WB
Western blot analysis of SQSTM1 / p62 on different lysates with Rabbit anti-SQSTM1 / p62 antibody at 1/5,000 dilution. Lane 1: SH-SY5Y cell lysate, Lane 2: PANC-1 cell lysate, Lane 3: HeLa cell lysate, Lysates/proteins at 10 µg/Lane. Exposure time: 1 minute; 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 at 1/5,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:100,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-SQSTM1 / p62 antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 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.ICC/IF
Immunocytochemistry analysis of C2C12 cells labeling SQSTM1 / p62 with Rabbit anti-SQSTM1 / p62 antibody 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 at 1/1,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.FC
Flow cytometric analysis of PC-12 cells labeling SQSTM1 / p62. Cells were fixed and permeabilized. Then stained with the primary antibody (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 at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Product Name | SQSTM1 / p62 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Synthetic peptide within Human SQSTM1/ p62 aa 400 to the C-terminus. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFIHCWB |
| WB:1:5000-1:10000 IHC:1:1000-1:2000 ICC:1:100-1:1000 FC:1:1000 |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | SQSTM1 |
|---|---|
| Gene Synonyms | p60 p62 A170 DMRV OSIL PDB3 ZIP3 p62B EBIAP NADGP FTDALS3 |
| Gene Full Name | sequestosome 1 |
| Gene Summary | This gene encodes a multifunctional protein that binds ubiquitin and regulates activation of the nuclear factor kappa-B (NF-kB) signaling pathway. The protein functions as a scaffolding/adaptor protein in concert with TNF receptor-associated factor 6 to mediate activation of NF-kB in response to upstream signals. Alternatively spliced transcript variants encoding either the same or different isoforms have been identified for this gene. Mutations in this gene result in sporadic and familial Paget disease of bone. [provided by RefSeq, Mar 2009] |
| Alternative Names | 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 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 ORCA antibody OSF-6 antibody Osi antibody OSIL antibody Oxidative stress induced like antibody p60 antibody p62 antibody p62B antibody Paget disease of bone 3 antibody PDB 3 antibody PDB3 antibody Phosphotyrosine independent ligand for the Lck SH2 domain of 62 kDa antibody Phosphotyrosine independent ligand for the Lck SH2 domain p62 antibody Phosphotyrosine-independent ligand for the Lck SH2 domain of 62 kDa antibody PKC-zeta-interacting protein antibody Protein kinase C-zeta-interacting protein antibody Sequestosome 1 antibody Sequestosome-1 antibody SQSTM 1 antibody SQSTM_HUMAN antibody Sqstm1 antibody STAP antibody STONE14 antibody Ubiquitin binding protein p62 antibody Ubiquitin-binding protein p62 antibody ZIP 3 antibody ZIP antibody ZIP3 antibody |
| Molecular Weight(MW) | 48kDa(Observed band size: 62kDa) |
| Cellular Localization | Cytoplasm, Endoplasmic reticulum, Endosome, Lysosome, Nucleus. |
WB
Western blot analysis of SQSTM1 / p62 on different lysates with Rabbit anti-SQSTM1 / p62 antibody at 1/5,000 dilution. Lane 1: SH-SY5Y cell lysate, Lane 2: PANC-1 cell lysate, Lane 3: HeLa cell lysate, Lysates/proteins at 10 µg/Lane. Exposure time: 1 minute; 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 at 1/5,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:100,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-SQSTM1 / p62 antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 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.
ICC/IF
Immunocytochemistry analysis of C2C12 cells labeling SQSTM1 / p62 with Rabbit anti-SQSTM1 / p62 antibody 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 at 1/1,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.
FC
Flow cytometric analysis of PC-12 cells labeling SQSTM1 / p62. Cells were fixed and permeabilized. Then stained with the primary antibody (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 at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Application Notes | WB:1:5000-1:10000 IHC:1:1000-1:2000 ICC:1:100-1:1000 FC:1:1000 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
