Autophagy 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).

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.