WB result of SQSTM1/p62 Rabbit mAb
Primary antibody: SQSTM1/p62 Rabbit mAb at 1/1000 dilution
Lane 1: HeLa whole cell lysate 20 µg
Lane 2: Jurkat whole cell lysate 20 µg
Lane 3: Hep G2 whole cell lysate 20 µg
Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 48 kDa
Observed MW: 62 kDa
(This blot was developed with high sensitivity substrate)
SQSTM1/p62 Recombinant Rabbit mAb (S-778-1)
SQSTM1/p62 Recombinant Rabbit mAb (S-778-1)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Synonyms | Sequestosome-1, EBI3-associated protein of 60 kDa (EBIAP, p60), Phosphotyrosine-independent ligand for the Lck SH2 domain of 62 kDa, Ubiquitin-binding protein p62, ORCA, OSIL |
| Immunogen | Synthetic Peptide |
| Location | Cytoplasm |
| Accession | Q13501 |
| Clone Number | S-778-1 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, IHC-P, ICC, ICFCM, IP |
| Reactivity | Hu, Ms, Rt |
| Predicted Reactivity | Or |
| Purification | Protein A |
| Concentration | 0.5 mg/ml |
| Conjugation | Unconjugated |
| Physical Appearance | Liquid |
| Storage Buffer | PBS, 40% Glycerol, 0.05% BSA, 0.03% Proclin 300 |
| Stability & Storage | 12 months from date of receipt / reconstitution, -20 °C as supplied. |
Dilution
| application | dilution | species |
| WB | 1:1000 | |
| IP | 1:50 | |
| IHC-P | 1:500 | |
| ICC | 1:500 | |
| ICFCM | 1:500 |
Background
Sequestosome-1 is a protein that in humans is encoded by the SQSTM1 gene. Also known as the ubiquitin-binding protein p62, it is an autophagosome cargo protein that targets other proteins that bind to it for selective autophagy. By interacting with GATA4 and targeting it for degradation, it can inhibit GATA-4 associated senescence and senescence-associated secretory phenotype. Mutations in SQSTM1 are a common cause of Paget's disease of bone.
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Picture
Western Blot
WB result of SQSTM1/p62 Rabbit mAb
Primary antibody: SQSTM1/p62 Rabbit mAb at 1/1000 dilution
Lane 1: mouse brain lysate 20 µg
Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 48 kDa
Observed MW: 62 kDa
(This blot was developed with high sensitivity substrate)
WB result of SQSTM1/p62 Rabbit mAb
Primary antibody: SQSTM1/p62 Rabbit mAb at 1/1000 dilution
Lane 1: rat brain lysate 20 µg
Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 48 kDa
Observed MW: 62 kDa
(This blot was developed with high sensitivity substrate)
FC
Flow cytometric analysis of 4% PFA fixed 90% methanol permeabilized HepG2 (Human hepatocellular carcinoma epithelial cell) cells labelling SQSTM1/p62 antibody at 1/500 dilution (0.1 μg)/ (Red) compared with a Rabbit monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti - Rabbit IgG Alexa Fluor® 488 was used as the secondary antibody.
IP
SQSTM1/p62 Rabbit mAb at 1/50 dilution (1 µg) immunoprecipitating SQSTM1/p62 in 0.4 mg HeLa whole cell lysate.
Western blot was performed on the immunoprecipitate using SQSTM1/p62 Rabbit mAb at 1/1000 dilution.
Secondary antibody (HRP) for IP was used at 1/400 dilution.
Lane 1: HeLa whole cell lysate 20 µg (Input)
Lane 2: SQSTM1/p62 Rabbit mAb IP in HeLa whole cell lysate
Lane 3: Rabbit monoclonal IgG IP in HeLa whole cell lysate
Predicted MW: 48 kDa
Observed MW: 62 kDa
Immunohistochemistry
IHC shows positive staining in paraffin-embedded human colon. Anti-SQSTM1/p62 antibody was used at 1/500 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human colon cancer. Anti-SQSTM1/p62 antibody was used at 1/500 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human lung squamous cell carcinoma. Anti-SQSTM1/p62 antibody was used at 1/500 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human testis. Anti-SQSTM1/p62 antibody was used at 1/500 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded mouse cerebral cortex. Anti-SQSTM1/p62 antibody was used at 1/500 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded rat liver. Anti-SQSTM1/p62 antibody was used at 1/500 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
Immunocytochemistry
ICC analysis of HeLa cells treated with Chloroquine (50μM, 24 hours) (top panel) and HeLa cells untreated with Chloroquine (50μM, 24 hours) (below panel). Anti-SQSTM1 antibody was used at 1/500 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 100% ice-cold methanol and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (Red).
