Overview

  • Product name
    Anti-MED30 / TRAP25 antibody
    See all MED30 / TRAP25 primary antibodies
  • Description
    Rabbit polyclonal to MED30 / TRAP25
  • Tested applications
    Suitable for: ICC/IF, WBmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Cow, Dog, Chimpanzee
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human MED30/ TRAP25.

    (Peptide available as ab71170.)

  • Positive control
    • This antibody gave a positive signal in the following Human Lysates: HCT 116 Whole Cell HEK293 Whole Cell TE 671 Whole Cell This antibody gave a positive result when used in the following methanol fixed cell lines: HepG2

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 or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    Preservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab49456 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 5 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 22 kDa (predicted molecular weight: 20 kDa).

Target

  • Function
    Component of the Mediator complex, a coactivator involved in the regulated transcription of nearly all RNA polymerase II-dependent genes. Mediator functions as a bridge to convey information from gene-specific regulatory proteins to the basal RNA polymerase II transcription machinery. Mediator is recruited to promoters by direct interactions with regulatory proteins and serves as a scaffold for the assembly of a functional preinitiation complex with RNA polymerase II and the general transcription factors.
  • Tissue specificity
    Expressed in brain, heart, kidney, liver, lung, pancreas, placenta and skeletal muscle.
  • Sequence similarities
    Belongs to the Mediator complex subunit 30 family.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • MED30 antibody
    • MED30_HUMAN antibody
    • Mediator complex subunit 30 antibody
    • Mediator of RNA polymerase II transcription subunit 30 antibody
    • THRAP6 antibody
    • Thyroid hormone receptor-associated protein 6 antibody
    • Thyroid hormone receptor-associated protein complex 25 kDa component antibody
    • TRAP/Mediator complex component TRAP25 antibody
    • Trap25 antibody
    see all

Images

  • ICC/IF image of ab49456 stained HepG2 cells. The cells were 100% methanol fixed (5 min) then permeabilised using 0.1% PBS-Triton and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to further permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab49456 at 1µg/ml overnight at +4°C. The secondary antibody (pseudo-colored green) was Alexa Fluor® 488 goat anti- rabbit (ab150081) IgG (H+L) preadsorbed, used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1h at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.

  • All lanes : Anti-MED30 / TRAP25 antibody (ab49456) at 1 µg/ml

    Lane 1 : HCT 116 (Human Colorectal Carcinoma) Whole Cell Lysate
    Lane 2 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 3 : TE 671 (Human Rhabdomyosarcoma) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/50000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 20 kDa
    Observed band size : 22 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 19 kDa,85 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 8 minutes

    This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab49456 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

  • All lanes : Anti-MED30 / TRAP25 antibody (ab49456) at 1 µg/ml

    Lane 1 : HCT 116 (Human Colorectal Carcinoma) Whole Cell Lysate
    Lane 2 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 3 : TE 671 (Human Rhabdomyosarcoma) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 20 kDa
    Observed band size : 19 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 250 kDa,75 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 18 minutes

References

ab49456 has not yet been referenced specifically in any publications.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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