Key Product Details

Validated by

Knockout/Knockdown, Biological Validation

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human

Applications

Validated:

Knockout Validated, Western Blot, Immunocytochemistry, Immunoprecipitation

Cited:

Western Blot, Immunocytochemistry, Immunoprecipitation

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Mouse IgG2B Clone # 864807R
View all p62/SQSTM1 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human p62/SQSTM1
Asp368-Leu440
Accession # Q13501

Specificity

Detects human p62/SQSTM1 in ELISAs. Detects human, mouse and rat p62/SQSTM1 in Western blots

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2B

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

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

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

Western blot shows lysates of A172 human glioblastoma cell line and RAW 264.7 mouse monocyte/macrophage cell line untreated (-) or treated (+) with 10 µg/mL LPS for 4 hours. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human/Mouse/Rat p62/SQSTM1 Monoclonal Antibody (Catalog # MAB8028R) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # 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 antibody in HeLa Human Cell Line by Immunocytochemistry (ICC).

p62/SQSTM1 in HeLa Human Cell Line.

p62/SQSTM1 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line treated with staurosporine using Mouse Anti-Human/Mouse/Rat p62/SQSTM1 Monoclonal Antibody (Catalog # MAB8028R) at 3 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # 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 Antibody Specificity Using Knockout Cell Line.

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 1 µg/mL of Mouse Anti-Human/Mouse/Rat p62/SQSTM1 Monoclonal Antibody (Catalog # MAB8028) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # 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 (Catalog # 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 # MAB8028R) 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 # MAB8028R) 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).

Applications for Human/Mouse/Rat p62/SQSTM1 Antibody

Application
Recommended Usage

Immunocytochemistry

3-25 µg/mL
Sample: Immersion fixed HeLa human cervical epithelial carcinoma cell line treated with staurosporine

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. 

Western Blot

1 µg/mL
Sample: A172 human glioblastoma cell line and RAW 264.7 mouse monocyte/macrophage cell line treated with LPS

Formulation, Preparation, and Storage

Purification

Protein A or G purified from cell culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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

For research use only

Related Research Areas

Autophagy
NFkB Pathway

Citations for Human/Mouse/Rat p62/SQSTM1 Antibody

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Protocols

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