Anti-IKK alpha (phospho T23) antibody (ab38515)

Reviews (2)Q&A (3)References (18)

Overview

  • Product name

    Anti-IKK alpha (phospho T23) antibody
    See all IKK alpha primary antibodies

  • Description

    Rabbit polyclonal to IKK alpha (phospho T23)

  • Host species

    Rabbit

  • Specificity

    This antibody is specific for IKK alpha only when phosphorylated at threonine 23.

  • Tested applications

    Suitable for: ICC/IF, WB, IHC-P, ELISA, IHC-Frmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic phosphopeptide derived from Human IKK alpha around the phosphorylation site of threonine 23 (LGTpGG).

  • Positive control

    • Colon carcinoma tissue slides and MDA-MB-435 cell extract.

Properties

Applications

Our Abpromise guarantee covers the use of ab38515 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
ICC/IF Use a concentration of 1 µg/ml.
WB 1/500 - 1/1000. Detects a band of approximately 85 kDa (predicted molecular weight: 85 kDa).
IHC-P Use at an assay dependent concentration.
ELISA 1/4000.
IHC-Fr Use at an assay dependent concentration. See Abreview.

Target

  • Function

    Acts as part of the IKK complex in the conventional pathway of NF-kappa-B activation and phosphorylates inhibitors of NF-kappa-B thus leading to the dissociation of the inhibitor/NF-kappa-B complex and ultimately the degradation of the inhibitor. As part of the non-canonical pathway of NF-kappa-B activation, the MAP3K14-activated CHUK/IKKA homodimer phosphorylates NFKB2/p100 associated with RelB, inducing its proteolytic processing to NFKB2/p52 and the formation of NF-kappa-B RelB-p52 complexes. Also phosphorylates NCOA3. Phosphorylates 'Ser-10' of histone H3 at NF-kappa-B-regulated promoters during inflammatory responses triggered by cytokines.

  • Tissue specificity

    Widely expressed.

  • Involvement in disease

    Defects in CHUK are the cause of cocoon syndrome (COCOS) [MIM:613630]; also known as fetal encasement syndrome. COCOS is a lethal syndrome characterized by multiple fetal malformations including defective face and seemingly absent limbs, which are bound to the trunk and encased under the skin.

  • Sequence similarities

    Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. I-kappa-B kinase subfamily.
    Contains 1 protein kinase domain.

  • Post-translational
    modifications

    Phosphorylated by MAP3K14/NIK, AKT and to a lesser extent by MEKK1, and dephosphorylated by PP2A. Autophosphorylated.
    Acetylation of Thr-179 by Yersinia yopJ prevents phosphorylation and activation, thus blocking the I-kappa-B signaling pathway.

  • Cellular localization

    Cytoplasm. Nucleus. Shuttles between the cytoplasm and the nucleus.
  • Information by UniProt

  • Database links

    • Alternative names

      • chuk antibody
      • CHUK1 antibody
      • Conserved Helix Loop Helix Ubiquitous Kinase antibody
      • Conserved helix loop ubiquitous kinase antibody
      • Conserved helix-loop-helix ubiquitous kinase antibody
      • I Kappa B Kinase 1 antibody
      • I Kappa B Kinase Alpha antibody
      • I-kappa-B kinase 1 antibody
      • I-kappa-B kinase alpha antibody
      • IkappaB kinase antibody
      • IkB kinase alpha subunit antibody
      • IkBKA antibody
      • IKK 1 antibody
      • IKK A antibody
      • IKK a kinase antibody
      • IKK-A antibody
      • IKK-alpha antibody
      • IKK1 antibody
      • IKKA antibody
      • IKKA_HUMAN antibody
      • Inhibitor Of Kappa Light Polypeptide Gene Enhancer In B Cells antibody
      • Inhibitor Of Nuclear Factor Kappa B Kinase Alpha Subunit antibody
      • Inhibitor of nuclear factor kappa-B kinase subunit alpha antibody
      • NFKBIKA antibody
      • Nuclear Factor Kappa B Inhibitor Kinase Alpha antibody
      • Nuclear factor NF kappa B inhibitor kinase alpha antibody
      • Nuclear factor NF-kappa-B inhibitor kinase alpha antibody
      • Nuclear factor NFkappaB inhibitor kinase alpha antibody
      • Nuclear Factor Of Kappa Light Chain Gene Enhancer In B Cells Inhibitor antibody
      • TCF-16 antibody
      • TCF16 antibody
      • Transcription factor 16 antibody
      see all

    Images

    • Western blot - Anti-IKK alpha (phospho T23) antibody (ab38515)
      Western blot - Anti-IKK alpha (phospho T23) antibody (ab38515)
      All lanes : Anti-IKK alpha (phospho T23) antibody (ab38515) at 1/500 dilution

      Lane 1 : MDA-MB-435 cell extract.
      Lane 2 : MDA-MB-435 cell extract + EGF ( The cells were treated with 200ng/ml EGF for 5 minutes).

      Lysates/proteins at 30 µg per lane.

      Secondary
      All lanes : Alkaline Phosphatase AffiniPure Goat Anti-Rabbit IgG (H+L)

      Predicted band size: 85 kDa
      Observed band size: 85 kDa



      Lanes can be loaded with 5-30µg of total protein.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IKK alpha (phospho T23) antibody (ab38515)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IKK alpha (phospho T23) antibody (ab38515)
      ab38515 at a 1:50 dilution staining IKK alpha in Human colon carcinoma tissue using Immunohistochemistry, Paraffin Embedded Tissue.

      Left image : untreated.

      Right image : treated with phosphopeptide.

