Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-SIKE1 antibody [EPR14692] (ab183509)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR14692] to SIKE1
  • Suitable for: IP, IHC-P, WB
  • Knockout validated
  • Reacts with: Mouse, Human

Overview

  • Product name

    Anti-SIKE1 antibody [EPR14692]
    See all SIKE1 primary antibodies

  • Description

    Rabbit monoclonal [EPR14692] to SIKE1

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    IHC-P
    Human
    IP
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

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

  • Positive control

    • SH-SY5Y, HeLa and HEK-293T cell lysates; Human kidney tissue.

  • 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.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

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

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol (glycerin, glycerine), 59% PBS, 0.05% BSA
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR14692

  • Isotype

    IgG

  • Research areas

Images

  • Western blot - Anti-SIKE1 antibody [EPR14692] (ab183509)
    Western blot - Anti-SIKE1 antibody [EPR14692] (ab183509)
    All lanes : Anti-SIKE1 antibody [EPR14692] (ab183509) at 1/10000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : Suppressor of IKBKE 1 knockout HeLa cell lysate
    Lane 3 : HEK-293 cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 24 kDa



    Lanes 1-3: Merged signal (red and green). Green - ab183509 observed at 25 kDa. Red - loading control ab8245 observed at 37 kDa.

     ab183509 Rabbit monoclonal [EPR14692] to SIKE1 was shown to specifically react with Suppressor of IKBKE 1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265933 (knockout cell lysate ab258188) was used. Wild-type and Suppressor of IKBKE 1 knockout samples were subjected to SDS-PAGE. ab183509 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 10000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

     

  • Western blot - Anti-SIKE1 antibody [EPR14692] (ab183509)
    Western blot - Anti-SIKE1 antibody [EPR14692] (ab183509)
    All lanes : Anti-SIKE1 antibody [EPR14692] (ab183509) at 1/10000 dilution

    Lane 1 : Wild-type HAP1 cell lysate
    Lane 2 : SIKE1 knockout HAP1 cell lysate
    Lane 3 : HeLa cell lysate
    Lane 4 : PC3 cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 24 kDa



    Lanes 1 - 4: Merged signal (red and green). Green - ab183509 observed at 25 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab183509 was shown to specifically react with SIKE1 when SIKE1 knockout samples were used. Wild-type and SIKE1 knockout samples were subjected to SDS-PAGE. ab183509 and ab8245 (loading control to GAPDH) were both diluted 1/10 000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-SIKE1 antibody [EPR14692] (ab183509)
    Western blot - Anti-SIKE1 antibody [EPR14692] (ab183509)
    All lanes : Anti-SIKE1 antibody [EPR14692] (ab183509) at 1/20000 dilution

    Lane 1 : SH-SY5Y cell lysate
    Lane 2 : Hela cell lysate
    Lane 3 : 293 cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 24 kDa
    Additional bands at: 24 kDa. We are unsure as to the identity of these extra bands.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIKE1 antibody [EPR14692] (ab183509)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIKE1 antibody [EPR14692] (ab183509)

    Immunohistochemical analysis of paraffin embedded Human kidney tissue labeling SIKE1 with ab183509 at 1/50.

    Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

  • Immunoprecipitation - Anti-SIKE1 antibody [EPR14692] (ab183509)
    Immunoprecipitation - Anti-SIKE1 antibody [EPR14692] (ab183509)

    Lysate from 293 cells was immunoprecipitated with ab183509 at a 1/50 dilution. For the subsequent blot, ab183509 used at a 1/10000 dilution with an anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at a 1/1500 dilution for the secondary. Blocking/ Dilution buffer: 5% NFDM/TBST. Lane 2: TBS instead of lysate.

  • Anti-SIKE1 antibody [EPR14692] (ab183509)
    Anti-SIKE1 antibody [EPR14692] (ab183509)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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