LC3B (MAP1LC3B) (N-term) (incl. pos. control) Mouse Monoclonal Antibody [Clone ID: 2G6]

CAT#: AM20213PU-N

LC3B (MAP1LC3B) (N-term) (incl. pos. control) mouse monoclonal antibody, clone 2G6, Purified

Product Images

Detection of endogenous LC-3: Whole cell lysates of untreated tumor cells were applied to SDS-PAGE and transferred to a PVDF membrane. The immunoblot was probed with LC3 antibody (Clone: 2G6) at 0.5 ug/ ml for 1h at RT and developed by ECL (exp. time: 30 sec). Lane 1: HeLa Lane 2: HepG2 Lane 3: HEK 293 Lane 4: SH-SY5Y Lane 5: MDCK Lane 6: PC12 Lane 7: CMT Lane 8: Neuro2A Lane 9: NIH-3T3

Specifications

Product Data

• Clone Name: 2G6
• Applications: IF, WB
• Recommended Dilution: Immunoblotting: 0.5 µg/ml for HRPO/ECL detection
  Recommended blocking buffer: Casein/Tween 20 based blocking and blot incubation buffer.
  We strongly recommend to use PVDF membranes for immunoblot analysis.
  Immunocytochemistry: Use at 1-10 µg/ml (Paraformaldehyd/Methanol fixation).
  Included Positive Control: Cell lysate from untreated Neuro 2A (See Protocol below).
• Reactivities: Hamster, Human, Monkey, Mouse, Rat
• Host: Mouse
• Isotype: IgG1
• Clonality: Monoclonal
• Immunogen: Synthetic peptide hemocyanin conjugated derived from the N-terminus of LC3-B
• Specificity: This antibody specifically recognizes both forms of endogenous LC3, the cytoplasmic LC3-I (18 kDa) as well as the lipidated form generated during autophagosome and autophagolysosome formation: LC3-II (16 kDa).
• Formulation: PBS containing 0.09% Sodium Azide, PEG and Sucrose/50% Glycerol
  State: Purified
  State: Liquid purified IgG fraction
• Concentration: 0.1 mg/ml
• Purification: Subsequent Ultrafiltration and Size Exclusion Chromatography
• Gene Name: Homo sapiens microtubule associated protein 1 light chain 3 beta (MAP1LC3B)
• Database Link:
  Entrez Gene 64862 RatEntrez Gene 67443 MouseEntrez Gene 81631 Human
• Background: Autophagy is an alternative process of proteasomal degradation for some long-lived proteins or organelles. Alterations in the autophagic-lysosomal compartment have been linked to neuronal death in many neurodegenerative disorders as well as in transmissible neuronal pathologies (prion diseases). Genetic studies in yeast have shown that Autophagy-defective Gene-8 (Atg-8) represents a specific marker for autophagy. Among the four families of mammalian Atg8-related proteins only LC3 (microtubule-associated protein1 light chain 3) is expressed at sufficient high levels and efficiently recruited to autophagic vesicles in cells and tissues. During autophagy the cytoplasmic form, LC3-I is processed and recruited to autophagosomes, where LC3-II is generated by site specific proteolysis near to the C-terminus. Autophagic vacuoles have been also reported frequently in cardiomyopathies or muscle cells exposed to different experimental settings.
• Synonyms: MAP1LC3B, MAP1A/MAP1B, Map1lc3b, Map1alc3, Map1lc3
• Note: Molecular Weight: 18 kDa (LC3-I), 16 kDa (LC-II)
  
  Protocol: Positive Control: Cell lysate from untreated Neuro 2A cells, brain endothelioma (Mouse)
  
  Format: Lyophilized cell lysate from serum starved Neuro 2A.
  
  Reconstitution: Restore by addition of 200 µl H2O. After complete solubilization add 200 µl 2x SDS-PAGE sample buffer, mix and incubate at 90°C for 5 min.
  
  Application: The positive control cell lysate is recommended for immunoblot applications. 20 µl of positive control cell lysate correspond to ca. 20.000 cells.
  Use 20 µl/lane (mini gel) for HRPO/ECL detection of the target proteins.
  Please NOTE: The lyophilized cell lysates conatin SDS and are not recommended for applications with native proteins such as in immunoprecipitation.
  
  Storage: Aliquote reconstituted product and store frozen. Avoid repeated fereezing and thawing.