Overview

  • Product name

  • Description

    Rabbit polyclonal to KDS
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IPmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat, Rabbit, Horse, Guinea pig, Cow, Dog, Pig, Xenopus laevis, Rhesus monkey, Gorilla, Chinese hamster, Orangutan, Elephant
  • Immunogen

    Synthetic peptide corresponding to Human KDS (C terminal).
    Database link: Q9H2K8

  • Positive control

    • HeLa whole cell lysates.
  • General notes

     This product was previously labelled as TAOK3

     

Properties

Applications

Our Abpromise guarantee covers the use of ab70297 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
IP
  • Application notes
    IP: Use at 1-5µg/mg of lysate.
    WB: 1/5000 - 1/25000. Detects a band of approximately 120 kDa (predicted molecular weight: 105 kDa).


    Not yet tested in other applications.
    Optimal dilutions/concentrations should be determined by the end user.
  • Target

    • Function

      Inhibits the basal activity of Jun kinase. Negatively regulated by epidermal growth factor (EGF). When overexpressed, may activate ERK1/ERK2 and JNK/SAPK.
    • Tissue specificity

      Ubiquitously expressed at a low level, and highly expressed in peripheral blood leukocytes (PBLs), thymus, spleen, kidney, skeletal muscle, heart and liver.
    • Sequence similarities

      Belongs to the protein kinase superfamily. STE Ser/Thr protein kinase family. STE20 subfamily.
      Contains 1 protein kinase domain.
    • Post-translational
      modifications

      Autophosphorylated. Phosphorylated upon DNA damage, probably by ATM or ATR.
    • Cellular localization

      Cytoplasm. Cell membrane. Also localized to the peripheral cell membrane.
    • Information by UniProt
    • Database links

    • Alternative names

      • CTCL tumor antigen HD CL 09 antibody
      • CTCL-associated antigen HD-CL-09 antibody
      • Cutaneous T cell lymphoma tumor antigen HD CL 09 antibody
      • Cutaneous T-cell lymphoma-associated antigen HD-CL-09 antibody
      • Dendritic cell derived protein kinase antibody
      • Dendritic cell-derived protein kinase antibody
      • DKFZp666H245 antibody
      • DPK antibody
      • FLJ31808 antibody
      • hKFC A antibody
      • hKFC-A antibody
      • JIK antibody
      • JNK/SAPK inhibitory kinase antibody
      • JNK/SAPK-inhibitory kinase antibody
      • Jun kinase inhibitory kinase antibody
      • Jun kinase-inhibitory kinase antibody
      • KDS antibody
      • Kinase from chicken homolog A antibody
      • MAP3K18 antibody
      • Serine kinase antibody
      • Serine/threonine-protein kinase TAO3 antibody
      • STE20 like kinase antibody
      • TAO kinase 3 antibody
      • TAOK3 antibody
      • TAOK3_HUMAN antibody
      • Thousand and one amino acid protein 3 antibody
      see all

    Images

    • All lanes : Anti-KDS antibody (ab70297) at 0.1 µg/ml

      Lane 1 : HeLa whole cell lysate at 50 µg
      Lane 2 : HeLa whole cell lysate at 15 µg

      Developed using the ECL technique.

      Predicted band size: 105 kDa
      Observed band size: 120 kDa
      why is the actual band size different from the predicted?
      Additional bands at: 45 kDa. We are unsure as to the identity of these extra bands.


      Exposure time: 3 minutes
    • Western Blot showing Lane 2: ab70297 staining JIK in HeLa whole cell lysates immunoprecipitated using ab70297 at 3µg/mg lysate (1mg/lane). Lane 1: control IgG used for immunoprecipitation.Detection: Chemiluminescence with an exposure time of 3 minutes

    References

    ab70297 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

    1-4 of 4 Abreviews or Q&A

    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (Murine embryonic fibroblast)
    Gel Running Conditions
    Non-reduced Denaturing (gel 10%)
    Loading amount
    20 µg
    Treatment
    siRNA trasnfection directed against TAOK3
    Specification
    Murine embryonic fibroblast
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

    Bastiaan Maes

    Verified customer

    Submitted May 02 2016

    Question

    Dear,

    Hereby I sent you the completed questionnaire about the TAOK3 Ab.


