WB
Western blot analysis using SQSTM1 antibody against Hela(1),Hepg2(2),Cos-7(3),A431(4),K562(5),Mcf-7(6),A549(7) cell lysate. 12% SDS-PAGE gel.Sample loading: 20μg /lane. Transfer the proteins onto a PVDF membrane (OM790003), and block it with TBST (OM750016) plus skimmed milk powder(OM750084) for one hour. Dilute the primary antibody with the antibody diluent (OM750012) at a ratio of 1:1000, and incubate it overnight at 4°C. Wash the membrane three times with TBST (OM750016), 5 minutes each time. At room temperature, dilute the secondary antibody, Goat Anti-Rabbit IgG(H&L)-HRP (OM643487), at a ratio of 1:20000 and incubate for one hour. Wash the membrane three times with TBST (OM750016) again, 5 minutes each time. Use ECL (OM625701) for luminescence.staining time: 60S.ICC/IF
Immunofluorescence analysis of HUVEC cells using SQSTM1 antibody (green). Blue: DAPI fluorescent DNA dye. Red: Actin filaments have been labeled with Omnimabs® 594-Phalloidin.Cells are fixed in 4% paraformaldehyde at room temperature for 15 minutes. Then, they are permeabilized with a PBS (OM750003) solution containing 0.1% Triton X-100(OM750021) at room temperature for 15 minutes. Subsequently, the cells are blocked with 10% non - immune goat serum(OM760028) at room temperature for 1 hour.The cells are incubated overnight at 4°C with the primary antibody diluted 1:100 in PBS. The secondary antibody, Omnimabs® 488 Goat Ant-Rabbit IgG(H&L) (Green,OM643486), is diluted at a ratio of 1:400 and incubated with the cells for 1 hour.Nuclear DNA is labeled with DAPI (Blue,OM643160). F-actin is stained with Omnimabs® 594-Phalloidin (Red,OM750007) diluted 1:100 for 30 minutes.ELISA
Coat the plate with immunizing antigen.Blue line： Immune serum; Orange line: Pre-immune serum.| Product Name | Anti-SQSTM1 antibody,pbs only |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Polypeptide |
| Clonality | Polyclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | ELISAICC/IFWB |
| WB:1:1000-1:2000 ICC/IF:1:100-1:500 |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | SQSTM1 |
|---|---|
| Gene Synonyms | A170 DMRV EBIAP FTDALS3 NADGP OSF-6 Osi OSIL p60 p62 p62B PDB3 STAP STONE14 ZIP ZIP3 |
| 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 | autophagy receptor p62 EBI3-associated protein of 60 kDa EBI3-associated protein p60 oxidative stress induced oxidative stress induced like p62 transcript variant 4 p62 transcript variant 5 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 ubiquitin-binding protein p62 |
| Molecular Weight(MW) | 48kDa(observed band size：60kDa） |
| Cellular Localization | Cytoplasm, Cytoplasmic vesicle, Endoplasmic reticulum, Endosome, Lysosome, Nucleus |
| SwissProt ID | Q13501 |
|---|
WB
Western blot analysis using SQSTM1 antibody against Hela(1),Hepg2(2),Cos-7(3),A431(4),K562(5),Mcf-7(6),A549(7) cell lysate. 12% SDS-PAGE gel.Sample loading: 20μg /lane. Transfer the proteins onto a PVDF membrane (OM790003), and block it with TBST (OM750016) plus skimmed milk powder(OM750084) for one hour. Dilute the primary antibody with the antibody diluent (OM750012) at a ratio of 1:1000, and incubate it overnight at 4°C. Wash the membrane three times with TBST (OM750016), 5 minutes each time. At room temperature, dilute the secondary antibody, Goat Anti-Rabbit IgG(H&L)-HRP (OM643487), at a ratio of 1:20000 and incubate for one hour. Wash the membrane three times with TBST (OM750016) again, 5 minutes each time. Use ECL (OM625701) for luminescence.staining time: 60S.
ICC/IF
Immunofluorescence analysis of HUVEC cells using SQSTM1 antibody (green). Blue: DAPI fluorescent DNA dye. Red: Actin filaments have been labeled with Omnimabs® 594-Phalloidin.Cells are fixed in 4% paraformaldehyde at room temperature for 15 minutes. Then, they are permeabilized with a PBS (OM750003) solution containing 0.1% Triton X-100(OM750021) at room temperature for 15 minutes. Subsequently, the cells are blocked with 10% non - immune goat serum(OM760028) at room temperature for 1 hour.The cells are incubated overnight at 4°C with the primary antibody diluted 1:100 in PBS. The secondary antibody, Omnimabs® 488 Goat Ant-Rabbit IgG(H&L) (Green,OM643486), is diluted at a ratio of 1:400 and incubated with the cells for 1 hour.Nuclear DNA is labeled with DAPI (Blue,OM643160). F-actin is stained with Omnimabs® 594-Phalloidin (Red,OM750007) diluted 1:100 for 30 minutes.
ELISA
Coat the plate with immunizing antigen.Blue line： Immune serum; Orange line: Pre-immune serum.| Application Notes | WB:1:1000-1:2000 ICC/IF:1:100-1:500 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C long term. Avoid freeze / thaw cycle. |
| Storage Buffer | PBS ONLY |