    • Immunohistochemistry (Frozen sections) - Anti-IKK alpha (phospho T23) antibody (ab38515)
      Immunohistochemistry (Frozen sections) - Anti-IKK alpha (phospho T23) antibody (ab38515)This image is courtesy of an anonymous Abreview

       ab38515 at 1/100 staining mouse skin tissue sections by IHC-Fr. The tissue was paraformaldehyde fixed and blocked with serum prior to incubation with the antibody for 1 hour. An Alexa Fluor ® conjugated goat anti-rabbit antibody was used as the secondary.

      See Abreview

    • Immunocytochemistry/ Immunofluorescence - Anti-IKK alpha (phospho T23) antibody (ab38515)
      Immunocytochemistry/ Immunofluorescence - Anti-IKK alpha (phospho T23) antibody (ab38515)
      ICC/IF image of ab38515 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab38515, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    References

    This product has been referenced in:

    • Kim YH  et al. Expression of Nod-like Receptors and Clinical Correlations in Patients with Dry Eye Disease. Am J Ophthalmol N/A:N/A (2019). Read more (PubMed: 30653959) »
    • Jiang L  et al. Overexpression of PIMREG promotes breast cancer aggressiveness via constitutive activation of NF-?B signaling. EBioMedicine 43:188-200 (2019). Read more (PubMed: 30979686) »
    See all 18 Publications for this product

    Publishing research using ab38515? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    1-5 of 5 Abreviews or Q&A

    Question

    human skin tissue, IHC-P
    mouse skin tissue, IHC-Fr
    no signal at all
    Ab from other company gave desired staining
    same standard protocol for both Abs:
    AR: citrate buffer
    Ab: 1/25, 1/50, 1/75, 1/100 several incubation times
    secondary: HRP or polymer
    blocking: not sure

    Read More
    Answer

    Thank you for contacting us.

    Your credit note IDs are xxx and xxx.

    I am sorry that this antibody did not perform as stated on the datasheet. I have asked our accounting department to issue a credit note for you, which can be redeemed against the invoice of a future order by passing it on to your purchasing department. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used. If you have questions on how to use the credit note, please contact our accounting department.

    Our accounting department can be contacted by email at us.credits@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website. Please refer to the credit note ID in any correspondence with our accounting department.

    The credit note ID is for your reference only and does not automatically guarantee the credit.

    I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further expert advice.

    Read More

    Western blot abreview for Anti-IKK alpha (phospho T23) antibody

     Poor
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Guinea pig Tissue lysate - whole (myometrium pregnant and non-pregnant)
    Loading amount
    30 µg
    Specification
    myometrium pregnant and non-pregnant
    Gel Running Conditions
    Reduced Denaturing (10%)
    Blocking step
    Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 07 2012

    Question

    I used this antibody in WB of various human fibroblast cell lines. I detect a single band, but it is at 130kDa, rather than 85 - is this the correct band?

    Read More
    Answer

    Thank you for contacting us regarding ab38515.

    I have received the following additional information from the lab:

    This target containing leucine zipper associates with many proteins. It is posible that theband is dimerization. The leucine zipper is important to allow homo- and hetero-dimerization. Add DTT to lysis buffer as we find this helps.

    I hope this information is helpful. Please do not hesitate to contact us if you have any additional questions.

    Read More

    Question

    Dear Sir,
    I have a query regarding your IKK-alpha antibody (code-ab105923,product name-Anti-IKK alpha antibody [3G12H9])...I want to use this antibody for flow cytometric analysis of phosphorylated IKK-alpha, but do your antibody detect phosphorylated IKK-alpha or only IKK-alpha? becuase I want to study the TLR signalling pathway and for this I want to look at activated (phosphorylated) IIKK alpha enzyme. So can you guide me regarding this...so that I can order this antibody accordingly?

    Waiting for you reply..
    Thank you.

    Read More
    Answer

    Thank you for your enquiry.

    I can confirm that as ab105923IKK alpha antibodyhas not specifically been raised against a phosphorylated residue of the protein, it is therefore likely to detect both phosphorylated and non phosphorylated forms of the protein.

    We do have the following antibodies that detect some of the phosphorylated residues specifically. I can recommend to review the datasheets for further details and to select one tested in the species you wil be using.

    ab38515 Anti-IKK alpha (phospho T23) antibody
    Rabbit polyclonal
    Applications: ELISA, ICC/IF, IHC-Fr, IHC-P, WB
    Reacts with: Human, Mouse, Rat
    https://www.abcam.com/index.html?datasheet=38515 (or use the following: https://www.abcam.com/index.html?datasheet=38515).

    ab17943 Anti-IKK alpha (phospho S176 + S180) antibody
    Rabbit polyclonal
    Applications: ICC/IF, WB
    Reacts with: Human
    https://www.abcam.com/index.html?datasheet=17943 (or use the following: https://www.abcam.com/index.html?datasheet=17943).

    I am sorry that as far as we are aware, these have not been tested and therefore are not guaranteed in flow cytometry.However, if you would like to test these antibodies in flow cystometry, please do not hesitate to contact me again prior to the purchase by replying to this message as you may be eligible for our testing discount program.

    Otherwise, we like to encourage all of our customers to submit an Abreview via the online product datasheet. We always appreciate customer feedback, whether positive or negative, and we make all product information available to researchers. Plus, each Abreview earns Abpoints that can be used for discounts on future purchases or rewards such as Amazon.com gift certificates.

    To find out more about our Abreview system, please see the following link:

    https://www.abcam.com/abreviews

    I hope this information is helpful. Please do not hesitate to contact me for any further advice or information.

    Read More

    Immunohistochemistry (Frozen sections) abreview for Anti-IKK alpha (phospho T23) antibody

     Excellent
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (skin)
    Specification
    skin
    Fixative
    Paraformaldehyde
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 6%
    Read More

    Abcam user community

    Verified customer

    Submitted Apr 24 2007

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