Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® SQSTM1/p62 siRNA I #6394 (+) or SignalSilence® SQSTM1/p62 siRNA II #6399 (+), using SQSTM1/p62 (D5E2) Rabbit mAb (upper) or α-Tubulin (11H10) Rabbit mAb #2125 (lower). The SQSTM1/p62 (D5E2) Rabbit mAb confirms silencing of SQSTM1/p62 expression, while the α-Tubulin (11H10) Rabbit mAb is used as a loading control.
Western blot analysis of extracts from HeLa cells (lane 1) or SQSTM1 knock-out cells (lane 2) using SQSTM1/p62 (D5E2) Rabbit mAb #8025 (upper), and β-actin (D6A8) Rabbit mAb #8457 (lower). The absence of signal in the SQSTM1 knock-out HeLa cells confirms specificity of the antibody for SQSTM1.
Western blot analysis of extracts from various cell lines using Optineurin (D2L8S) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of extracts from various cell lines using NDP52 (D1E4A) Rabbit mAb.
Western blot analysis of extracts from various cell lines using TAX1BP1 (D1D5) Rabbit mAb.
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.
Western blot analysis of extracts from 293T cells, either mock transfected (-) or transfected with a Myc/DDK-tagged cDNA expression construct encoding the full-length isoform 1 of human TAX1BP1 (hTAX1BP1-Myc/DDK, +), using TAX1BP1 (D1D5) Rabbit mAb.
Western blot analysis of extracts from various cell lines using SQSTM1/p62 (D5E2) Rabbit mAb.
Western blot analysis of extracts from SK-MEL-2 cells, untreated (-) or starved overnight in Earle's Balanced Salt Solution (EBSS) (+), using SQSTM1/p62 (D5E2) Rabbit mAb.
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a tagged human SQSTM1/p62 construct (+), using SQSTM1/p62 (D5E2) Rabbit mAb.
The SQSTM1/p62-like Receptor Antibody Sampler Kit provides an economical means of detecting members of the SQSTM1/p62-like Receptor (SLR) family. The kit includes enough antibody to perform two western blot experiments with each primary antibody.
Each antibody in the SQSTM1/p62-like Receptor Antibody Sampler Kit detects endogenous levels of its target protein. Based on sequence alignment, TAX1BP (D1D5) Rabbit mAb is predicted to react with TXBP151-L and TXBP151-S isoforms.
Monoclonal antibodies are produced by immunizing rabbits with synthetic peptides corresponding to Gly162 of human SQSTM1/p62, Lys601 of human NBR1, Leu410 of human Optineurin, Val135 of NDP52, and residues near the carboxy terminus of human TAX1BP1.
Autophagy is a catabolic process for the autophagosome-lysosomal degradation of bulk cytoplasmic contents (1,2). Selective autophagy targets the degradation of distinct sets of substrates and organelles and can occur through the utilization of a number of autophagy cargo receptors (3-5). Autophagy cargo receptors contain an LC3-interacting region (LIR) required for interaction with Atg8/LC3 family members targeted to the autophagosome. SQSTM1/p62-like receptors (SLRs) are a family of autophagy cargo receptors that contain domains for binding to ubiquitin. This family includes prototypical member SQSTM1/p62, NBR1, NDP52, Optineurin, and TAX1BP1. Targets of SLRs include ubiquitylated protein aggregates (aggrephagy), organelles such as mitochondria (mitoophagy) and peroxisomes (pexophagy), and intracellular bacteria (xenophagy).
Upon binding of cargo to these receptors, the complex is delivered to the autophagosome where both the cargo and receptor are degraded through the autophagic process. While some redundancy may exist among SLR family members, they can have unique activities. Many SLRs can have additional roles as scaffolding proteins for various signaling pathways. For example, SQSTM1/p62 interacts with KEAP1, a cytoplasmic inhibitor of NRF2, a key transcription factor involved in cellular responses to oxidative stress (6). Thus, accumulation of SQSTM1/p62 can lead to an increase in NRF2 activity.
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