Human phospho-p70 S6 Kinase (T229) Antibody (1015B)

Name: Human phospho-p70 S6 Kinase (T229) Antibody (1015B)
Brand: R&D Systems
SKU: MAB8964
Price: 469 USD
Availability: InStock

R&D Systems | Catalog # MAB8964

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Citations (3)

Product Documents

Recombinant Monoclonal Antibody.

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

Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human

Cited:

Mouse

Applications

Validated:

Western Blot, Immunocytochemistry

Cited:

Western Blot

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Clone # 1015B

View all p70 S6 Kinase Primary Antibodies »

Product Specifications

Immunogen

Phosphopeptide containing human p70S6K T229

Specificity

Detects human p70 S6 Kinase when phosphorylated at T229.

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for Human phospho-p70 S6 Kinase (T229) Antibody (1015B)

Detection of Human Phospho-p70 S6 Kinase (T229) by Western Blot.

Western blot shows lysates of MCF-7 human breast cancer cell line untreated (-) or treated (+) with 100 ng/mL Recombinant Human IGF-I (Catalog # 291-G1) for 1 hour. PVDF membrane was probed with 0.5 µg/mL of Rabbit Anti-Human Phospho-p70 S6 Kinase (T229) Monoclonal Antibody (Catalog # MAB8964) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). Specific bands were detected for Phospho-p70 S6 Kinase (T229) at approximately 70 and 85 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Phospho-p70 S6 Kinase (T229) in MCF‑7 Human Cell Line.

p70 S6 Kinase phosphorylated at T229 was detected in immersion fixed MCF-7 human breast cancer cell line treated with 100 ng/mL Recombinant Human IGF-I (Catalog # 291-G1) using Rabbit Anti-Human Phospho-p70 S6 Kinase (T229) Monoclonal Antibody (Catalog # MAB8964) at 2.5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Applications for Human phospho-p70 S6 Kinase (T229) Antibody (1015B)

Application

Recommended Usage

Immunocytochemistry

2.5-25 µg/mL
Sample: Immersion fixed MCF‑7 human breast cancer cell line treated with 100 ng/mL Recombinant Human IGF-I (Catalog # 291-G1)

Western Blot

0.5 µg/mL
Sample: MCF‑7 human breast cancer cell line treated with Recombinant Human IGF‑I (Catalog # 291-G1)

Formulation, Preparation, and Storage

Purification

Protein A or G purified from cell culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

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.

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: p70 S6 Kinase

p70 S6 Kinase (p70S6K) is responsible for the phosphorylation of 40S ribosomal protein S6 and is ubiquitously expressed in human adult tissues (1). p70S6K is activated by serum stimulation and this activation is inhibited by wortmannin and rapamycin. p70S6K activity undergoes changes in the cell cycle and increases 20-fold in G1 cells released from G0 (2). p70S6K activation requires sequential phosphorylations at proline-directed residues in the putative autoinhibitory pseudosubstrate domain, as well as T389, a site phosphorylated by Phosphoinositide-Dependent Kinase 1 (PDK1).