Anti-LC3B antibody (ab51520)
Key features and details
- Rabbit polyclonal to LC3B
- Suitable for: IHC-Fr, Flow Cyt (Intra), WB, ICC/IF
- Reacts with: Human
- Isotype: IgG
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Overview
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Product name
Anti-LC3B antibody
See all LC3B primary antibodies -
Description
Rabbit polyclonal to LC3B -
Host species
Rabbit -
Specificity
ab51520 is specific for the B isoform of LC3. -
Tested applications
Suitable for: IHC-Fr, Flow Cyt (Intra), WB, ICC/IFmore details -
Species reactivity
Reacts with: 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
- ICC/IF: HeLa, U373-MGc cells. IHC-P: U87-MG cells. Flow Cyt (Intra): NTERA-2 cells. IHC-P-Fr: U373-MG cells.
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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.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: PBS, 1% BSA -
Concentration information loading... -
Purity
Immunogen affinity purified -
Primary antibody notes
ab51520 has been shown to work on human malignant brain tumors, breast cancer, leukemia samples, and murine cells. -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
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-Fr | (2) |
1/2000.
|
| Flow Cyt (Intra) |
1/200.
|
|
| WB | (9) |
1/3000. Predicted molecular weight: 15 kDa. Detects bands of ~16 kDa (LC3-II) and ~18 kDa (LC3-I).
|
| ICC/IF | (3) |
1/2000.
|
| Notes |
|---|
|
IHC-Fr
1/2000. |
|
Flow Cyt (Intra)
1/200. |
|
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
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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
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 localization
Cytoplasm > 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: 81631 Human
- Omim: 609604 Human
- SwissProt: Q9GZQ8 Human
- Unigene: 356061 Human
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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 (ab51520)All lanes : Anti-LC3B antibody (ab51520) at 1/3000 dilution
Lane 1 : Wild-type HepG2 untreated control cell lysate
Lane 2 : Wild-type HepG2 Treated Chloroquine (50 uM, 16 h) cell lysate
Lane 3 : MAP1LC3B knockout HepG2 untreated control cell lysate
Lane 4 : MAP1LC3B knockout HepG2 Treated Chloroquine (50 uM, 16 h) cell lysate
Lane 5 : U-87 MG cell lysate
Lane 6 : PC-12 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 15 kDa
Observed band size: 14,16 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-LC3B antibody staining at 1/3000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab51520 was shown to bind specifically to LC3B. A band was observed at 16/14 kDa (yellow arrows) in treated wild-type HepG2 cell lysates with no signal observed at this size in MAP1LC3B knockout cell line ab277828 (knockout cell lysate ab283796). To generate this image, wild-type and MAP1LC3B knockout HepG2 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
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Immunohistochemistry (Frozen sections) - Anti-LC3B antibody (ab51520)Ab51520 at 1/2000 dilution staining treated U87-MG cultured and subcutaneous tumors
1) Control
2) Temozolomide -
Western blot - Anti-LC3B antibody (ab51520)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 - Anti-LC3B antibody (ab51520)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).
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Flow Cytometry (Intracellular) - Anti-LC3B antibody (ab51520)Flow cytometry (Intracellular) analysis of NTERA-2 cells with ab51520 at 1/200 using Dylight-488 conjugated goat anti-rabbit IgG secondary.
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Immunocytochemistry/ Immunofluorescence - Anti-LC3B antibody (ab51520)Ab51520 at 1/2000 dilution staining treated U373-MG cells;
1) Control
2) C2-ceramide.
The nuclei were stained with DAPI. -
Immunohistochemistry (Frozen sections) - Anti-LC3B antibody (ab51520)Ab51520 at 1/2000 dilution staining treated U373-MG (human glioblastoma) cells
1) Control
2) Temozolomide -
Immunocytochemistry/ Immunofluorescence - Anti-LC3B antibody (ab51520)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).
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Immunocytochemistry/ Immunofluorescence - Anti-LC3B antibody (ab51520)Immunocytochemistry/Immunofluorescence analysis of HeLa cells using anti-LC3B antibody(red). Nuclei were counterstained with DAPI (blue).
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (399)
ab51520 has been referenced in 399 publications.
- Adamczyk-Grochala J et al. DNMT2/TRDMT1 gene knockout compromises doxorubicin-induced unfolded protein response and sensitizes cancer cells to ER stress-induced apoptosis. Apoptosis 28:166-185 (2023). PubMed: 36273376
- Peng F et al. AMPK/MFF Activation: Role in Mitochondrial Fission and Mitophagy in Dry Eye. Invest Ophthalmol Vis Sci 63:18 (2022). PubMed: 36374514
- Li W et al. Shikonin induces ferroptosis in multiple myeloma via GOT1-mediated ferritinophagy. Front Oncol 12:1025067 (2022). PubMed: 36387145
- Li W et al. Fasudil prevents neomycin-induced hair cell damage by inhibiting autophagy through the miR-489/NDP52 signaling pathway in HEI-OC1 cells. Exp Ther Med 23:43 (2022). PubMed: 34849158
- Ferreira CB et al. S327 phosphorylation of the presynaptic protein SEPTIN5 increases in the early stages of neurofibrillary pathology and alters the functionality of SEPTIN5. Neurobiol Dis 163:105603 (2022). PubMed: 34954322