SQSTM1/p62 Recombinant Mouse Monoclonal Antibody

货号：

AWA00001

应用：

WB,IHC-P,IF-C,IF-T,FCM

反应性：

Human,Mouse,Rat

来源：

Mouse

规格 价格 库存
20μL ¥620 1-3个工作日
50μL ¥1250 1-3个工作日
100μL ¥2200 1-3个工作日

产品概述

Product Details

Host Species:

Mouse

Reactivity:

Human, Mouse, Rat

Molecular Wt:

Predicted MW: 48 kDa
Observed MW: 62 kDa

Clonality:

Monoclonal

Isotype:

IgG2a

Concentration:

0.568mg/ml

Other Names:

P62; p60; p62B; EBIAP; FTDALS3; STAP; STONE14; ORCA; OSF-6; Osi; OSIL; ZIP 3; ZIP3; ZIP; Sequestosome 1; Sequestosome-1; PDB 3; PDB3; SQSTM 1; SQSTM1; Ubiquitin binding protein p62; SQSTM1/p62

Formulation:

Liquid in PBS containing 50% glycerol, 0.5% BSA and 0.02% sodium azide.

Purification:

Affinity-chromatography

Storage:

Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20°C storage.

Applications

WB 1:1000-1:5000
IHC-P 1:100-1:1000
IF-C 1:100-1:800
IF-T 1:100-1:1000
FCM 1:50-1:200

Immunogen
Information

Gene Name:

SQSTM1

Protein Name:

Sequestosome-1

Gene ID:

8878 (Human)
18412 (Mouse)
113894 (Rat)

SwissPro:

Q13501 (Human)
Q64337 (Mouse)
O08623 (Rat)

Immunogen
Information

Subcellular Location:

Cytoplasmic vesicle, autophagosome. Preautophagosomal structure. Cytoplasm, cytosol. Nucleus, PML body. Late endosome. Lysosome. Nucleus. Endoplasmic reticulum. Cytoplasm, myofibril, sarcomere.

Immunogen:

Synthetic peptide within human SQSTM1/p62. AA range: 400 to the C-terminus.

Specificity:

SQSTM1/p62 Monoclonal Antibody detects endogenous levels of SQSTM1/p62 protein.

Product images
	Fig : Western blot analysis of SQSTM1/p62 on different lysates. Proteins were transferred to a NC membrane and blocked with 5% NF-Milk in TBST for 1 hour at room temperature. The primary antibody (AWA00001, 1/1000) was used in TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (AWS0001) at 1:5,000 dilution was used for 1 hour at room temperature. Positive control: Lane 1: PANC-1 cell Lane 2: C2C12 cell Lane 3: SK-BR-3 cell Lane 4: CTX cell Lane 5: PC12 cell Predicted molecular weight:48 kDa Observed molecular weight:62 kDa

	Fig : Immunohistochemical analysis of paraffin-embedded Mouse-colon tissue with mouse anti-SQSTM1/p62 antibody (AWA00001) at 1/100 dilution. The section was pre-treated using heat mediated antigen retrieval with Sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 3% H₂O₂ for 15 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (AWA00001) at 1/100 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system(ABIOWELL, AWI0629). DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

	Fig: Fluorescence immunohistochemical analysis of Mouse-colon tissue (Formalin / PFA-fixed paraffin-embedded sections) with mouse anti-SQSTM1/p62 antibody (AWA00001) at 1/100 dilution. The immunostaining was performed with the TSA Immuno-staining Kit (ABIOWELL, AWI0689). The section was pre-treated using heat mediated antigen retrieval with EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 5% BSA for 60 minutes at 37℃, washed with ddH2O and PBS, and then probed with the primary antibody (AWA00001) at 1/100 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system followed by a separate fluorescent tyramide signal amplification system (red). DAPI (blue, AWC0291) was used as a nuclear counter stain. Image acquisition was performed with Slide Scanner.

	Fig : Western blot analysis of SQSTM1 on different lysates. Proteins were transferred to a NC membrane and blocked with 5% NF-Milk in TBST for 90 minutes at room temperature. The primary antibody (AWA00001, 1/1000) was used in TBST at room temperature for 90 minutes. Goat Anti-Mouse IgG - HRP Secondary Antibody (AWS0001) at 1:5,000 dilution was used for 1 hour at room temperature. Positive control: Lane 1: HEK293 cell Predicted molecular weight:48 kDa Observed molecular weight:62 kDa

	Fig: Fluorescence immunohistochemical analysis of Mouse-cortex tissue (Formalin / PFA-fixed paraffin-embedded sections) with mouse anti-SQSTM1/p62 antibody (AWA00001) at 1/100 dilution. The immunostaining was performed with the TSA Immuno-staining Kit (ABIOWELL, AWI0689). The section was pre-treated using heat mediated antigen retrieval with EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 5% BSA for 60 minutes at 37℃, washed with ddH2O and PBS, and then probed with the primary antibody (AWA00001) at 1/100 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system followed by a separate fluorescent tyramide signal amplification system (red). DAPI (blue, AWC0291) was used as a nuclear counter stain. Image acquisition was performed with Slide Scanner.

	Fig: Fluorescence immunohistochemical analysis of Rat-colon tissue (Formalin / PFA-fixed paraffin-embedded sections) with mouse anti-SQSTM1/p62 antibody (AWA00001) at 1/100 dilution. The immunostaining was performed with the TSA Immuno-staining Kit (ABIOWELL, AWI0689). The section was pre-treated using heat mediated antigen retrieval with EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 5% BSA for 60 minutes at 37℃, washed with ddH2O and PBS, and then probed with the primary antibody (AWA00001) at 1/100 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system followed by a separate fluorescent tyramide signal amplification system (red). DAPI (blue, AWC0291) was used as a nuclear counter stain. Image acquisition was performed with Slide Scanner.

