Human DEPTOR/DEPDC6 Antibody

Catalog #: AF7255 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF7255
AF7255-SP

Detection of Human DEPTOR/DEPDC6 by Western Blot.

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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human DEPTOR/DEPDC6 Antibody Summary

Species Reactivity

Human

Specificity

Detects recombinant human DEPTOR/DEPDC6 in direct ELISAs and Western blots.

Source

Polyclonal Sheep IgG

Purification

Antigen Affinity-purified

Immunogen

E. coli-derived recombinant human DEPTOR/DEPDC6
Ala31-Gly158
Accession # Q8TB45

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 DEPTOR/DEPDC6 by Western Blot. Western blot shows lysates of HT-29 human colon adenocarcinoma cell line and IMR-32 human neuroblastoma cell line. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human DEPTOR/DEPDC6 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7255) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for DEPTOR/DEPDC6 at approximately 50 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.

Immunohistochemistry

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DEPTOR/DEPDC6 in Human Kidney. DEPTOR/DEPDC6 was detected in immersion fixed paraffin-embedded sections of human kidney using Sheep Anti-Human DEPTOR/DEPDC6 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7255) 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-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific staining was localized to epithelial cells in convoluted tubules. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Sterile PBS to a final concentration of 0.2 mg/mL.

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: DEPTOR/DEPDC6

DEPDC6 (DEP domain-containing protein 6; also DEPTOR) is a 47-48 kDa DEP (Disheveled/Egl10/Pleckstrin) domain-containing protein found in a wide variety of cells. It is a component of the IGF/Ins activated mTOR complexes 1 and 2 (TORC1 and 2) that promote S6K1 and Akt phosphorylation, respectively. Within TORC1, DEPDC6 acts as a natural inhibitor of mTOR kinase, suppressing S6K1 activity. Conversely, this inhibition of mTOR promotes TORC2 activation and Akt activity. In general, a decline in DEPDC6 activity promotes both an increase in cell size and lifespan. Human DEPDC6 is 409 amino acids (aa) in length. It is acetylated on Met1, contains two DEP domains (aa 36-119 and 145-219), and ends with a mTOR-interacting PDZ domain (aa 330-407). There are at least 13 utilized Ser/Thr phosphorylation sites. Although no splice variants have been reported, the highly orthologous mouse form of DEPDC6 does show a 34 aa substitution for the C-terminal 42 aa, suggesting that this splicing event may occur in human. Over aa 31-158, human DEPDC6 shares 98% aa sequence identity with mouse DEPDC6.

Long Name

DEP Domain Containing mTOR-interacting Protein

Entrez Gene IDs

64798 (Human); 97998 (Mouse)

Alternate Names

DEP domain containing 6; DEP domain containing mTOR interacting protein; DEP domain containing MTOR-interacting protein; DEP domain-containing mTOR-interacting protein; DEP domain-containing protein 6; DEP.6; DEPDC6; DEPDC6FLJ12341; DEPTOR; DKFZp564B1778; FLJ12428; FLJ13854