抗p62/SQSTM1抗体 | Anti- p62/SQSTM1 antibody

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	Biological Strategies Validation. Western Blot: p62/SQSTM1 Antibody [NBP1-48320] - Cultured HeLa cells were treated with or without 50 uM chloriquine for 24 hours as indicated. Cell lysates were prepared and separated on a 12% gel by SDS-PAGE. Protein was transferred to PVDF mebbrane and blocked in 5% non-fat milk. The membrane was then probed with 1 ug/ml anti-p62/SQSMT1 in 1% milk and dectected with an anti-rabbit HRP secondary antibody using chemiluminescence. Note the upregulation of p62 (arrowhead) in response to chloroquine treatment and the blockage of autophagy.
	Biological Strategies Validation. Immunocytochemistry/Immunofluorescence: p62/SQSTM1 Antibody [NBP1-48320] - HeLa cells were treated overnight with 50uM CQ, then fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton-X100. The cells were incubated with anti-p628/SQSTM1 at a 1:200 dilution overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.
	Biological Strategies Validation. Immunocytochemistry/Immunofluorescence: p62/SQSTM1 Antibody [NBP1-48320] - Untreated HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton-X100. The cells were incubated with anti-p628/SQSTM1 at a 1:200 dilution overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.
	Immunocytochemistry/Immunofluorescence: p62/SQSTM1 Antibody [NBP1-48320] - ICC/IF analysis of formalin fixed HeLa cells using SQSTM1/p62 antibody. Blocking was performed using 10% normal goat serum and the primary antibody was used at 1:200 dilution with 4C ON incubation. The signal was developed using Dylight 488 conjugated goat-anti Rabbit secondary antibody. Cytoplasmic and punctate (autophagosomes) staining was observed which is the expected signal for p62 protein.
	Simple Western: p62/SQSTM1 Antibody [NBP1-48320] - Simple Western lane view shows a specific band for p62/SQSTM1 in 1.0 mg/ml of HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

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Background

p62/SQSTM1 (sequestosome 1) is ubiquitously-expressed cytoplasmic/adapter protein conserved in vertebrates and it can be induced by proteasomal inhibitor PSI, PGJ2/prostaglandin J2 as well as phorbol 12-myristate 13-acetate (PMA). SQSTM1 has the ability of ubiquitin binding as well as regulating NFkB1 activation by TNF-alpha, NGF (nerve growth factor) and interleukin-1. SQSTM1 acts as an adapter that mediates the interaction between TRAF6 and CYLD, and SQSTM1-TRAF6 interaction leads to K63-linked polyubiquitination of TRAF6 followed by subsequent activation of NFkB pathway. SQSTM1 plays a role in TITIN/TTN downstream signaling in muscle cells and regulate signaling cascades through ubiquitination. SQSTM1 is essential both for formation and autophagic degradation of polyubiquitin-containing bodies which are known as aggresome-like induced structures (ALIS) and SQSTM1 connects ALIS to the autophagic machinery via direct interaction with MAP1 LC3 family members. Moreover, while autophagy modulates SQSTM1 protein levels, SQSTM1 suppresses autophagy via activation of mTORC1. SQSTM1 interacts with KEAP1, which is a cytoplasmic inhibitor of NRF2, a key transcription factor involved in cellular responses to oxidative stress. SQSTM1 functions as a signaling hub for various signal transduction pathways, apoptosis, cell differentiation, apoptosis, immune response, regulation of K+ channels and Nrf2 activation, and its dysregulation is associated with Paget disease of bone and tumorigenesis.

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