Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line, RAW 264.7 mouse monocyte/macrophage cell line, and C6 rat glioma cell line untreated (-) or treated (+) with 1 µg/mL LPS for 24 hours. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human/Mouse/Rat p62/SQSTM1 Monoclonal Antibody (Catalog # MAB8028) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for p62/SQSTM1 at approximately 62 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

p62/SQSTM1 in HeLa Human Cell Line.

p62/SQSTM1 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Mouse Anti-Human/Mouse/Rat p62/SQSTM1 Monoclonal Antibody (Catalog # MAB8028) at 25 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; NL007) and counterstained with DAPI (blue). Specific staining was localized to phagosomes in cell cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

p62/SQSTM1 in Human Liver.

p62/SQSTM1 was detected in immersion fixed paraffin-embedded sections of human liver using Mouse Anti-Human/Mouse/Rat p62/SQSTM1 Monoclonal Antibody (Catalog # MAB8028) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to nuclei and cytoplasm in hepatocytes. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Detection of Mouse p62/SQSTM1 by Simple Western^TM.

Simple Western lane view shows lysates of RAW 264.7 mouse monocyte/macrophage cell line untreated (-) or treated (+) with 1 µg/mL LPS for 24 hours, loaded at 0.2 mg/mL. A specific band was detected for p62/SQSTM1 at approximately 66 kDa (as indicated) using 20 µg/mL of Mouse Anti-Human/Mouse/Rat p62/SQSTM1 Monoclonal Antibody (Catalog # MAB8028). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

p62/SQSTM1 Specificity is Shown by Immunocytochemistry in Knockout Cell Line.

p62/SQSTM1 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line but is not detected in p62/SQSTM1 knockout (KO) HeLa cell line using Mouse Anti-Human/Mouse/Rat p62/SQSTM1 Monoclonal Antibody (Catalog # MAB8028) at 3 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Western Blot Shows Human p62/SQSTM1 Specificity Using Knockout Cell Line.

Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and p62/SQSTM1 knockout HeLa cell line (KO). PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human/Mouse/Rat p62/SQSTM1 Monoclonal Antibody (Catalog # MAB8028) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for p62/SQSTM1 at approximately 62 kDa (as indicated) in the parental HeLa cell line, but is not detectable in the knockout HeLa cell line. GAPDH (MAB5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Western Blot Shows Human p62/SQSTM1 Specificity Using Knockout Cell Line.

Western blot shows lysates of U2OS human osteosarcoma cell line and p62/SQSTM1 knockout U2OS cell line (KO). Nitrocellulose membrane was probed with 0.5 µg/mL of Mouse Anti-Human/Mouse/Rat p62/SQSTM1 Monoclonal Antibody (Catalog # MAB8028) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody. A specific band was detected for p62/SQSTM1 at approximately 62 kDa (as indicated) in the parental U2OS cell line, but is not detectable in knockout U2OS cell line. The Ponceau stained transfer of the blot is shown. This experiment was conducted under reducing conditions. Image, protocol, and testing courtesy of YCharOS Inc. See ycharos.com for additional details.

Detection of SQSTM1/p62 by Immunoprecipitation

Immunoprecipitation was performed on cell lysate of U2OS human osteosarcoma cell line using 1.0 μg of Mouse Anti-Human SQSTM1/p62 Monoclonal Antibody (Catalog # MAB8028) pre-coupled to protein G or protein A beads. Immunoprecipitated SQSTM1/p62 was detected with Rabbit Anti-SQSTM1/p62 Monoclonal Antibody (Catalog # MAB80281). The Ponceau stained transfers of each blot are shown. SM=10% starting material; UB=10% unbound fraction; IP=immunoprecipitated. Image, protocol, and testing courtesy of YCharOS Inc. (ycharos.com).

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Applications for Human/Mouse/Rat p62/SQSTM1 Antibody

Application

Recommended Usage

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed HeLa human cervical epithelial carcinoma cell line

Immunohistochemistry

5-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human liver

Immunoprecipitation

1 µg/1 mg cell lysate
Sample: Cell lysate of U2OS human osteosarcoma cell line

Knockout Validated

p62/SQSTM1 is specifically detected in HeLa human cervical epithelial carcinoma parental cell line and parental U2OS cell line but is not detectable in p62/SQSTM1 knockout HeLa cell line and knockout U2OS cell line.

