Human MED4 Antibody

Catalog #: AF5089 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF5089
AF5089-SP

Detection of Human MED4 by Western Blot.

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Human MED4 Antibody Summary

Species Reactivity

Human

Specificity

Detects endogenous human MED4 in Western blots.

Source

Polyclonal Goat IgG

Purification

Antigen Affinity-purified

Immunogen

E. coli-derived recombinant human MED4
Met80-Asp261
Accession # Q9NPJ6

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Applications

Recommended Concentration

Sample

Western Blot

1 µg/mL

See below

Immunohistochemistry

5-15 µg/mL

See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot

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Detection of Human MED4 by Western Blot. Western blot shows lysates of Jurkat human acute T cell leukemia cell line, K562 human chronic myelogenous leukemia cell line, and JEG-3 human epithelial choriocarcinoma cell line. PVDF membrane was probed with 1 µg/mL of Human MED4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5089) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for MED4 at approximately 41 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 3.

Immunohistochemistry

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MED4 in Human Prostate. MED4 was detected in immersion fixed paraffin-embedded sections of human prostate using Goat Anti-Human MED4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5089) at 3 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to nuclei in glandular epithelial cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: MED4

MED4 (Mediator complex 4 subunit; also DRIP36) is a 34‑36 kDa member of the mediator complex subunit 4 family of proteins. It is part of a 20+ subunit complex named Mediator that serves as a bridge between RNA polymerase II and DNA binding regulatory proteins that cooperate during RNA synthesis. Human MED4 is 270 amino acids (aa) in length and contains two N-terminal coiled-coil regions (aa 24‑48 and 90‑131) plus a C-terminal poly-Ser region (aa 262‑269). Multiple splice variants may exist. There is an alternate start site at Met47 that may be accompanied by a premature truncation after Asp239. There is also a form with a six aa substitution for aa 105‑270, and a fourth form that shows a 19 aa substitution for aa 1‑41, accompanied by a truncation after Thr260. Over aa 80‑261, human MED4 is 97% aa identical to mouse MED4.

Long Name

Mediator Complex Subunit 4

Entrez Gene IDs

29079 (Human); 67381 (Mouse); 306030 (Rat)

Alternate Names

Activator-recruited cofactor 36 kDa component; ARC36; DRIP36; DRIP36mediator of RNA polymerase II transcription, subunit 4 homolog (S. cerevisiae); FLJ10956; HSPC126; HSPC126vitamin D receptor interacting protein; MED4; mediator complex subunit 4TRAP/SMCC/PC2 subunit p36 subunit; subunit 4 homolog; TRAP36; VDRIP