	Fig: Fluorescence immunohistochemical analysis of Mouse-hippocampus tissue (Formalin / PFA-fixed paraffin-embedded sections) with mouse anti-SQSTM1/p62 antibody (AWA00001) at 1/100 dilution. The immunostaining was performed with the TSA Immuno-staining Kit (ABIOWELL, AWI0689). The section was pre-treated using heat mediated antigen retrieval with EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 5% BSA for 60 minutes at 37℃, washed with ddH2O and PBS, and then probed with the primary antibody (AWA00001) at 1/100 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system followed by a separate fluorescent tyramide signal amplification system (red). DAPI (blue, AWC0291) was used as a nuclear counter stain. Image acquisition was performed with Slide Scanner.

	Fig : Immunohistochemical analysis of paraffin-embedded Mouse-brain tissue with mouse anti-SQSTM1/p62 antibody (AWA00001) at 1/100 dilution. The section was pre-treated using heat mediated antigen retrieval with Sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 3% H₂O₂ for 15 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (AWA00001) at 1/100 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system(ABIOWELL, AWI0629). DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

	Fig : Immunohistochemical analysis of paraffin-embedded Mouse-kidney tissue with mouse anti-SQSTM1/p62 antibody (AWA00001) at 1/100 dilution. The section was pre-treated using heat mediated antigen retrieval with Sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 3% H₂O₂ for 15 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (AWA00001) at 1/100 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system(ABIOWELL, AWI0629). DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

	Fig : Immunohistochemical analysis of paraffin-embedded Rat-brain tissue with mouse anti-SQSTM1/p62 antibody (AWA00001) at 1/100 dilution. The section was pre-treated using heat mediated antigen retrieval with Sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 3% H₂O₂ for 15 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (AWA00001) at 1/100 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system(ABIOWELL, AWI0629). DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

	Fig: Fluorescence immunohistochemical analysis of Rat-cortex tissue (Formalin / PFA-fixed paraffin-embedded sections) with mouse anti-SQSTM1/p62 antibody (AWA00001) at 1/100 dilution. The immunostaining was performed with the TSA Immuno-staining Kit (ABIOWELL, AWI0689). The section was pre-treated using heat mediated antigen retrieval with EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 5% BSA for 60 minutes at 37℃, washed with ddH2O and PBS, and then probed with the primary antibody (AWA00001) at 1/100 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system followed by a separate fluorescent tyramide signal amplification system (red). DAPI (blue, AWC0291) was used as a nuclear counter stain. Image acquisition was performed with Slide Scanner.

	Fig : Immunohistochemical analysis of paraffin-embedded Rat-kidney tissue with mouse anti-SQSTM1/p62 antibody (AWA00001) at 1/100 dilution. The section was pre-treated using heat mediated antigen retrieval with Sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 3% H₂O₂ for 15 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (AWA00001) at 1/100 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system(ABIOWELL, AWI0629). DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

	Fig : Immunohistochemical analysis of paraffin-embedded Rat-colon tissue with Mouse anti-SQSTM1/p62 antibody (AWA00001) at 1/100 dilution. The section was pre-treated using heat mediated antigen retrieval with Sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 3% H₂O₂ for 15 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (AWA00001) at 1/100 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system(ABIOWELL, AWI0629). DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

引用文献 (1)

FUNCTIONAL & INTEGRATIVE GENOMICS IF:3.9

SIGMAR1 targets AMPK/ULK1 pathway to inhibit SH-SY5Y cell apoptosis by regulating endoplasmic reticulum stress and autophagy

Distal hereditary motor neuropathy (dHMN) is a progressive neurological disease characterized by distal limb muscle weakness and amyotrophy. Sigma 1 receptor (σ1R), a gene product of SIGMAR1, mutations have been reported to induce dHMN, but its mechanism remains unknown. This study aims to explore the effect of C238T and 31_50del mutations in σ1R on neuronal SH-SY5Y cell functions. The SH-SY5Y cells that overexpressed σ1R, C238T mutant σ1R (σ1R C238T ) or 31_50del mutant σ1R (σ1R 31_50del ) were constructed by pEGFPN1 vectors. We used Western blot (WB) and immunofluorescence (IF) staining to detect the expression of σ1R and green fluorescent proteins (GFP). Then, we evaluated the impact of σ1R mutation on apoptosis, autophagy, endoplasmic reticulum stress, and the involvement of the unfolded protein response (UPR) pathway in SH-SY5Y cells. We found that σ1R C238T and σ1R 31_50del downregulated σ1R and promoted the apoptosis of SH-SY5Y cells. σ1R C238T and σ1R 31_50del increased p-PERK, p-eIF2α, p-JNK, BIP, ATF4, CHOP, ATF6, XBP1, Caspase3, Caspase12 expressions and Ca 2+ concentration, whereas decreased ATP content in SH-SY5Y cells. Besides, the expressions of LC3B, Lamp1, ATG7, Beclin-1 and phosphorylation of AMPK and ULK1 were increased, while the p62 level decreased after C238T or 31_50del mutation of σ1R. Additionally, AMPK knockdown abolished the apoptosis mediated by σ1R C238T or σ1R 31_50del in SH-SY5Y cells. Our results indicated that C238T or 31_50del mutation in σ1R promoted motor neuron apoptosis through the AMPK/ULK1 pathway in dHMN. This study shed light on a better understanding of the neurons pathological mechanisms mediated by σ1R C238T and σ1R 31-50del in dHMN.

pubTime 2024-08-07

Application

Specie

Human

Dilution

1:4000