Simple Western

20 µg/mL
Sample: RAW 264.7 mouse monocyte/macrophage cell line treated with LPS

Western Blot

2 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line, RAW 264.7 mouse monocyte/macrophage cell line, and C6 rat glioma cell line untreated or treated with LPS

Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 2 reviews rated 5 using MAB8028 in the following applications:

Immunocytochemistry/Immunofluorescence (1 Review)
Western Blot (1 Review)

Formulation, Preparation, and Storage

Reconstitution

Sterile PBS to a final concentration of 0.5 mg/mL. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.
*Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Stability & Storage

Store the unopened product at -20 to -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Do not use past expiration date.

Background: p62/SQSTM1

SQSTM1 (Sequestrome-1), also called p62, is a widely expressed, stress-inducible, multifunctional 62 kDa intracellular protein. The 440 amino acid (aa) human SQSTM1 contains multiple adaptor domains that allow interaction with proteins in NGF/NFkB and other signaling pathways (notably TRAF6, atypical protein kinase C family and Src family), polyubiquitin, proteasome subunits and many others. It contains numerous regulatory phosphorylation sites and a dimerization site. SQSTM1 shuttles ubiquitinylated proteins to the proteasome and is important in autophagy and apoptosis. Its dysregulation is associated with Paget’s disease of bone, Parkinson’s and Alzheimer’s diseases, and cancers. Within aa 344-440, which includes the ubiquitin-binding domain, human SQSTM1 shares 100% aa sequence identity with mouse and rat SQSTM1.

Long Name

Sequestosome 1

Alternate Names

A170, EBIAP, ORCA, OSIL, PDB3, Sequestosome 1, SQSTM1, ZIP3

Entrez Gene IDs

8878 (Human); 18412 (Mouse); 113894 (Rat)

Gene Symbol

SQSTM1

UniProt

Q13501

Additional p62/SQSTM1 Products

Product Documents for Human/Mouse/Rat p62/SQSTM1 Antibody

Product Datasheets

Download Product Datasheets

COA

SDS

Download SDS

Product Specific Notices for Human/Mouse/Rat p62/SQSTM1 Antibody

For research use only

Citations
Reviews

Human/Mouse/Rat p62/SQSTM1 Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB8028

Key Product Details

Validated by

Species Reactivity

Validated:

Cited:

Applications

Validated:

Cited:

Label

Antibody Source

View all p62/SQSTM1 Antibodies »

Product Summary for Human/Mouse/Rat p62/SQSTM1 Antibody

Specificity

Clonality

Host

Isotype

Scientific Data Images for Human/Mouse/Rat p62/SQSTM1 Antibody

Detection of Human, Mouse, and Rat p62/SQSTM1 by Western Blot.

p62/SQSTM1 in HeLa Human Cell Line.

p62/SQSTM1 in Human Liver.

Detection of Mouse p62/SQSTM1 by Simple WesternTM.

p62/SQSTM1 Specificity is Shown by Immunocytochemistry in Knockout Cell Line.

Western Blot Shows Human p62/SQSTM1 Specificity Using Knockout Cell Line.

Western Blot Shows Human p62/SQSTM1 Specificity Using Knockout Cell Line.

Detection of SQSTM1/p62 by Immunoprecipitation

Applications for Human/Mouse/Rat p62/SQSTM1 Antibody

Immunocytochemistry

Immunohistochemistry

Immunoprecipitation

Knockout Validated

Simple Western

Western Blot

Reviewed Applications

Formulation, Preparation, and Storage

Reconstitution

Formulation

Stability & Storage

Background: p62/SQSTM1

Long Name

Alternate Names

Entrez Gene IDs

Gene Symbol

UniProt

Additional p62/SQSTM1 Products

Product Documents for Human/Mouse/Rat p62/SQSTM1 Antibody

Product Datasheets

COA

Certificate of Analysis

SDS

Product Specific Notices for Human/Mouse/Rat p62/SQSTM1 Antibody

Detection of Mouse p62/SQSTM1 by Simple Western^TM.