SQSTM1/p62 Recombinant Antibody

$Blocking buffer: 5% NFDM\/TBST\r\nPrimary Ab dilution: 1:10000\r\nPrimary Ab incubation condition: 2 hours at \r\nroom temperature\r\nSecondary Ab: Goat Anti-Rabbit IgG H&L \r\n(HRP)\r\nLysate: (-): HCT-116, (+): HCT-116 \r\n+chloroquine (50 mM, 12h)\r\nProtein loading quantity: 20 \u03bcg\r\nExposure time: 30 s\r\nPredicted MW: 48 kDa\r\nObserved MW: 62 kDa$

Blocking buffer: 5% NFDM\/TBST\r\nPrimary Ab dilution: 1:10000\r\nPrimary Ab incubation condition: 2 hours at \r\nroom temperature\r\nSecondary Ab: Goat Anti-Rabbit IgG H&L \r\n(HRP)\r\nLysate: (-): HCT-116, (+): HCT-116 \r\n+chloroquine (50 mM, 12h)\r\nProtein loading quantity: 20 \u03bcg\r\nExposure time: 30 s\r\nPredicted MW: 48 kDa\r\nObserved MW: 62 kDa

$Blocking buffer: 5% NFDM\/TBST\r\nPrimary Ab dilution: 1:10000\r\nPrimary Ab incubation condition: 2 hours at \r\nroom temperature\r\nSecondary Ab: Goat Anti-Rabbit IgG H&L \r\n(HRP)\r\nLysate: (-): HeLa, (+): HeLa+chloroquine (50\r\nmM, 12h)\r\nProtein loading quantity: 20 \u03bcg\r\nExposure time: 30 s\r\nPredicted MW: 48 kDa\r\nObserved MW: 62 kDa$

Blocking buffer: 5% NFDM\/TBST\r\nPrimary Ab dilution: 1:10000\r\nPrimary Ab incubation condition: 2 hours at \r\nroom temperature\r\nSecondary Ab: Goat Anti-Rabbit IgG H&L \r\n(HRP)\r\nLysate: (-): HeLa, (+): HeLa+chloroquine (50\r\nmM, 12h)\r\nProtein loading quantity: 20 \u03bcg\r\nExposure time: 30 s\r\nPredicted MW: 48 kDa\r\nObserved MW: 62 kDa

$Tissue: Human colon cancer\r\nSection type: Formalin-fixed & Paraffin\u0002embedded section \r\nRetrieval method: High temperature and high \r\npressure\r\nRetrieval buffer: Tris\/EDTA buffer, pH 9.0 \r\nPrimary Ab dilution: 1:500\r\nPrimary Ab incubation condition: 1 hour at \r\nroom temperature \r\nSecondary Ab: Anti-Rabbit and Mouse \r\nPolymer HRP (Ready to use)\r\nCounter stain: Hematoxylin (Blue)\r\nComment: Color brown is the positive signal \r\nfor bsm-60709R\r\n$

Tissue: Human colon cancer\r\nSection type: Formalin-fixed & Paraffin\u0002embedded section \r\nRetrieval method: High temperature and high \r\npressure\r\nRetrieval buffer: Tris\/EDTA buffer, pH 9.0 \r\nPrimary Ab dilution: 1:500\r\nPrimary Ab incubation condition: 1 hour at \r\nroom temperature \r\nSecondary Ab: Anti-Rabbit and Mouse \r\nPolymer HRP (Ready to use)\r\nCounter stain: Hematoxylin (Blue)\r\nComment: Color brown is the positive signal \r\nfor bsm-60709R\r\n

$Tissue: Human lung squamous carcinoma\r\nSection type: Formalin-fixed & Paraffin\u0002embedded section \r\nRetrieval method: High temperature and high \r\npressure\r\nRetrieval buffer: Tris\/EDTA buffer, pH 9.0 \r\nPrimary Ab dilution: 1:500\r\nPrimary Ab incubation condition: 1 hour at \r\nroom temperature \r\nSecondary Ab: Anti-Rabbit and Mouse \r\nPolymer HRP (Ready to use)\r\nCounter stain: Hematoxylin (Blue)\r\nComment: Color brown is the positive signal \r\nfor bsm-60709R$

Tissue: Human lung squamous carcinoma\r\nSection type: Formalin-fixed & Paraffin\u0002embedded section \r\nRetrieval method: High temperature and high \r\npressure\r\nRetrieval buffer: Tris\/EDTA buffer, pH 9.0 \r\nPrimary Ab dilution: 1:500\r\nPrimary Ab incubation condition: 1 hour at \r\nroom temperature \r\nSecondary Ab: Anti-Rabbit and Mouse \r\nPolymer HRP (Ready to use)\r\nCounter stain: Hematoxylin (Blue)\r\nComment: Color brown is the positive signal \r\nfor bsm-60709R

$Paraformaldehyde-fixed, paraffin embedded (mouse spleen); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Incubation with (SQSTM1\/p62) Monoclonal Antibody, Unconjugated (bsm-60709R) at 1:200 overnight at 4\u00b0C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.$

