Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-SQSTM1 / p62 antibody [3/P62 LCK LIGAND] - BSA and Azide free (ab280091)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Mouse monoclonal [3/P62 LCK LIGAND] to SQSTM1 / p62 - BSA and Azide free
  • Suitable for: ICC, IP, IHC-P, WB
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-SQSTM1 / p62 antibody [3/P62 LCK LIGAND] - BSA and Azide free
    See all SQSTM1 / p62 primary antibodies

  • Description

    Mouse monoclonal [3/P62 LCK LIGAND] to SQSTM1 / p62 - BSA and Azide free

  • Host species

    Mouse

  • Tested applications

    Suitable for: ICC, IP, IHC-P, WBmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Wild-type HAP1; HeLa; MCF7; HEK-293 whole cell lysates. IHC-P: Human stomach carcinoma, lung carcinoma tissue. ICC: HeLa cells. IP: HeLa whole cell lysate.

  • General notes

    ab280091 is the carrier-free version of ab280086. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with

    ab280091 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

Properties

Images

  • Western blot - Anti-SQSTM1 / p62 antibody [3/P62 LCK LIGAND] - BSA and Azide free (ab280091)
    Western blot - Anti-SQSTM1 / p62 antibody [3/P62 LCK LIGAND] - BSA and Azide free (ab280091)
    All lanes : Anti-SQSTM1 / p62 antibody [3/P62 LCK LIGAND] (ab280086) at 1/1000 dilution

    Lane 1 : Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell line), whole cell lysate
    Lane 2 : SQSTM1 knockout HAP1 (human chronic myelogenous leukemia near-haploid cell line), whole cell lysate
    Lane 3 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
    Lane 4 : MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Mouse IgG H&L (IRDye® 800CW) (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) (ab216777) at 1/10000 dilution

    Predicted band size: 47 kDa
    Observed band size: 62 kDa
    why is the actual band size different from the predicted?



    This data was developed using ab280086, the same antibody clone in a different buffer formulation.

    Blocking and diluting buffer and concentration: Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS

    Lanes 1 - 4: Merged signal (red and green). Green - ab280086 observed at 62 kDa. Red - loading control ab181602 (Rabbit monoclonal [EPR16891] to GAPDH) observed at 36 kDa.

    Lanes 1-2: ab280086 Anti-SQSTM1/p62 antibody was shown to react with SQSTM1 in HAP1 cells in Western blot. Loss of signal was observed when SQSTM1 knockout sample was used. Wild-type and SQSTM1 knockout samples were subjected to SDS-PAGE. ab280086 and Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) were incubated at 4°C overnight at 1/1000 dilution and 1/20000 dilution respectively.

    Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680CW) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800RD) preadsorbed (ab216772) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging. 

  • Western blot - Anti-SQSTM1 / p62 antibody [3/P62 LCK LIGAND] - BSA and Azide free (ab280091)
    Western blot - Anti-SQSTM1 / p62 antibody [3/P62 LCK LIGAND] - BSA and Azide free (ab280091)
    All lanes : Anti-SQSTM1 / p62 antibody [3/P62 LCK LIGAND] (ab280086) at 1/1000 dilution

    Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell ), whole cell lysate
    Lane 2 : HEK-293 (human embryonic kidney epithelial cell), whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

    Predicted band size: 47 kDa
    Observed band size: 62 kDa why is the actual band size different from the predicted?



    This data was developed using ab280086, the same antibody clone in a different buffer formulation.

    Blocking and diluting buffer and concentration: 5% NFDM/TBST

    The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 24086455).

    Lysates were made freshly and used in WB immediately to minimize protein degradation.

    Exposure time: 15 seconds

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SQSTM1 / p62 antibody [3/P62 LCK LIGAND] - BSA and Azide free (ab280091)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SQSTM1 / p62 antibody [3/P62 LCK LIGAND] - BSA and Azide free (ab280091)

    This data was developed using ab280086 the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded Human stomach carcinoma tissue labeling SQSTM1 / p62 with ab280086 at 1/1000 dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human stomach carcinoma. The section was incubated with ab280086 for 30 mins at room temperature and followed by mouse specific IgG antibody (ab125913) for 8mins. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.

    Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SQSTM1 / p62 antibody [3/P62 LCK LIGAND] - BSA and Azide free (ab280091)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SQSTM1 / p62 antibody [3/P62 LCK LIGAND] - BSA and Azide free (ab280091)

    This data was developed using ab280086 the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded Human lung carcinoma tissue labeling SQSTM1 / p62 with ab280086 at 1/1000 dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human lung carcinoma. The section was incubated with ab280086 for 30 mins at room temperature and followed by mouse specific IgG antibody (ab125913) for 8mins. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.

    Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

  • Immunocytochemistry - Anti-SQSTM1 / p62 antibody [3/P62 LCK LIGAND] - BSA and Azide free (ab280091)
    Immunocytochemistry - Anti-SQSTM1 / p62 antibody [3/P62 LCK LIGAND] - BSA and Azide free (ab280091)

    This data was developed using ab280086 the same antibody clone in a different buffer formulation.

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa cells labelling SQSTM1 / p62 with ab280086 at 1/50 dilution, followed by ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in HeLa cell line. ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 dilution followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594)(Red). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

  • Immunoprecipitation - Anti-SQSTM1 / p62 antibody [3/P62 LCK LIGAND] - BSA and Azide free (ab280091)
    Immunoprecipitation - Anti-SQSTM1 / p62 antibody [3/P62 LCK LIGAND] - BSA and Azide free (ab280091)

    This data was developed using ab280086 the same antibody clone in a different buffer formulation.

    SQSTM1 / p62 was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell ), whole cell lysate 10 ug with ab280086 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab280086 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

    Lane 1: HeLa (human cervix adenocarcinoma epithelial cell ), whole cell lysate 10 ug

    Lane 2: ab280086 IP in HeLa whole cell lysate

    Lane 3: Mouse monoclonal IgG1(ab18443) instead of ab280086 in HeLa whole cell lysate

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 26 seconds

     

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