Overview

  • Product name

    Anti-PI4 kinase beta/PI4KB antibody
    See all PI4 kinase beta/PI4KB primary antibodies
  • Description

    Rabbit polyclonal to PI4 kinase beta/PI4KB
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IPmore details
    Unsuitable for: WB
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat, Rabbit, Horse, Chicken, Guinea pig, Cow, Dog, Turkey, Chimpanzee, Zebrafish, Rhesus monkey, Gorilla, Orangutan, Xenopus tropicalis
  • Immunogen

    Synthetic peptide corresponding to Human PI4 kinase beta/PI4KB aa 500-550. (CAA09495.1)

  • Positive control

    • HeLa whole cell lysates
  • General notes

     This product was previously labelled as PI4 kinase beta

     

Properties

Applications

Our Abpromise guarantee covers the use of ab135157 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
IP Use 2-10 µg for 1000 µg of chromatin.
  • Application notes
    Is unsuitable for WB.
  • Target

    • Function

      Phosphorylates phosphatidylinositol (PI) in the first committed step in the production of the second messenger inositol-1,4,5,-trisphosphate (PIP). May regulate Golgi disintegration/reorganization during mitosis, possibly via its phosphorylation. Involved in Golgi-to-plasma membrane trafficking.
    • Tissue specificity

      Widely expressed with highest levels in heart, skeletal muscle, pancreas, testis and ovary. Weakly expressed in liver.
    • Sequence similarities

      Belongs to the PI3/PI4-kinase family. Type III PI4K subfamily.
      Contains 1 PI3K/PI4K domain.
      Contains 1 PIK helical domain.
    • Cellular localization

      Endomembrane system. Mitochondrion outer membrane. Rough endoplasmic reticulum membrane. Golgi apparatus. Cytoplasm, perinuclear region. Found in the outer membrane of mitochondria and membranes of the rough endoplasmic reticulum. Recruited to the Golgi complex by the small GTPase ARF to stimulate the synthesis of phosphatidylinositol 4,5-bisphosphate (PIP2) on the Golgi complex.
    • Information by UniProt
    • Database links

    • Alternative names

      • DKFZp686O1820 antibody
      • FLJ30129 antibody
      • NPIK antibody
      • Phosphatidylinositol 4 kinase beta antibody
      • Phosphatidylinositol 4 kinase catalytic beta antibody
      • Phosphatidylinositol 4 kinase catalytic beta polypeptide antibody
      • Phosphatidylinositol 4 kinase wortmannin sensitive antibody
      • Phosphatidylinositol 4-kinase beta antibody
      • PI4K BETA antibody
      • PI4K-beta antibody
      • PI4K92 antibody
      • PI4KB antibody
      • PI4KB_HUMAN antibody
      • PI4Kbeta antibody
      • PI4KIIIbeta antibody
      • PIK4CB antibody
      • PtdIns 4 kinase antibody
      • PtdIns 4-kinase beta antibody
      • Type III phosphatidylinositol 4 kinase beta antibody
      see all

    Images

    • Lane 1: Immunoprecipitation from HeLa whole cell lysate (1 mg for IP, 20% of IP loaded), using an antibody which recognized an upstream epitope of PI4 kinase beta/PI4KB.
      Lane 2: Detection of Human PI4 kinase beta/PI4KB by Immunoprecipitation from HeLa whole cell lysate (1 mg for IP, 20% of IP loaded), using ab135157 at 6 µg/mg lysate for IP.
      Lane 3: Immunoprecipitation from HeLa whole cell lysate (1 mg for IP, 20% of IP loaded), using Control IgG.
      Subsequent Western blot detection using an antibody which recognized an upstream epitope of PI4 kinase beta/PI4KB.
      Developed using the ECL technique with an exposure time of 3 seconds.

    References

    ab135157 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

    Question
    Answer

    The laboratory have provided the protocol which is copied below. Please note that this would be a guideline only and may require individual optimization.


    Protocol:

    Reagents Needed:

    Immunoprecipitation (IP) lysis buffer
    Protease Inhibitors
    Primary Antibodies made in Rabbit
    Normal IgG, negative control (Rabbit IgG isotype control)
    Protein A Sepharose Beads
    Cell Lysate
    Sample Buffer

    IP lysis Buffer
    12.5 ml 1M NaCl (250mM)
    2.5 ml 1M Tris (50mM)
    500ul 0.5M EDTA (5mM)
    2.5ml 10% NP-40
    32 ml dH20

    Protein A Beads
    Resuspend 400 mg of Protein A beads in 10 ml of distilled H2O. Mix well to resuspend. Spin at 250 rpm for 5 minutes. Wash 3X in 10 ml IP Lysis buffer. Resuspend to 10 ml with IP lysis buffer for a 20% solution. Use 100 mcl per IP reaction.

    4X Sample Buffer
    Glycerol 4.0 g
    Tris Base 0.68 g
    Tris HCL 0.67 g
    LDS 0.80 g
    EDTA 6 mg
    Brilliant Blue G250 2.5 mg
    Phenol red 2.5 mg
    Adjust volume to 10 ml with ultra pure water.

    Store at 4 C.

    1X Sample Buffer
    4X sample buffer 150 mcl
    1M DTT 60 mcl
    Distilled water 390 mcl

    Make fresh for each use.

    Procedure:

    Prepare cell lysate according to protocol. Place 500 mcl of the prepared cell lysate (1-3 mg/ml) into a 1.5 ml micro-centrifuge tube.
    To this tube add 2 to 10 mcg of the primary antibody (If using neat sera or an IgG fraction such as Protein-A purified antibody, larger amounts are likely to be required. For best results, optimal amounts of antibody should be empirically defined.)
    To a negative control reaction, add an equivalent amount of normal rabbit IgG.
    Add 100 mcl of a 20% Protein A suspension to the mixture of antibody and cell lysate. Rotate the immunoprecipitation reactions (end-to-end) for 3 hours at room temperature or overnight at 4 C.
    Centrifuge (200 x g; 5 minutes) to pellet the complex.
    Remove the supernatant and add 500 mcl cold cell lysis buffer. Centrifuge (200 x g; 5 minutes).
    Repeat wash step 6 twice more. After each centrifugation remove as much of the supernatant as possible.
    After removing the supernatant from the third wash, add 40 mcl of freshly prepared 1X sample buffer to each tube and heat at 90 C for 5 minutes.
    Continue with electrophoresis and immunoblotting as described under western blotting protocol. Load 8 to 16 mcl (20 to 40% of the IP reaction) to a polyacrylamide gel.



    Note: For optimal results, complete reduction of the sample is required. We recommend the use of 0.1 M DTT in SDS-PAGE sample buffer and immediately heating samples, loading and running gels.

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