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LC3 Antibody (APG8A) (P45)

Peptide Affinity Purified Rabbit Polyclonal Antibody (Pab)

     
  • IF - LC3 Antibody (APG8A) (P45) AP1801b
    Fluorescent image of U251 cells stained with AP1801b LC3 (APG8A) (P45) antibody. U251 cells were treated with Chloroquine (50 μM,16h), then fixed with 4% PFA (20 min), permeabilized with Triton X-100 (0.2%, 30 min). Cells were then incubated with AP1801b LC3 (APG8A) (P45) primary antibody (1:100, 2 h at room temperature). For secondary antibody, Alexa Fluor® 488 conjugated donkey anti-rabbit antibody (green) was used (1:1000, 1h). Nuclei were counterstained with Hoechst 33342 (blue) (10 μg/ml, 5 min). LC3 immunoreactivity is localized to autophagic vacuoles in the cytoplasm of U251 cells.
  • WB - LC3 Antibody (APG8A) (P45) AP1801b
    Western blot analysis of LC3 (APG8a) (arrow) using purified Pab (Cat.#AP1801b). 293 cell lysates (2 ug/lane) either nontransfected (Lane 1) or transiently transfected with the LC3 (APG8a) gene (Lane 2) (Origene Technologies).
  • IHC-P - LC3 Antibody (APG8A) (P45) AP1801b
    Formalin-fixed and paraffin-embedded heart muscle tissue reacted with Autophagy LC3 APG8a (MAP1LC3A) Antibody (P45) (Cat.#AP1801b), which was peroxidase-conjugated to the secondary antibody, followed by DAB staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated.
  • SPECIFICATION
  • CITATIONS: 2
  • PROTOCOLS
  • BACKGROUND
Product Information
Application
  • Applications Legend:
  • WB=Western Blot
  • IHC=Immunohistochemistry
  • IHC-P=Immunohistochemistry (Paraffin-embedded Sections)
  • IHC-F=Immunohistochemistry (Frozen Sections)
  • IF=Immunofluorescence
  • FC=Flow Cytopmetry
  • IC=Immunochemistry
  • ICC=Immunocytochemistry
  • E=ELISA
  • IP=Immunoprecipitation
  • DB=Dot Blot
  • CHIP=Chromatin Immunoprecipitation
  • FA=Fluorescence Assay
  • IEM=Immunoelectronmicroscopy
  • EIA=Enzyme Immunoassay
WB, IHC-P, IF, E
Primary Accession Q9H492, Q9GZQ8
Other Accession A6NCE7, Q62625, Q9CQV6, O41515, Q6XVN8, Q91VR7, Q2HJ23, Q6NX90, Q9H492, Q9GZQ8
Reactivity Human
Predicted Zebrafish, Bovine, Mouse, Rat
Host Rabbit
Clonality Polyclonal
Isotype Rabbit Ig
Antigen Region 30-56 aa
Additional Information
Other Names Microtubule-associated proteins 1A/1B light chain 3A, Autophagy-related protein LC3 A, Autophagy-related ubiquitin-like modifier LC3 A, MAP1 light chain 3-like protein 1, MAP1A/MAP1B light chain 3 A, MAP1A/MAP1B LC3 A, Microtubule-associated protein 1 light chain 3 alpha, MAP1LC3A
Target/Specificity This LC3 antibody is generated from rabbits immunized with a KLH conjugated synthetic peptide between 30-56 amino acids from human LC3.
Dilution IF~~1:100
WB~~1:1000
IHC-P~~1:10~50
Format Purified polyclonal antibody supplied in PBS with 0.09% (W/V) sodium azide. This antibody is purified through a protein A column, followed by peptide affinity purification.
StorageMaintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
PrecautionsLC3 Antibody (APG8A) (P45) is for research use only and not for use in diagnostic or therapeutic procedures.
Protein Information
Research Areas
Citations ( 0 )

Background

Macroautophagy is the major inducible pathway for the general turnover of cytoplasmic constituents in eukaryotic cells, it is also responsible for the degradation of active cytoplasmic enzymes and organelles during nutrient starvation. Macroautophagy involves the formation of double-membrane bound autophagosomes which enclose the cytoplasmic constituent targeted for degradation in a membrane bound structure, which then fuse with the lysosome (or vacuole) releasing a single-membrane bound autophagic bodies which are then degraded within the lysosome (or vacuole). MAP1A and MAP1B are microtubule-associated proteins which mediate the physical interactions between microtubules and components of the cytoskeleton. These proteins are involved in formation of autophagosomal vacuoles (autophagosomes). MAP1A and MAP1B each consist of a heavy chain subunit and multiple light chain subunits. MAP1LC3a is one of the light chain subunits and can associate with either MAP1A or MAP1B. 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.

References

References for protein:
1.Tanida, I., et al., J. Biol. Chem. 279(35):36268-36276 (2004).
2.He, H., et al., J. Biol. Chem. 278(31):29278-29287 (2003).
3.Mann, S.S., et al., J. Neurosci. Res. 43(5):535-544 (1996).

References for U251 cell line:
1. Westermark B.; Pontén J.; Hugosson R. (1973).” Determinants for the establishment of permanent tissue culture lines from human gliomas”. Acta Pathol Microbiol Scand A. 81:791-805. [PMID: 4359449].
2. Pontén, J.,Westermark B. (1978).” Properties of Human Malignant Glioma Cells in Vitro”. Medical Biology 56: 184-193.[PMID: 359950].
3. Geng Y.;Kohli L.; Klocke B.J.; Roth K.A.(2010). “Chloroquine-induced autophagic vacuole accumulation and cell death in glioma cells is p53 independent”. Neuro Oncol. 12(5): 473–481.[ PMID: 20406898].

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$ 295.00
$ 99.00
Cat# AP1801b
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(40 western blots)
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