Paraformaldehyde-fixed, paraffin embedded (mouse spleen); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Incubation with (SQSTM1\/p62) Monoclonal Antibody, Unconjugated (bsm-60709R) at 1:200 overnight at 4\u00b0C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

$Paraformaldehyde-fixed, paraffin embedded (rat spleen); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Incubation with (SQSTM1\/p62) Monoclonal Antibody, Unconjugated (bsm-60709R) at 1:200 overnight at 4\u00b0C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.$

Paraformaldehyde-fixed, paraffin embedded (rat spleen); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Incubation with (SQSTM1\/p62) Monoclonal Antibody, Unconjugated (bsm-60709R) at 1:200 overnight at 4\u00b0C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

$Paraformaldehyde-fixed, paraffin embedded (human spleen); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Incubation with (SQSTM1\/p62) Monoclonal Antibody, Unconjugated (bsm-60709R) at 1:200 overnight at 4\u00b0C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.$

Paraformaldehyde-fixed, paraffin embedded (human spleen); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Incubation with (SQSTM1\/p62) Monoclonal Antibody, Unconjugated (bsm-60709R) at 1:200 overnight at 4\u00b0C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

$Paraformaldehyde-fixed, paraffin embedded (human stomach); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Incubation with (SQSTM1\/p62) Monoclonal Antibody, Unconjugated (bsm-60709R) at 1:200 overnight at 4\u00b0C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.$

Paraformaldehyde-fixed, paraffin embedded (human stomach); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Incubation with (SQSTM1\/p62) Monoclonal Antibody, Unconjugated (bsm-60709R) at 1:200 overnight at 4\u00b0C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

$Paraformaldehyde-fixed, paraffin embedded (human liver carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Incubation with (SQSTM1\/p62) Monoclonal Antibody, Unconjugated (bsm-60709R) at 1:200 overnight at 4\u00b0C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.$

Paraformaldehyde-fixed, paraffin embedded (human liver carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Incubation with (SQSTM1\/p62) Monoclonal Antibody, Unconjugated (bsm-60709R) at 1:200 overnight at 4\u00b0C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

$25 ug total protein per lane of various lysates (see on figure) probed with SQSTM1\/p62 monoclonal antibody, unconjugated (bsm-60709R) at 1:5000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.$

25 ug total protein per lane of various lysates (see on figure) probed with SQSTM1\/p62 monoclonal antibody, unconjugated (bsm-60709R) at 1:5000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.

$Hela cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37\u00b0C for 20 min; Antibody incubation with (SQSTM1\/p62) monoclonal Antibody, Unconjugated (bsm-60709R) 1:100, 90 minutes at 37\u00b0C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37\u00b0C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.\r\n$

Hela cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37\u00b0C for 20 min; Antibody incubation with (SQSTM1\/p62) monoclonal Antibody, Unconjugated (bsm-60709R) 1:100, 90 minutes at 37\u00b0C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37\u00b0C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.\r\n

$The Hela (H) cells were fixed with 4% PFA (10 min at r.t.) and then permeabilized with 90% ice-cold methanol for 20 min at -20\u2103. Primary Antibody (green): Rabbit Anti-SQSTM1\/p62 antibody (bsm-60709R): 1 \u03bcg\/10^6 cells; Secondary Antibody (white blue): Goat anti-Rabbit IgG-FITC (bs-0295G-FITC): 1 \u03bcg\/test. Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.$

The Hela (H) cells were fixed with 4% PFA (10 min at r.t.) and then permeabilized with 90% ice-cold methanol for 20 min at -20\u2103. Primary Antibody (green): Rabbit Anti-SQSTM1\/p62 antibody (bsm-60709R): 1 \u03bcg\/10^6 cells; Secondary Antibody (white blue): Goat anti-Rabbit IgG-FITC (bs-0295G-FITC): 1 \u03bcg\/test. Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.

Applications

WB
IHC-P
IHC-F
IF

Predicted Reactivity

Human

Overview
Catalog #	bsm-60709r-100ul
Product Name	SQSTM1/p62 Recombinant Antibody
Applications	WB, IHC-P, IHC-F, IF
Predicted Reactivity	Human
Specifications
Conjugation	Unconjugated
Host	Rabbit
Source	KLH conjugated synthetic peptide derived from human SQSTM1/p62
Clonality	Recombinant
Isotype	IgG
Concentration	1ug/ul
Purification	Purified by Protein A.
Storage Buffer	0.01M TBS(pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
Storage Condition	Shipped at 4℃. Store at -20℃ for one year. Avoid repeated freeze/thaw cycles.
Target
Gene ID	8878
Swiss Prot	Q13501
Subcellular location	Cytoplasm, Nucleus
Synonyms	SQSTM_HUMAN; Sequestosome-1; ORCA; OSIL; EBI3-associated protein of 60 kDa (EBIAP; p60); Phosphotyrosine-independent ligand for the Lck SH2 domain of 62 kDa; Ubiquitin-binding protein p62;
Application Dilution
WB	1:300-5000
IHC-P	1:200-400
IHC-F	1:100-500
IF