Anti-SQSTM1/p62 Picoband™ Antibody


SKU PB9444
Size 100μg/vial
Reactivity Human, Monkey, Mouse, Rat
Clonality Polyclonal
Host Rabbit
Ig Isotype N/A
Applications IF, IHC, ICC, WB

Overview

Product Name Anti-SQSTM1/p62 Picoband™ Antibody
SKU/Catalog Number PB9444
Storage & Handling At -20°C for one year. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for a longer time.Avoid repeated freezing and thawing.
Size 100μg/vial
Description Rabbit IgG polyclonal antibody for Sequestosome-1(SQSTM1) detection. Tested with WB, IHC-P, IHC-F, ICC, IF in Human;Mouse;Monkey;Rat.
Cite This Product Anti-SQSTM1/p62 Picoband™ Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9444)
Host Rabbit
Contents/Buffer Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Form Lyophilized
Immunogen A synthetic peptide corresponding to a sequence at the N-terminus of human SQSTM1/p62 (69-96aa DEDGDLVAFSSDEELTMAMSYVKDDIFR), identical to the related mouse and rat sequences.
Reactivity Human, Monkey, Mouse, Rat

Assay Details

Assay Dilutions Overview

Concentration: Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Immunohistochemistry(Paraffin-embedded Section), 0.5-1μg/ml, Human, Mouse, Rat, By Heat
Western blot, 0.1-0.5μg/ml, Human, Mouse, Monkey, Rat
Immunofluorescence, 2μg/ml, Human
Immunohistochemistry(Frozen Section), 0.5-1μg/ml, Mouse, Rat
Immunocytochemistry, 0.5-1μg/ml, Human

Boster's Secondary Antibodies And IHC, WB Kits

The following reagents are used to generate the images below.

Boster recommends Enhanced Chemiluminescent Kit with anti-Rabbit IgG (EK1002) for Western blot, and HRP Conjugated anti-Rabbit IgG Super Vision Assay Kit (SV0002-1) for IHC(P), IHC(F) and ICC.

Images And Assay Conditions

Figure 4. IF analysis of SQSTM1 using anti- SQSTM1 antibody (PB9444).
SQSTM1 was detected in immunocytochemical section of A431 cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL rabbit anti- SQSTM1 Antibody (PB9444) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Figure 1. IHC analysis of SQSTM1 using anti-SQSTM1 antibody (PB9444).
SQSTM1 was detected in paraffin-embedded section of human intestinal cancer tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-SQSTM1 Antibody (PB9444) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

Figure 2. IHC analysis of SQSTM1 using anti-SQSTM1 antibody (PB9444).
SQSTM1 was detected in paraffin-embedded section of mouse small intestine tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-SQSTM1 Antibody (PB9444) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

Figure 3. IHC analysis of SQSTM1 using anti-SQSTM1 antibody (PB9444).
SQSTM1 was detected in immunocytochemical section of A549 cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 1μg/ml rabbit anti-SQSTM1 Antibody (PB9444) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

Figure 5. IF analysis of SQSTM1/p62 using anti- SQSTM1/p62 antibody (PB9444)
SQSTM1/p62 was detected in paraffin-embedded section of human lung cancer tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution ) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/mL rabbit anti- SQSTM1/p62 Antibody (PB9444) overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Figure 6. Western blot analysis of SQSTM1 using anti-SQSTM1 antibody (PB9444).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: human placenta tissue lysates,
Lane 2: COS7 whole cell lysates
Lane 3: A549 whole cell lysates,
Lane 4: U87 whole cell lysates.
Lane 5: Hela whole cell lysates.
Lane 6: Caco-2 whole cell lysates.
Lane 7: Hepg2 whole cell lysates.
Lane 8: THP-1 whole cell lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-SQSTM1 antigen affinity purified polyclonal antibody (Catalog # PB9444) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for SQSTM1 at approximately 62KD. The expected band size for SQSTM1 is at 62KD.

Figure 7. Western blot analysis of SQSTM1 using anti-SQSTM1 antibody (PB9444).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: rat PC-12 whole cell lysates
Lane 2: rat RH35 whole cell lysates
Lane 3: mouse HEPA1-6 whole cell lysates
Lane 4: mouse NIH/3T3 whole cell lysates
Lane 5: mouse RAW246.7 whole cell lysates
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-SQSTM1 antigen affinity purified polyclonal antibody (Catalog # PB9444) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for SQSTM1 at approximately 62KD. The expected band size for SQSTM1 is at 62KD.

