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20258
LAPosome Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

LAPosome Antibody Sampler Kit #20258

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LAPosome Antibody Sampler Kit: Image 1
Western blot analysis of extracts from various cell lines using UVRAG (D2Q1Z) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). HCT 116 cells have been reported to contain a monoallelic frameshift mutation resulting in significantly reduced levels of endogenous UVRAG expression (11).
LAPosome Antibody Sampler Kit: Image 2
Western blot analysis of extracts from various cell lines using PIK3R4 Antibody.
LAPosome Antibody Sampler Kit: Image 3
Western blot analysis of extracts from various cell lines using Beclin-1 (D40C5) Rabbit mAb.
LAPosome Antibody Sampler Kit: Image 4
Western blot analysis of extracts from mouse brain and C6 cells using PI3 Kinase Class III (D9A5) Rabbit mAb.
LAPosome Antibody Sampler Kit: Image 5
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
LAPosome Antibody Sampler Kit: Image 6
Western blot analysis of extracts from various cell lines using Rubicon (D9F7) Rabbit mAb.
LAPosome Antibody Sampler Kit: Image 7
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing human UVRAG (hUVRAG; +), using UVRAG (D2Q1Z) Rabbit mAb.
LAPosome Antibody Sampler Kit: Image 8
Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® Beclin-1 siRNA I #6222 (+) or SignalSilence® Beclin-1 siRNA II (+), using Beclin-1 (D40C5) XP® Rabbit mAb #3495 (upper) or α-Tubulin (11H10) Rabbit mAb #2125 (lower). The Beclin-1 (D40C5) XP® Rabbit mAb confirms silencing of Beclin-1 expression, while the α-Tubulin (11H10) Rabbit mAb is used to control for loading and specificity of Beclin-1 siRNA.
LAPosome Antibody Sampler Kit: Image 9
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with human Rubicon (hRubicon, +), using Rubicon (D9F7) Rabbit mAb. Rubicon construct was kindly provided by Dr. Qing Zhong, University of California, Berkeley, CA.
LAPosome Antibody Sampler Kit: Image 10
Immunoprecipitation of UVRAG from PANC-1 cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or UVRAG (D2Q1Z) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using UVRAG (D2Q1Z) Rabbit mAb.
To Purchase # 20258
Cat. # Size Qty. Price
20258T
1 Kit  (6 x 20 microliters) $ 402

Product Description

The LAPosome Antibody Sampler Kit provides an economical means of analyzing the proteins in the LAPosome complex. The kit includes enough antibodies to perform two western blot experiments with each primary antibody.

Specificity / Sensitivity

Each antibody in the LAPosome Antibody Sampler Kit detects endogenous levels of its target protein. Rubicon (D9F7) Rabbit mAb detects a band of unknown origin at 55 kDa.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Thr72 of human Beclin-1, Leu210 of human Rubicon, Lys630 of PI3 kinase class III, and Gly502 of human UVRAG. Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly825 of human PIK3R4 protein. Antibodies are purified by protein A and peptide affinity chromatography.

Background

LC3-associated phagocytosis (LAP) is a process at the crossroads between autophagy and phagocytosis in which uptake of extracellular particles into phagophores utilizes components of the autophagic machinery and is targeted to the lysosome for degradation (reviewed in 1-3). Activation of LAP by infectious agents through engagement of pathogen receptors like Toll-like receptors (TLRs) are important in immune regulation and host defense. While autophagosomes and LAP phagophores both express lipidated LC3, autophagosomes are double membrane vesicles while the phagophores in LAP are single membrane. While LAP shares some components of canonical autophagy, there are some distinct differences. Both autophagy and LAP require PI3 Kinase Class III (PI3KC3; also known as VPS34) as well as the regulatory protein PIK3R4 (also known as VPS15) and Beclin-1. However, while canonical autophagy requires Atg14 and Ambra-1 in this complex, LAP requires Rubicon and UVRAG. This complex, known as the LAPosome, activates PI3KC3 and triggers nicotinamide adenine dinucleotide phosphate (NADPH) oxidase 2 (NOX2), generating reactive oxygen species (ROS) for phagophore maturation. The expression of Rubicon in this complex has emerged as a defining characteristic of LAP.
  1. Heckmann, B.L. and Green, D.R. (2019) J Cell Sci 132, jcs222984. doi: 10.1242/jcs.222984.
  2. Martinez, J. (2018) Curr Opin Immunol 55, 54-61.
  3. Upadhyay, S. and Philips, J.A. (2019) Curr Opin Immunol 60, 81-90.

Pathways & Proteins

Explore pathways + proteins related to this product.

Limited Uses

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For Research Use Only. Not For Use In Diagnostic Procedures.
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