt: Q9H492). The immunogen does not contain aa 121. We believe the cleavage site refers to the 120 site according to the literature. 4) Since the cleavage site is 120 and the difference between cleaved and uncleaved form is 1 amino acid, it would be in general very difficult to raise an antibody that differenciates between both forms. So, we do not have such an antibody available, unfortunately. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Question

I have another question regarding the anti-MAP1LC3a Ab (ab52768). Based on the Western blot data shown in the web page, I am assuming that this Ab binds only to the uncleaved form of LC3? Have you tested it for the cleaved form of LC3 as well? Where does this antibody bind toon LC3? Do you have Abs that bind only to the uncleaved and only to the cleaved forms of LC3?

Read More
Answer

Thank you for your email. Yes, it went directly into my inbox. Thank you for your kind words. To answer your questions: I have checked the database entry for the human protein: http://www.uniprot.org/uniprot/Q9H492 and am not able to find information about the cleavage site. Unless you are refering to the removal of residue 121? Please let me know. Once I have this information Iwill also be able to check for antiibodies against the cleaved und uncleaved form of LC3. The antibody was raised against the C-terminal region, but I will have to check with the lab about more detailed information regarding the immunogen region. I will be in touch as soon as I hear back from the lab. In the meantime, please let me know the cleavage site as well. Thank you, and I look forward to hear back from you.

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