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LC3 Antibody (APG8B) (N-term)
Purified Rabbit Polyclonal Antibody (Pab)

Catalog # Source: Lot # Size:
AP1802a Rabbit SH050530I 0.1 mg
Concentration: Accession: Clone Name: Isotype:
0.25 mg/ml NP_073729, Q9GZQ8 RB7481 Rabbit Ig
Applications: Reactivity: MW (kDa):
WB, IF, IHC, E H, M, R, B*, C*, Z* 14557 Da
Target/Specificity:
This antibody is generated from rabbits immunized with a KLH conjugated synthetic peptide at the N-terminal region of human LC3 (APG8b).
Other Names:
MAP1LC3B, LC3B; MAP1A/1BLC3; MAP1LC3B
Application Data:
LC3 Antibody (APG8B) (N-term)

Left: Western blot analysis of anti-LC3 (APG8b) Pab (Cat. #AP1802a) in rat brain lysate. Both non-lipidated (arrow, I) and lipidated LC3 (APG8b) (arrow, II) were detected in membrane fraction (P) but pro-LC3 (APG8b) and non-lipidated LC3 ((APG8b) were detected in soluble fraction (S). Right: Western blot analysis of anti-LC3 (APG8b) Pab (Cat. #AP1802a) in 293 cell line lysates transiently transfected with the LC3 (APG8b) gene (2ug/lane). Human LC3 (APG8b) (arrow) was detected using the purified Pab (1:60 dilution).

 LC3 Antibody (APG8B) (N-term)

TOP: FACS sorted LSK cells (Lineage-Sca-1+c-kit+, enriched for hematopoietic stem cells) incubated with LC3 antibody (Cat.# AP1802a) at 1:50 dilution at 4 degree overnight., and detected with AlexaFlour 488 conjugated anti-rabbit antibody (1:200). Data courtesy of Wenbin Xiao, La Jolla Institute for Allergy and Immunology. CENTER: Figure 1 - Immunofluorescence analysis of anti-LC3 (APG8b) polyclonal antibody in rat brain before (left) and after (right) treatment with rapamycin (100 nm for 4 hr). CENTER: Figure 2 - Formalin-fixed and paraffin-embedded human cancer tissue reacted with the primary antibody, which was peroxidase-conjugated to the secondary antibody, followed by AEC staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated. BC = breast carcinoma; HC = hepatocarcinoma. BOTTOM: Figure 1 - Wild-type (Cln3+/+) or homozygous Cln3Äex7/8 (Cln3Äex7/8/Äex7/8) paraffin-embedded brain sections immunostained for the LC3 protein (Cat. # AP1802a LC3 antibody). Shown are the CA2/CA3 region of hippocampus (Hc) and cerebellum (Cb) from 10-month-old mice. Few immunopositive puncta are present in wild-type sections, whereas homozygous Cln3Äex7/8 sections contain clusters of LC3-positive puncta around pyramidal neurons and Purkinje cells (P). MOL, molecular layer; GCL, granule cell layer. Data courtesy of Dr. Susan Cotman, Massachusets General Hospital. BOTTOM: Figure 2 - Immunoblots for LC3 protein (Cat. # AP1802a LC3 antibody) are shown for extracts from untreated or rapamycin-treated (250 nM, 4 h) cultures of wild-type (CbCln3+/+) or homozygous cerebellar cells (CbCln3Äex7/8/Äex7/8). Rapamycin increases the LC3-II/LC3-I ratio in wild-type and homogyzous cerebellar cell extracts (2.6 and 2.2-fold, respectively), consistent with activation of autophagy.
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. MAP1LC3b 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.
Background References:
  1. Baehrecke EH. Nat Rev Mol Cell Biol. 6(6):505-10. (2005)
  2. Lum JJ, et al. Nat Rev Mol Cell Biol. 6(6):439-48. (2005)
  3. Greenberg JT. Dev Cell. 8(6):799-801. (2005)
  4. Levine B. Cell. 120(2):159-62. (2005)
  5. Shintani T and Klionsky DJ. Science. 306(5698):990-5. (2004)
  6. Tanida I., et al. Int. J. Biochem. Cell Biol. 36:2503-2518(2004)
  7. He H., et al. J. Biol. Chem. 278:29278-29287(2003)
  8. Tanida I., et al. J. Biol. Chem. 279:36268-36276(2004)
Product Citations:
  1. Nicholas L. Cianciola, et al. J. Cell Biol. 2009; 187:537-552. (E-pub)
  2. Nicholas L. Cianciola, et al. J. Cell Biol. 2009; 187:537-552. (E-pub)
  3. Shin-ichiro Hayashi, et al. Am. J. Pathol. 2009; 175:2226-2234. (E-pub)
  4. Lena Wartosch, Jens C. Fuhrmann, Michaela Schweizer, Tobias Stauber, and Thomas J. Jentsch, et al. FASEB J. published 6 August 2009, 10.1096/fj.09-130880. (Applications: WB)(E-pub)
  5. Mona Bains, et al. J. Biol. Chem. 2009; 284:20398-20407. (Applications: WB)
  6. Wang YH, et al. Experimental Biology and Medicine, Feb 2009; 234: 171 - 180. (Applications: WB, IF)
  7. Jennifer M. Rosenbluth, et al. Mol. Cell. Biol., Oct 2008; 28: 5951 - 5964.
  8. Jean H. Overmeyer, et al. Mol. Cancer Res., Jun 2008; 6: 965 - 977.
  9. Liao, Guanghong, et al. Am. J. Pathol. 2007 Sep 01;171(3):962-975. (Applications: IF)
  10. Cao, Yi, et al. J. Biol. Chem. 2006 Jul 21;281(29):20483-20493. (Applications: WB, IF)
Application Notes:
The suggested dilution is:
ELISA1:1,000
Western blotting1:100~500
Immunohistochemistry1:50~100
Immunofluorescence1:50
Format:
Purified polyclonal antibody supplied in PBS with 0.09% (W/V) sodium azide. This antibody is prepared by Saturated Ammonium Sulfate (SAS) precipitation followed by dialysis against PBS.
Storage:
Maintain refrigerated at 2-8°C for up to 6 months. For long term storage store at -20°C.
Precautions:
This product is for research use only. Not for use in diagnostic or therapeutic procedures.