    Order Details Antibody code: ab 70297 Problem Non-specific band Multiple bands No signal or weak signal High background Lot number lot GR44200-2 Purchase order number or preferably Abcam order number:

    General Information Antibody storage conditions (temperature/reconstitution etc)
    -20oC, aliquoted in small batches
    Description of the problem (high background, wrong band size, more bands, no band etc.)
    high background, wrong band size, no band on expected MW size
    Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.) cell extracts from different cell types (organs, MEF…)
    Sample preparation (Buffer/Protease inhibitors/Heating sample etc.)
    E1A buffer (1% NP40) + protease inhibitors + sample buffer
    5min at 95°C Amount of protein loaded ± 20-30µg Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.) 10% SDS gel Transfer and blocking conditions (Buffer/time period, Blocking agent etc.) wet blot transfer 1-2hrs 4°C Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step) Rb polyclonal Ab to JIK
    1/3000 in 3% BSA in TBS-tween overnight
    3 washes 15min
    Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step) goat anti rabbit (H+L chain HRP conjugated from Jackson Laboratories
    1/30000 in 5% non fat milk in TBS-tween
    1hr room temperature
    3 wahes 15 min Detection method (ECL, ECLPlus etc.) ECL plus from Roche Positive and negative controls used (please specify) JIK KO samples as negative control
    JIK overexpressing cells as positive control Optimization attempts (problem solving) How many times have you tried the Western? 3 times Have you run a "No Primary" control? Yes No Do you obtain the same results every time? Yes No e.g. are the background bands always in the same place? What steps have you altered? we tried 3% BSA instead off 5% non fat milk Additional Notes: Image: We would appreciate if you are able to provide an image (including molecular weight markers) which would help us to asess the results.

    Read More
    Answer

    Thank you for taking the time to complete our questionnaire as requested.

    The details you have kindly provided will enable us to investigate this case for you and this is also helpful in our records for monitoring of quality.

    Reviewing this case, I would like to offer some suggestions to help optimize the results. I would also appreciate if you can confirm some further details:

    1. Could you confirm if the wash steps? I can suggest to wash in TBS containing 0.2% Tween if this has not already been tried. TBS can provide a more stringent wash than PBS and the Tween will help to wash away any excess antibody.

    2. We recommend not to mix blocking agents in one experiment. Try BSA only.

    3. Is the secondary antibody and detection kit working well with other primary antibodies? What are the results from the no primary control? This would help to assess if the secondary antibody is binding non specifically. The concentration of secondary antibody may need to be reduced in order to help optimize the results.

    4. I am sorry to letyou know that the attachment with the image has not come through. I would appreciate if you are able to send this again.

    5, I have found several slightly different recipes for EIA buffer. Could you confirm which recipe you have used? I can suggest to consider trying RIPA lysis buffer instead which may provide a more suitable protein preparation in this case.

    I hope this information is helpful, thank you for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details.

    Read More

    Answer

    Thank you for taking the time to contact us and for your telephone call yesterday afternoon. I appreciate your sending a message, I did in factunfortunately have your email address incorrectand am pleased to have confirmation of this.

    I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody. I would like to reassure you that this antibody is tested and covered by our 6 month guarantee forWB and human. In the event that a product is not functioning in the tested applications and species cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund.

    As discussed on the phone,we would like to investigate thiscase further for you, and also obtain more information for our quality monitoring records. In order to proceed with this, I have enclosed a technical questionnaire below. I would appreciate if you could complete this. It will help you put the information we require together very easily.

    Thank you for providing an image alreadywhich would help us to assess the results. I will be away from the office next week, but one of my colleagues will be able to investigate this for you and get back to you.

    Thank you for your time and cooperation. We look forward to receiving the completed questionnaire.


    Order Details
    Antibody code:

    Problem
    Choose: Non-specific band Multiple bands No signal or weak signal High background

    Lot number

    Purchase order number
    or preferably Abcam order number:



    General Information
    Antibody storage conditions (temperature/reconstitution etc)


    Description of the problem (high background, wrong band size, more bands, no band etc.)


    Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.)


    Sample preparation (Buffer/Protease inhibitors/Heating sample etc.)


    Amount of protein loaded


    Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.)


    Transfer and blocking conditions (Buffer/time period, Blocking agent etc.)


    Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


    Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


    Detection method (ECL, ECLPlus etc.)


    Positive and negative controls used (please specify)



    Optimization attempts (problem solving)
    How many times have you tried the Western?



    Have you run a "No Primary" control?
    Yes No

    Do you obtain the same results every time?
    Yes No
    e.g. are the background bands always in the same place?


    What steps have you altered?


    Additional Notes:


    Image:
    We would appreciate if you are able to provide an image (including molecular weight markers) which would help us to asess the results.

    Read More
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (Spleen)
    Gel Running Conditions
    Reduced Denaturing (6% running section)
    Loading amount
    100 µg
    Specification
    Spleen
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

    Abcam user community

    Verified customer

    Submitted May 18 2009

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