WB
Western blot analysis of Phospho-SQSTM1 / p62 (S349) on different lysates with Rabbit anti-Phospho-SQSTM1 / p62 (S349) antibody at 1/2,000 dilution. Lane 1: HeLa cell lysate, Lane 2: HeLa treated with 2μM MG-132 for 18 hours cell lysate, Lane 3: NIH/3T3 cell lysate, Lane 4: NIH/3T3 treated with 10μM MG-132 for 8 hours cell lysate, Lysates/proteins at 20 µg/Lane. Exposure time: 11 seconds; 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 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded human cerebrum tissue with Rabbit anti-Phospho-SQSTM1 / p62 (S349) antibody at 1/30,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 1/30,000 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 NIH/3T3 cells treated with 10μM MG-132 for 8 hours labeling Phospho-SQSTM1 / p62 (S349) with Rabbit anti-Phospho-SQSTM1 / p62 (S349) antibody at 1/5,000 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-Phospho-SQSTM1 / p62 (S349) antibody at 1/5,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (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 (594) was used as the secondary antibody at 1/1,000 dilution.| Product Name | Phospho-SQSTM1 / p62 (S349) Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Synthetic phospho-peptide corresponding to residues surrounding Ser349 of Human SQSTM1/ p62. |
| Modification | p-S349 |
| Clonality | monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | ICC/IFIHCWB |
| WB1:2000-1:5000IHC:1:5000 ICC/IF:1:30000 |
|
| Species Reactivity | HumanMouse |
| 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 Phospho-SQSTM1 / p62 (S349) on different lysates with Rabbit anti-Phospho-SQSTM1 / p62 (S349) antibody at 1/2,000 dilution. Lane 1: HeLa cell lysate, Lane 2: HeLa treated with 2μM MG-132 for 18 hours cell lysate, Lane 3: NIH/3T3 cell lysate, Lane 4: NIH/3T3 treated with 10μM MG-132 for 8 hours cell lysate, Lysates/proteins at 20 µg/Lane. Exposure time: 11 seconds; 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 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded human cerebrum tissue with Rabbit anti-Phospho-SQSTM1 / p62 (S349) antibody at 1/30,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 1/30,000 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 NIH/3T3 cells treated with 10μM MG-132 for 8 hours labeling Phospho-SQSTM1 / p62 (S349) with Rabbit anti-Phospho-SQSTM1 / p62 (S349) antibody at 1/5,000 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-Phospho-SQSTM1 / p62 (S349) antibody at 1/5,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (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 (594) was used as the secondary antibody at 1/1,000 dilution.| Application Notes | WB1:2000-1:5000IHC:1:5000 ICC/IF:1:30000 |
|---|
| 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. |
