SQSTM1 Antibody

• Catalog Number: orb1240011
• Category: Antibodies
• Description: SQSTM1 Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, IF, IHC-P, WB
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: SQSTM1 antibody was raised against a 13 amino acid synthetic peptide from near the carboxy terminus of human SQSTM1.The immunogen is located within the last 50 amino acids of SQSTM1.
• Antibody Type: Primary Antibody
• Concentration: 1 mg/mL
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: Predicted: 48kDObserved: 65 kD
• Target: SQSTM1
• UniProt ID: Q13501
• NCBI: Q13501
• 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.
• Buffer/Preservatives: SQSTM1 Antibody is supplied in PBS containing 0.02% sodium azide.
• Alternative names: SQSTM1 Antibody: p60, p62, A170, OSIL, PDB3, ZIP3,
• Note: For research use only
• Application notes: WB: 0.5-2 μg/mL; IF: 20 μg/mL; IHC: 2-5 μg/mL;.Antibody validated: Western Blot in human and mouse samples; Immunofluorescence and Immunohistochemistry in human, mouse and rat samples. All other applications and species not yet tested.
• Expiration Date: 12 months from date of receipt.

Immunofluorescence Validation of SQSTM1 In HeLa Cells. Immunofluorescent analysis of PFA-fixed HeLa cells labeling SQSTM1 with orb1240011 at 20 µg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/1000 dilution (red) and Hoechst staining (blue). Alpha tubulin was stained with anti-alpha tubulin antibody following by goat anti-mouse IgG secondary antibody (green). Images were captured with confocal microscopy.

Immunofluorescence Validation of SQSTM1 in Mouse Spleen Tissue. Immunofluorescent analysis of 4% paraformaldehyde-fixed Mouse Spleen Tissue labeling SQSTM1 with orb1240011 at 20 µg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).

Immunohistochemistry Validation of SQSTM1 in Human Spleen Tissue. Immunohistochemical analysis of paraffin-emb edded Human Spleen Tissue using anti-SQSTM1 antibody (orb1240011) at 2 µg/mL. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

Immunohistochemistry Validation of SQSTM1 in Mouse Spleen Tissue. Immunohistochemical analysis of paraffin-embedded Mouse Spleen Tissue using anti-SQSTM1 antibody (orb1240011) at 2 µg/mL. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

Immunohistochemistry Validation of SQSTM1 in Rat Spleen Tissue. Immunohistochemical analysis of paraffin-embedded Rat Spleen Tissue using anti-SQSTM1 antibody (orb1240011) at 2 µg/mL. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

Induced Expression Validation in Mouse Macrophage Cells. Loading: 15 µg of lysates per lane. Antibodies: SQSTM1 orb1240011 (0.5 µg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. Raw 264.7 cells were treated with LPS (0.3 µg /mL) for different time period (0-24 hrs). There was an increase in SQSTM1 protein expression overtime after LPS treatment.

KO Validation in HEK293T Cells. Loading: 10 µg of HEK293T WT cell lysates or SQSTM1 KO cell lysates. Antibodies: SQSTM1 orb1240011 (1 µg/mL) and beta-actin orb1240312 (1 µg/mL), 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.

Western Blot Validation in Cell Lines. Loading: 15 µg of lysates per lane. Antibodies: SQSTM1 orb1240011 (0.5 µg/mL), 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.

Western Blot Validation in Mouse Tissues. Loading: 15 µg of lysates per lane. Antibodies: SQSTM1 orb1240011 (1 µg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.

Western Blot Validation in Rat Tissues. Loading: 15 µg of lysates per lane. Antibodies: SQSTM1 orb1240011 (1 µg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.