Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-CD58 antibody [EP15041] (ab196648)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP15041] to CD58
  • Suitable for: WB
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-CD58 antibody [EP15041]
    See all CD58 primary antibodies

  • Description

    Rabbit monoclonal [EP15041] to CD58

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    WB
    Human
    See all applications and species data

  • Immunogen

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

  • Positive control

    • WB: HeLa, Hap1, Daudi, Raji, Jurkat, TF-1, SW480 and K562 cell lysates.

  • General notes

     

     

    Previously labelled as LFA3.

    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: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EP15041

  • Isotype

    IgG

  • Research areas

Images

  • Western blot - Anti-CD58 antibody [EP15041] (ab196648)
    Western blot - Anti-CD58 antibody [EP15041] (ab196648)
    All lanes : Anti-CD58 antibody [EP15041] (ab196648) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : CD58 knockout HeLa cell lysate
    Lane 3 : HAP1 cell lysate
    Lane 4 : Daudi cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 28 kDa



    Lanes 1-4: Merged signal (red and green). Green - ab196648 observed at 43 kDa. Red - loading control ab8245 observed at 37 kDa.

     ab196648 Anti-CD58 antibody [EP15041] was shown to specifically react with CD58 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265947 (knockout cell lysate ab257880) was used. Wild-type and CD58 knockout samples were subjected to SDS-PAGE. ab196648 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 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-CD58 antibody [EP15041] (ab196648)
    Western blot - Anti-CD58 antibody [EP15041] (ab196648)
    All lanes : Anti-CD58 antibody [EP15041] (ab196648) at 1/1000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : CD58 knockout HAP1 whole cell lysate

    Lysates/proteins at 40 µg per lane.

    Predicted band size: 28 kDa
    Observed band size: 43 kDa
    why is the actual band size different from the predicted?



    Lanes 1 - 4: Merged signal (red and green). Green - ab196648 observed at 43 kDa. Red - loading control, ab130007, observed at 130 kDa.

     ab196648 was shown to specifically react with CD58 in wild-type HAP1 cells as signal was lost in CD58 knockout cells. Wild-type and CD58 knockout samples were subjected to SDS-PAGE. Ab196648 and ab130007 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-CD58 antibody [EP15041] (ab196648)
    Western blot - Anti-CD58 antibody [EP15041] (ab196648)
    All lanes : Anti-CD58 antibody [EP15041] (ab196648) at 1/5000 dilution

    Lane 1 : Raji (Human Burkitt's lymphoma cell line) whole cell lysate
    Lane 2 : Jurkat (Human T cell leukemia cells from peripheral blood) whole 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: 28 kDa
    Observed band size: 65-70 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-CD58 antibody [EP15041] (ab196648)
    Western blot - Anti-CD58 antibody [EP15041] (ab196648)
    All lanes : Anti-CD58 antibody [EP15041] (ab196648) at 1/1000 dilution

    Lane 1 : TF-1 (Human bone marrow erythroleukemia cells) whole cell lysate
    Lane 2 : K562 (Human chronic myelogenous leukemia cells from bone marrow) whole 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: 28 kDa
    Observed band size: 65-70 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-CD58 antibody [EP15041] (ab196648)
    Western blot - Anti-CD58 antibody [EP15041] (ab196648)
    All lanes : Anti-CD58 antibody [EP15041] (ab196648) at 1/2000 dilution

    Lane 1 : SW480 (Human colorectal adenocarcinoma cell line) control
    Lane 2 : SW480 (Human colorectal adenocarcinoma cell line) de-glycosylation

    Lysates/proteins at 10 µg per lane.

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

    Predicted band size: 28 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Observed band size : 65-70 (27, 28) kDa.

  • Anti-CD58 antibody [EP15041] (ab196648)
    Anti-CD58 antibody [EP15041] (ab196648)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

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