SQSTM1 / p62 Recombinant Rabbit Monoclonal Antibody [PS00-61]

RMB: 900 特惠 1500 1500

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100μl

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Catalog# HA721171

SQSTM1 / p62 Recombinant Rabbit Monoclonal Antibody [PS00-61]

Application

WB
IHC-P
IF-Cell
FC
IF-Tissue
IHC-Fr

Reactivity

Human
Mouse
Rat

Predicted reactivity

Cynomolgus monkey
Pig

This product has been cited in peer reviewed publications, see list HERE

概述

产品名称

SQSTM1 / p62 Recombinant Rabbit Monoclonal Antibody [PS00-61]

抗体类型

Recombinant Rabbit monoclonal Antibody

免疫原

Synthetic peptide within Human SQSTM1/ p62 aa 400 to the C-terminus

种属反应性

Human, Mouse, Rat (Predicted: Cynomolgus monkey, Pig)

验证应用

WB, IHC-P, IF-Cell, FC, IF-Tissue, IHC-Fr

分子量

Predicted band size: 48 kDa

阳性对照

HeLa cell lysate, HeLa treated with 50μM Chloroquine for 18 hours cell lysate, NIH/3T3 cell lysate, C2C12 cell lysate, PC-12 cell lysate, PANC-1 cell lysate, RAW264.7 cell lysate, Mouse liver tissue lysate, C2C12, PC-12, A549, human colon carcinoma tissue, human liver tissue, mouse liver tissue, rat liver tissue, mouse brain tissue, rat brain tissue.

偶联

unconjugated

克隆号

PS00-61

RRID

AB_3072293

产品特性

形态

Liquid

存放说明

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

存储缓冲液

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

亚型

IgG

纯化方式

Protein A affinity purified.

应用稀释度

WB
1:5,000-1:50,000

IHC-P
1:1,000-1:2,000

IF-Cell
1:100-1:1,000

FC
1:1,000

IF-Tissue
1:500-1:1,000

IHC-Fr
1:500

发表文章中的应用

WB	查看 53 篇文献如下
IF	查看 3 篇文献如下
IHC-P	查看 1 篇文献如下
IF-cell	查看 1 篇文献如下
mIHC	查看 1 篇文献如下
IF-tissue	查看 1 篇文献如下

发表文章中的种属

Mouse	查看 29 篇文献如下
Human	查看 20 篇文献如下
Rat	查看 6 篇文献如下
Grass carp	查看 1 篇文献如下
Pig	查看 1 篇文献如下
Chicken	查看 1 篇文献如下
largemouth bass	查看 1 篇文献如下
Ctenopharyngodon	查看 1 篇文献如下

靶点

功能

Autophagy receptor required for selective macroautophagy (aggrephagy). Functions as a bridge between polyubiquitinated cargo and autophagosomes. Interacts directly with both the cargo to become degraded and an autophagy modifier of the MAP1 LC3 family. Along with WDFY3, involved in the formation and autophagic degradation of cytoplasmic ubiquitin-containing inclusions (p62 bodies, ALIS/aggresome-like induced structures). Along with WDFY3, required to recruit ubiquitinated proteins to PML bodies in the nucleus. May regulate the activation of NFKB1 by TNF-alpha, nerve growth factor (NGF) and interleukin-1. May play a role in titin/TTN downstream signaling in muscle cells. May regulate signaling cascades through ubiquitination. Adapter that mediates the interaction between TRAF6 and CYLD. May be involved in cell differentiation, apoptosis, immune response and regulation of K+ channels. Involved in endosome organization by retaining vesicles in the perinuclear cloud: following ubiquitination by RNF26, attracts specific vesicle-associated adapters, forming a molecular bridge that restrains cognate vesicles in the perinuclear region and organizes the endosomal pathway for efficient cargo transport. Promotes relocalization of 'Lys-63'-linked ubiquitinated STING1 to autophagosomes. Acts as an activator of the NFE2L2/NRF2 pathway via interaction with KEAP1: interaction inactivates the BCR(KEAP1) complex, promoting nuclear accumulation of NFE2L2/NRF2 and subsequent expression of cytoprotective genes.