Figure 8. IHC analysis of SQSTM1 using anti-SQSTM1 antibody (PB9444).
SQSTM1 was detected in frozen section of human placenta tissue. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-SQSTM1 Antibody (PB9444) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

Figure 9. IHC analysis of SQSTM1 using anti-SQSTM1 antibody (PB9444).
SQSTM1 was detected in paraffin-embedded section of rat small intestine tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-SQSTM1 Antibody (PB9444) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

Target Info

Protein Target Info (Source: Uniprot.org)

Uniprot Id Q13501
Gene Name SQSTM1
Protein Name Sequestosome-1
Tissue Specificity Ubiquitously expressed. .
Alternative Names 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;SQSTM1;ORCA, OSIL;
Subcellular Localization Cytoplasm. Late endosome. Lysosome. Cytoplasmic vesicle, autophagosome. Nucleus. Endoplasmic reticulum. Cytoplasm, P-body. Sarcomere (By similarity). In cardiac muscles localizes to the sarcomeric band (By similarity). Commonly found in inclusion bodies containing polyubiquitinated protein aggregates. In neurodegenerative diseases, detected in Lewy bodies in Parkinson disease, neurofibrillary tangles in Alzheimer disease, and HTT aggregates in Huntington disease. In protein aggregate diseases of the liver, found in large amounts in Mallory bodies of alcoholic and nonalcoholic steatohepatitis, hyaline bodies in hepatocellular carcinoma, and in SERPINA1 aggregates. Enriched in Rosenthal fibers of pilocytic astrocytoma. In the cytoplasm, observed in both membrane-free ubiquitin- containing protein aggregates (sequestosomes) and membrane- surrounded autophagosomes. Colocalizes with TRIM13 in the perinuclear endoplasmic reticulum. Co-localizes with TRIM5 in the cytoplasmic bodies. .
Molecular Weight 47687 MW

*if product is indicated to react with multiple species, protein info is based on the human gene.

Ontology

Protein Function Autophagy receptor that interacts directly with both the cargo to become degraded and an autophagy modifier of the MAP1 LC3 family. Required both for the formation and autophagic degradation of polyubiquitin-containing bodies, called ALIS (aggresome-like induced structures) and links ALIS to the autophagic machinery. Involved in midbody ring degradation. May regulate the activation of NFKB1 by TNF-alpha, nerve growth factor (NGF) and interleukin- 1. May play a role in titin/TTN downstream signaling in muscle cells. May regulate signaling cascades through ubiquitination. Adapter that mediates the interaction between TRAF6 and CYLD (By similarity). May be involved in cell differentiation, apoptosis, immune response and regulation of K(+) channels. .
Research Areas Human, Monkey, Mouse, Rat

*You can search these to find other products in these research areas.
Background SQSTM1(Sequestosome-1), also known as Ubiquitin-Binding Protein P62 or P62, is a protein that in humans is encoded by the SQSTM1 gene. The Src homology type 2 (SH2) domain is a highly conserved motif of about 100 amino acids which mediates protein-protein interactions by binding to phosphotyrosine.p56-lck, a T-cell-specific src family tyrosine kinase with an SH2 domain, is involved in T-cell signal transduction. The International Radiation Hybrid Mapping Consortium mapped the p62 gene to chromosome 5q35. Park et al. (1995) found that the p56-lck SH2 domain binds to p62 at the ser59 of p62 only when that serine is phosphorylated. Joung et al. (1996) expressed epitope-tagged p62 in Hela cells and showed that the expressed protein bound to the lck SH2 domain and that this binding was dependent on the N-terminal 50 amino acids of p62 but not on the tyrosine residue in this region.

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Polyclonal antibody for p62/SQSTM1 detection. Host: Rabbit.Size: 100μg/vial. Tested applications: IHC-P. Reactive species: Human. p62/SQSTM1 information: Molecular Weight: 47687 MW; Subcellular Localization: Cytoplasm. Late endosome. Lysosome. Cytoplasmic vesicle, autophagosome. Nucleus. Endoplasmic reticulum. Cytoplasm, P-body. Sarcomere (By similarity). In cardiac muscles localizes to the sarcomeric band (By similarity). Commonly found in inclusion bodies containing polyubiquitinated protein aggregates. In neurodegenerative diseases, detected in Lewy bodies in Parkinson disease, neurofibrillary tangles in Alzheimer disease, and HTT aggregates in Huntington disease. In protein aggregate diseases of the liver, found in large amounts in Mallory bodies of alcoholic and nonalcoholic steatohepatitis, hyaline bodies in hepatocellular carcinoma, and in SERPINA1 aggregates. Enriched in Rosenthal fibers of pilocytic astrocytoma. In the cytoplasm, observed in both membrane-free ubiquitin- containing protein aggregates (sequestosomes) and membrane- surrounded autophagosomes. Colocalizes with TRIM13 in the perinuclear endoplasmic reticulum. Co-localizes with TRIM5 in the cytoplasmic bodies; Tissue Specificity: Ubiquitously expressed.
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In stock
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PB9444
Buy one primary antibody get one 0.5ml HRP or Biotin secondary antibody for free.
*Sample sizes are prepared on demand and will take extra lead time. (cannot be conjugated)
$280.00

