Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-ITCH/AIP4 antibody [32/Itch] (ab280211)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Mouse monoclonal [32/Itch] to ITCH/AIP4
  • Suitable for: WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-ITCH/AIP4 antibody [32/Itch]
    See all ITCH/AIP4 primary antibodies

  • Description

    Mouse monoclonal [32/Itch] to ITCH/AIP4

  • Host species

    Mouse

  • Tested applications

    Suitable for: WBmore details
    Unsuitable for: ICC

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

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

  • Positive control

    • WB: Hela, Ramos, Raji, C6, PC-12, NIH/3T3, K562 whole cell lysates.

  • General notes

    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

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.

  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    32/Itch

  • Isotype

    IgG1

  • Research areas

Images

  • Western blot - Anti-ITCH/AIP4 antibody [32/Itch] (ab280211)
    Western blot - Anti-ITCH/AIP4 antibody [32/Itch] (ab280211)
    All lanes : Anti-ITCH/AIP4 antibody [32/Itch] (ab280211) at 1/1000 dilution

    Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
    Lane 2 : Ramos (human Burkitt's lymphoma B lymphocyte), whole cell lysate
    Lane 3 : Raji (human Burkitt's lymphoma B lymphocyte), whole cell lysate
    Lane 4 : C6 (rat glial tumor glial cell), whole cell lysate
    Lane 5 : PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate
    Lane 6 : NIH/3T3 (mouse embryonic fibroblast), whole cell lysate

    Lysates/proteins at 10 µg per lane.

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

    Predicted band size: 103 kDa
    Observed band size: 120 kDa
    why is the actual band size different from the predicted?



    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:11782481).

    The band around 75kDa is the degradation form. 50kDa band is the non-specific band according to KO validation.

    Exposure time Lane 1-2/4-5: 3 minutes Lane 3/6: 1 minute

  • Western blot - Anti-ITCH/AIP4 antibody [32/Itch] (ab280211)
    Western blot - Anti-ITCH/AIP4 antibody [32/Itch] (ab280211)
    All lanes : Anti-ITCH/AIP4 antibody [32/Itch] (ab280211) at 1/1000 dilution

    Lane 1 : Wild-type HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
    Lane 2 : ITCH knockout HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
    Lane 3 : K562 (human chronic myelogenous leukemia lymphoblast), whole cell lysate
    Lane 4 : Raji (human Burkitt's lymphoma B lymphocyte), 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: 103 kDa
    Observed band size: 120 kDa why is the actual band size different from the predicted?



    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 - ab280211 observed at 120 kDa. Red - loading control ab181602 (Rabbit monoclonal [EPR16891] to GAPDH) observed at 36 kDa.

    Lanes 1-2: ab280211 Anti-ITCH antibody was shown to react with ITCH in HeLa cells in Western blot. Loss of signal was observed when ITCH knockout sample was used. Wild-type and ITCH knockout samples were subjected to SDS-PAGE.

    ab280211 and Anti-GAPDH antibody [EPR16891] (ab181602) were incubated at 4? overnight at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680CW) (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800RD) (ab216772) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

    The band around 75kDa is the degradation form. 50kDa band is the non-specific band according to KO validation.

  • Anti-ITCH/AIP4 antibody [32/Itch] (ab280211)
    Anti-ITCH/AIP4 antibody [32/Itch] (ab280211)

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