Phospho-SQSTM1 / p62 (S349) Recombinant Rabbit Monoclonal Antibody [PSH08-97]
cat.: HA723050
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse
Applications: WB, IF-Cell, IHC-P
Clonality: Monoclonal
Clone number: PSH08-97
Form: Liquid
Storage condition: 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.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 48 kDa
Isotype: IgG
Immunogen: Synthetic phospho-peptide corresponding to residues surrounding Ser349 of Human SQSTM1/ p62.
Positive control: HeLa treated with 2μM MG-132 for 18 hours cell lysate, NIH/3T3 treated with 10μM MG-132 for 8 hours cell lysate, HeLa cells treated with 2μM MG-132 for 18 hours, NIH/3T3cells treated with 10μM MG-132 for 8 hours , human cerebrum tissue.
Subcellular location: Cytoplasm, Endoplasmic reticulum, Endosome, Lysosome, Nucleus.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
1:2,000-1:5,000
1:5,000
1:30,000
Uniprot #: SwissProt: Q13501 Human | Q64337 Mouse
Alternative names: A170 DMRV EBI 3 associated protein of 60 kDa EBI 3 associated protein p60 EBI3 associated protein of 60 kDa EBI3 associated protein p60 EBI3-associated protein of 60 kDa EBIAP FTDALS3 MGC127197 ORCA OSF-6 Osi OSIL Oxidative stress induced like p60 p62 p62B Paget disease of bone 3 PDB 3 PDB3 Phosphotyrosine independent ligand for the Lck SH2 domain of 62 kDa Phosphotyrosine independent ligand for the Lck SH2 domain p62 Phosphotyrosine-independent ligand for the Lck SH2 domain of 62 kDa PKC-zeta-interacting protein Protein kinase C-zeta-interacting protein Sequestosome 1 Sequestosome-1 SQSTM 1 SQSTM_HUMAN Sqstm1 STAP STONE14 Ubiquitin binding protein p62 Ubiquitin-binding protein p62 ZIP 3 ZIP ZIP3
Images
HA723050_1.jpg Fig1: Western blot analysis of Phospho-SQSTM1 / p62 (S349) on different lysates with Rabbit anti-Phospho-SQSTM1 / p62 (S349) antibody (HA723050) 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.

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

Exposure time: 11 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 (HA723050) at 1/2,000 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.
HA723050_2.jpg Fig2: Immunocytochemistry analysis of HeLa cells treated with 2μM MG-132 for 18 hours labeling Phospho-SQSTM1 / p62 (S349) with Rabbit anti-Phospho-SQSTM1 / p62 (S349) antibody (HA723050) 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 (HA723050) at 1/5,000 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 (HA601187, 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.
HA723050_3.jpg Fig3: 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 (HA723050) 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 (HA723050) at 1/5,000 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 (HA601187, 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.
HA723050_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human cerebrum tissue with Rabbit anti-Phospho-SQSTM1 / p62 (S349) antibody (HA723050) 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 (HA723050) 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.
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