- Datasheet
- Specific References
- Protocols
Overview
- Product nameAnti-MED30 / TRAP25 antibody
See all MED30 / TRAP25 primary antibodies - DescriptionRabbit polyclonal to MED30 / TRAP25
- Tested applicationsSuitable for: ICC/IF, WBmore details
- Species reactivityReacts 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
- FormLiquid
- Storage instructionsShipped 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 bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 -
Concentration information loading... - PurityImmunogen affinity purified
- ClonalityPolyclonal
- IsotypeIgG
- Research areas
Associated products
- Compatible Secondaries
- Immunizing Peptide (Blocking)
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
- FunctionComponent 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 specificityExpressed in brain, heart, kidney, liver, lung, pancreas, placenta and skeletal muscle.
- Sequence similaritiesBelongs to the Mediator complex subunit 30 family.
- Cellular localizationNucleus.
- Information by UniProt
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Database links
- Entrez Gene: 613879 Cow
- Entrez Gene: 90390 Human
- Omim: 610237 Human
- SwissProt: Q2YDF2 Cow
- SwissProt: Q96HR3 Human
- Unigene: 492612 Human
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Alternative names
- MED30 antibody
- MED30_HUMAN antibody
- Mediator complex subunit 30 antibody
see all
Anti-MED30 / TRAP25 antibody images
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Immunocytochemistry/ Immunofluorescence - Anti-MED30 / TRAP25 antibody (ab49456)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.
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Western blot - Anti-MED30 / TRAP25 antibody (ab49456)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 minutesThis 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.
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Western blot - MED30 / TRAP25 antibody (ab49456)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 for Anti-MED30 / TRAP25 antibody (ab49456)
ab49456 has not yet been referenced specifically in any publications.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"