Human/Mouse/Rat PDK‑1 Antibody

Name: Human/Mouse/Rat PDK‑1 Antibody
Brand: R&D Systems
SKU: MAB864
Price: 449 USD
Availability: InStock

R&D Systems | Catalog # MAB864

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Citations (4)

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Product Specifications
Scientific Data
Applications
Preparation & Storage
Calculators
Background
Product Documents
Specific Notices
Research Areas

Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Mouse

Applications

Validated:

Western Blot, Immunocytochemistry, Simple Western

Cited:

Immunohistochemistry, Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG_2A Clone # 650308

View all PDK-1 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human PDK-1
Asn411-Gln556
Accession # O15530

Specificity

Detects human, mouse, and rat PDK-1 in Western blots.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG_2A

Scientific Data Images for Human/Mouse/Rat PDK‑1 Antibody

Detection of Human, Mouse, and Rat PDK‑1 by Western Blot.

Western blot shows lysates of Jurkat human acute T cell leukemia cell line, HepG2 human hepatocellular carcinoma cell line, Daudi human Burkitt's lymphoma cell line, A20 mouse B cell lymphoma cell line, and C6 rat glioma cell line. PVDF Membrane was probed with 1 µg/mL of Mouse Anti-Human/Mouse/Rat PDK-1 Monoclonal Antibody (Catalog # MAB864) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF007). Specific bands were detected for PDK-1 at approximately 58-68 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

PDK‑1 in HeLa Human Cell Line.

PDK-1 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Mouse Anti-Human/Mouse/Rat PDK-1 Monoclonal Antibody (Catalog # MAB864) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (yellow; NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of Human PDK‑1 by Simple Western^TM.

Simple Western shows lysates of Daudi human Burkitt's lymphoma cell line and HepG2 human hepatocellular carcinoma cell line, loaded at 0.5 mg/ml. A specific band was detected for PDK‑1 at approximately 60 kDa (as indicated) using 20 µg/mL of Mouse Anti-Human/Mouse/Rat PDK‑1 Monoclonal Antibody (Catalog # MAB864). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Applications for Human/Mouse/Rat PDK‑1 Antibody

Application

Recommended Usage

Immunocytochemistry

5-25 µg/mL
Sample: Immersion fixed HeLa human cervical epithelial carcinoma cell line

Simple Western

20 µg/mL
Sample: Daudi human Burkitt's lymphoma cell line and HepG2 human hepatocellular carcinoma cell line

Western Blot

1 µg/mL
Sample: Jurkat human acute T cell leukemia cell line, HepG2 human hepatocellular carcinoma cell line, Daudi human Burkitt's lymphoma cell line, A20 mouse B cell lymphoma cell line, and C6 rat glioma cell line

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Sterile PBS to a final concentration of 0.5 mg/mL. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.
*Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in 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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

Volume (to add to vial)

Mass (in vial)

Desired Reconstitution Concentration

Background: PDK-1

PDK-1 (3-phosphoinositide-dependent protein kinase, gene name PDPK1) is a 58 -68 kDa, 556 amino acid (aa) monomeric protein of the AGC serine/threonine kinase family. It is activated by phosphorylation in the presence of PtdIns(3,4,5) P3 or PtdIns(3,4) P2. Akt, S6 kinases, PKA and PKC-zeta are reported PDK-1 substrates. Through Akt, PDK-1 mediates many of the intracellular actions of insulin. Within the region used as an immunogen, human PDK-1 shares 98% aa identity with mouse and rat PDK-1. One reported isoform has an alternate start site at aa 50, while another lacking aa 238-263 is predicted to be catalytically inactive.