SQSTM1 (p62) Antibody (C-term)

• Catalog Number: orb1931654
• Category: Antibodies
• Description: Purified Rabbit Polyclonal Antibody (Pab)
• Species/Host: Rabbit
• Clonality: Polyclonal
• Clone Number: RB4614
• Tested applications: IF, IHC-P, WB
• Predicted Reactivity: Rat
• Reactivity: Human, Mouse
• Isotype: Rabbit IgG
• Antibody Type: Primary Antibody
• Dilution range: IF: 1:200, IF: 1:50~100, WB: 1:2000, WB: 1:2000, IHC-P: 1:25, IHC-P: 1:50~100
• Form/Appearance: Purified polyclonal antibody supplied in PBS with 0.09% (W/V) sodium azide. This antibody is purified through a protein A column, followed by peptide affinity purification.
• Conjugation: Unconjugated
• MW: 47687 Da
• Target: This SQSTM1 (p62) antibody is generated from rabbits immunized with a KLH conjugated synthetic peptide between 317-346 amino acids of human SQSTM1 (p62).
• UniProt ID: Q13501
• NCBI: NP_001135771.1, NP_001135770.1, NP_003891.1
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles
• Alternative names: Sequestosome-1, EBI3-associated protein of 60 kDa,
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Staining SQSTM1 in Human prostate tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 3% BSA for 0.5 hour at room temperature; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody (1/25) for 1 hours at 37°C. A undiluted biotinylated goat polyvalent antibody was used as the secondary antibody.

Fluorescent image of U251 cells stained with SQSTM1 (p62) (C-term) antibody. U251 cells were treated with Chloroquine (50 μM, 16 h), then fixed with 4% PFA (20 min), permeabilized with Triton X-100 (0.2%, 30 min). Cells were then incubated with SQSTM1 (p62) (C-term) primary antibody (1:200, 2 h at room temperature). For secondary antibody, Alexa Fluor 488 conjugated donkey anti-rabbit antibody (green) was used (1:1000, 1 h). Nuclei were counterstained with Hoechst 33342 (blue) (10 μg/ml, 5 min). SQSTM1 (p62) immunoreactivity is localized to autophagic vacuoles in the cytoplasm of U251 cells.

Formalin-fixed and paraffin-embedded human cancer tissue reacted with the primary antibody, which was peroxidase-conjugated to the secondary antibody, followed by DAB staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated. BC = breast carcinoma; HC = hepatocarcinoma.

Immunofluorescence staining of Autophagy SQSTM1 (p62) Antibody (C-term) on Methanol-fixed and PFA fixed HeLa cells.

All lanes: Anti-SQSTM1 (p62) Antibody (C-term) at 1:2000 dilution. Lane 1: A549 whole cell lysate. Lane 2: C2C12 whole cell lysate. Lysates/proteins at 20 µg per lane. Secondary Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/10000 dilution. Predicted band size: 48 kDa. Blocking/Dilution buffer: 5% NFDM/TBST.

All lanes: Anti-SQSTM1 (p62) Antibody (C-term) at 1:2000 dilution. Lane 1: Jurkat whole cell lysate. Lane 2: THP-1 whole cell lysate. Lysates/proteins at 20 µg per lane. Secondary Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/10000 dilution. Predicted band size: 48 kDa. Blocking/Dilution buffer: 5% NFDM/TBST.