背景文献

1. Tan C.T., Chang H.C., Zhou Q., Yu C., Fu N.Y., Sabapathy K., Yu V.C. MOAP-1-mediated dissociation of p62/SQSTM1 bodies releases Keap1 and suppresses Nrf2 signaling. EMBO Rep. 22:e50854-e50854(2021)

2. Prabakaran T., Bodda C., Krapp C., Zhang B.C., Christensen M.H., Sun C., Reinert L., Cai Y., Jensen S.B., Paludan S.R. Attenuation of cGAS-STING signaling is mediated by a p62/SQSTM1-dependent autophagy pathway activated by TBK1. EMBO J. 37:0-0(2018)

亚细胞定位

Cytoplasm, Endoplasmic reticulum, Endosome, Lysosome, Nucleus.

UNIPROT

SwissProt: Q13501 Human

SwissProt: Q64337 Mouse

SwissProt: O08623 Rat

别名

A170 antibody

DMRV antibody

EBI 3 associated protein of 60 kDa antibody

EBI 3 associated protein p60 antibody

EBI3 associated protein of 60 kDa antibody

EBI3 associated protein p60 antibody

EBI3-associated protein of 60 kDa antibody

EBIAP antibody

FTDALS3 antibody

MGC127197 antibody

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图片

☑ Cell treatment (CT)

Western blot analysis of SQSTM1 / p62 on different lysates with Rabbit anti-SQSTM1 / p62 antibody (HA721171) at 1/20,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: HeLa treated with 50μM Chloroquine for 18 hours cell lysate
Lane 3: NIH/3T3 cell lysate
Lane 4: C2C12 cell lysate
Lane 5: PC-12 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 48 kDa
Observed band size: 62 kDa

Exposure time: 2 seconds; ECL: K1801;
4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721171) at 1/20,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
Western blot analysis of SQSTM1 / p62 on different lysates with Rabbit anti-SQSTM1 / p62 antibody (HA721171) at 1/5,000 dilution.

Lane 1: PANC-1 cell lysate (10 µg/Lane)
Lane 2: RAW264.7 cell lysate (10 µg/Lane)
Lane 3: Mouse liver tissue lysate (20 µg/Lane)

Predicted band size: 62 kDa
Observed band size: 62 kDa

Exposure time: 50 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721171) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
☑ Knockdown (KD)

Western blot analysis of SQSTM1 / p62 on different lysates with Rabbit anti-SQSTM1 / p62 antibody (HA721171) at 1/20,000 dilution.

Lane 1: A549-si NT cell lysate
Lane 2: A549-si SQSTM1 / p62 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 48 kDa
Observed band size: 62 kDa

Exposure time: 17 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721171) at 1/20,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
Immunocytochemistry analysis of C2C12 cells labeling SQSTM1 / p62 with Rabbit anti-SQSTM1 / p62 antibody (HA721171) at 1/1,000 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-SQSTM1 / p62 antibody (HA721171) at 1/1,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
Immunocytochemistry analysis of PC-12 cells labeling SQSTM1 / p62 with Rabbit anti-SQSTM1 / p62 antibody (HA721171) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-SQSTM1 / p62 antibody (HA721171) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
Immunocytochemistry analysis of A549 cells labeling SQSTM1 / p62 with Rabbit anti-SQSTM1 / p62 antibody (HA721171) at 1/50 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-SQSTM1 / p62 antibody (HA721171) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Application: IHC-Fr

Species: Mouse

Site: Cerebral cortex

Sample: Frozen section

Antibody concentration: 1/500

Antigen retrieval: The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven.
Application: IF-tissue

Species: Mouse

Site: Liver

Sample: Paraffin-embedded section

Antibody concentration: 1/500
Application: IF-tissue

Species: Mouse

Site: brain

Sample: Paraffin-embedded section

Antibody concentration: 1/500
Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-SQSTM1 / p62 antibody (HA721171) at 1/400 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA721171) at 1/400 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-SQSTM1 / p62 antibody (HA721171) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA721171) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-SQSTM1 / p62 antibody (HA721171) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA721171) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded rat liver tissue with Rabbit anti-SQSTM1 / p62 antibody (HA721171) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA721171) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-SQSTM1 / p62 antibody (HA721171) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA721171) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-SQSTM1 / p62 antibody (HA721171) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA721171) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-SQSTM1 / p62 antibody (HA721171) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA721171) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Flow cytometric analysis of PC-12 cells labeling SQSTM1 / p62.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA721171, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).