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  1. Link

    ly-6k-antibody-epr23021-61-ab246486.pdf

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Immunology Adaptive Immunity B Cells Non-CD
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Ly-6K antibody [EPR23021-61] (ab246486)

  • Datasheet
  • SDS
  • Certificate of Compliance
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ly-6K antibody [EPR23021-61] (ab246486)
  • Western blot - Anti-Ly-6K antibody [EPR23021-61] (ab246486)
  • Western blot - Anti-Ly-6K antibody [EPR23021-61] (ab246486)
  • Immunoprecipitation - Anti-Ly-6K antibody [EPR23021-61] (ab246486)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ly-6K antibody [EPR23021-61] (ab246486)
  • Anti-Ly-6K antibody [EPR23021-61] (ab246486)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR23021-61] to Ly-6K
  • Suitable for: WB, IHC-P, IP
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

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Secondary
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Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
Knockout
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Human LY6K (Ly-6K) knockout HeLa cell line (ab265874)
Primary
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Anti-Ly-6K antibody [EPR23021-61] - BSA and Azide free (ab272394)

View more associated products

Overview

  • Product name

    Anti-Ly-6K antibody [EPR23021-61]
    See all Ly-6K primary antibodies
  • Description

    Rabbit monoclonal [EPR23021-61] to Ly-6K
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, IPmore details
    Unsuitable for: Flow Cyt or ICC/IF
  • Species reactivity

    Reacts with: Human
  • Immunogen

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

  • Positive control

    • WB: HeLa cell lysate; Human testis tissue lysate. IHC-P:Human testis tissue. IP: Human testis tissue lysate.
  • 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: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR23021-61
  • Isotype

    IgG
  • Research areas

    • Immunology
    • Adaptive Immunity
    • B Cells
    • Non-CD
    • Immunology
    • Adaptive Immunity
    • T Cells
    • Non-CD

Associated products

  • Alternative Versions

    • Anti-Ly-6K antibody [EPR23021-61] - BSA and Azide free (ab272394)
  • Compatible Secondaries

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human LY6K (Ly-6K) knockout HeLa cell line (ab265874)
  • KO cell lysates

    • Human LY6K (Ly-6K) knockout HeLa cell lysate (ab263248)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab246486 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB
1/1000. Detects a band of approximately 24 kDa (predicted molecular weight: 19 kDa).
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP
1/30.
Notes
WB
1/1000. Detects a band of approximately 24 kDa (predicted molecular weight: 19 kDa).
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP
1/30.
Application notes
Is unsuitable for Flow Cyt or ICC/IF.

Target

  • Relevance

    The Lymphocyte Antigen 6 Complex is a 15 kDa phosphatidyl inositol anchored molecule, attached to the cell membrane by a GPI-anchor. The Lymphocyte Antigen 6 Complex is present on most immature T cells of the thymus, on lymphoid cells in bone marrow and peripheral B cells, but absent on peripheral T cells.
  • Cellular localization

    Cell Membrane
  • Database links

    • Entrez Gene: 54742 Human
    • Omim: 615093 Human
    • SwissProt: Q17RY6 Human
    • Alternative names

      • Lymphocyte antigen Ly-6K antibody
      • 2410015A16Rik antibody
      • 3110035B01Rik antibody
      • CT97 antibody
      • FLJ35226 antibody
      • HSJ001348 antibody
      • ly 6K antibody
      • Lymphocyte Antigen 6 Complex antibody
      • Lymphocyte antigen 6 complex, locus K antibody
      • mLy 6K antibody
      • ThB antibody
      • Thymocyte B cell antigen antibody
      see all

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ly-6K antibody [EPR23021-61] (ab246486)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ly-6K antibody [EPR23021-61] (ab246486)

      Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling Ly-6K with ab246486 at 1/2000 dilution followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in spermatocytes of human testis (PMID:18089789). The section was incubated with ab246486 for 30 mins at room temperature.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 Goat Anti-Rabbit IgG H&L (HRP).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

    • Western blot - Anti-Ly-6K antibody [EPR23021-61] (ab246486)
      Western blot - Anti-Ly-6K antibody [EPR23021-61] (ab246486)
      All lanes : Anti-Ly-6K antibody [EPR23021-61] (ab246486) at 1/1000 dilution

      Lane 1 : Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
      Lane 2 : LY6K knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

      Predicted band size: 19 kDa
      Observed band size: 24 kDa why is the actual band size different from the predicted?



      Lanes 1-2: Merged signal (red and green). Green - ab246486 observed at 24 kDa. Red - loading control ab8245 observed at 36 kDa.

       ab246486 Anti-Ly-6K antibody [EPR23021-61]  was shown to specifically react with Ly-6K in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265874 (knockout cell lysate ab263248) was used. Wild-type and Ly-6K knockout samples were subjected to SDS-PAGE. ab246486 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-Ly-6K antibody [EPR23021-61] (ab246486)
      Western blot - Anti-Ly-6K antibody [EPR23021-61] (ab246486)
      All lanes : Anti-Ly-6K antibody [EPR23021-61] (ab246486) at 1/1000 dilution

      Lane 1 : Human testis tissue lysate
      Lane 2 : Human heart tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : VeriBlot for IP secondary antibody(HRP)(ab131366) at 1/1000 dilution

      Predicted band size: 19 kDa
      Observed band size: 24 kDa why is the actual band size different from the predicted?



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

      The expression profile observed is consistent with what has been described in the literature (PMID: 27197181, 18089789).

      Negative control: Human heart (PMID:18089789)  Exposure time: 10 seconds

    • Immunoprecipitation - Anti-Ly-6K antibody [EPR23021-61] (ab246486)
      Immunoprecipitation - Anti-Ly-6K antibody [EPR23021-61] (ab246486)

      Ly-6K was immunoprecipitated from 0.35 mg Human testis tissue lysate with ab246486 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab246486 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/1000 dilution.

      Lane 1: Human testis tissue lysate 10 ug

      Lane 2: ab246486 IP in Human testis tissue lysate

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab246486 in human testis tissue lysate

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

      Exposure time: 3 seconds.

       

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ly-6K antibody [EPR23021-61] (ab246486)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ly-6K antibody [EPR23021-61] (ab246486)

      Immunohistochemical analysis of paraffin-embedded Human heart tissue labeling Ly-6K with ab246486 at 1/2000 dilution followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Negative control: No staining in human heart (PMID:18089789). The section was incubated with ab246486 for 30 mins at room temperature.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 Goat Anti-Rabbit IgG H&L (HRP).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

    • Anti-Ly-6K antibody [EPR23021-61] (ab246486)
      Anti-Ly-6K antibody [EPR23021-61] (ab246486)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    References (0)

    Publishing research using ab246486? Please let us know so that we can cite the reference in this datasheet.

    ab246486 has not yet been referenced specifically in any publications.

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