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Publications

Regulation of autophagy by AMP-activated protein kinase/sirtuin 1 pathway reduces spinal cord neurons damage
Interferon regulatory factor-1 activates autophagy to aggravate hepatic ischemia-reperfusion injury via the P38/P62 pathway in mice
Nrf2 signalling andautophagyare involved in diabetes mellitus-induced defects in the development of mouse placenta
Dual attenuation of proteasomal and autophagic BMAL1 degradation in Clock?19/+ mice contributes to improved glucose homeostasis
Disturbed Flow Induces Autophagy, but Impairs Autophagic Flux to Perturb Mitochondrial Homeostasis
Song J, Hu Y, Li J, Zheng H, Wang J, Guo L, Shi H, Liu L. Arch Virol. 2018 Jan;163(1):135-144. doi: 10.1007/s00705-017-3592-x. Epub 2017 Oct 19. Suppression of the toll-like receptor 7-dependent type I interferon production pathway by autophagy re...

Customer Q&As

  • Q: Do you offer BSA-free antibodies? Keyword: Bovine serum albumin, carrier protein, conjugation
    A: Yes, please contact us at support@bosterbio.com for more information about BSA-free antibodies and availability. The new BSA-free formula uses trehalose as a replacement to BSA. We have tested many alternative chemicals and found that trehalose protects the antibodies the best.
  • Q: Can I conjugate markers to this antibody? Can I link custom conjugates to this antibody? Keyword: conjugation
    A: The antibody is stored with BSA and cannot be conjugated with markers. Carrier free antibodies are available upon request. Please contact support@bosterbio.com
  • Q: What should I use for negative control?
    A: Please contact us for negative control suggestions. You can also check expression databases such as genecards, uniprot etc. Due to logistic reasons, we do not sell serum or lysates that we use internally for positive or negative control.
  • Q: Where can I find troubleshooting information? What should I do if I have unexpected bands, high background, no signal, weak signal
    A: You can find Boster's troubleshoot guides under tech support tab. Please contact us for further assistance on troubleshooting your experiment.
  • Q: What is the immunogen sequence of this antibody? Is this antibody polyclonal or monoclonal?
    A: You can find the immunogen sequence under "
  • Q: What is the expected band size? Why is it different than the observed band size?
    A: The expected band size is predicted on the size of the protein. The actual band size may be affected by a few other factors including but not limited to:<br>1. Post-translational modification:phosphorylation, methylation, glycosylation etc. These modifications prevent SDS molecules from binding to the target protein and thus make the band size appear larger than expected<br>2. Post-translational cleavage: this can cause smaller bands and or multiple bands <br><br>3. Alternative splicing: the same gene can have alternative splicing patterns generating different size proteins, all with reactivities to the antibody. <br><br>4. Amino Acid R chain charge: SDS binds to positive charges. The different size and charge of the Amino Acid side chains can affect the amount of SDS binding and thus affect the observed band size.<br>5. Multimers: Multimers are usually broken up in reducing conditions. However if the interactions between the multimers are strong, the band may appear higher., <br>
  • Q: What is the suggested dilution ratio for Western Blot (WB), Immunohistochemistry (IHC) and or ELISA standards? What is the optimal pH for the sample?
    A: Check the datasheet for the product for details on dilution ratios for different experiments. You can find the datasheet button on the right side of the product page.
  • Q: What is the protocol you used for your Western blotting (WB) and Immunohistochemistry (IHC)?
    A: Check our protocols under the tech support tab.
  • Q: What are some alternative names that could be used to describe this product?
    A: Some common names include but are not limited to p62 antibody, sequestosome 1 antibody, sequestosome-1 antibody, sqstm1 antibody
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