Ku P70/P80 Human

Ku P70/P80 Human Recombinant
Cat. No.
BT19178
Source
Sf9 insect cells.
Synonyms
ATP-dependent DNA helicase 2 subunit 1, ATP-dependent DNA helicase II 70 kDa subunit, Lupus Ku autoantigen protein p70, Ku70, 70 kDa subunit of Ku antigen, Thyroid-lupus autoantigen, TLAA, CTC box-binding factor 75 kDa subunit, CTCBF, CTC75, DNA-repair protein XRCC6, G22P1, XRCC6, ML8.
Appearance
Sterile Filtered clear solution.
Purity
Greater than 95% as determined by SDS-PAGE.
Usage
Prospec's products are furnished for LABORATORY RESEARCH USE ONLY. The product may not be used as drugs, agricultural or pesticidal products, food additives or household chemicals.
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Description

KuP70/P80 Human Recombinant produced in SF9 insect cells is a glycosylated, polypeptide chain having a molecular mass of 70,638 Dalton for the p70 subunit and 83,528 Dalton for the p80 subunit.
Ku (P70/P80) is expressed with a -6xHis tag and purified by proprietary chromatographic techniques.

Product Specs

Introduction
The Ku antigen comprises two polypeptides, p70 (70 kDa) and p80 (80 kDa), sometimes referred to as p86. This Ku heterodimer confers DNA-binding and regulatory capabilities to the DNA-dependent protein kinase, a crucial enzyme for DNA recombination in eukaryotic cells. The Ku protein plays a critical role in various nuclear processes, including DNA repair, V(D)J recombination, telomere maintenance, and the regulation of specific gene transcription. Notably, antibodies targeting Ku were first identified in individuals with scleroderma-polymyositis overlap syndrome (observed in up to 20% of patients). These antibodies have also been detected in approximately 20% of patients with primary pulmonary hypertension (often a complication of the overlap syndrome), 5-10% of patients with systemic lupus erythematosus (SLE) and mixed connective tissue disease (MCTD), and up to 20% of patients with primary Sjögren's syndrome. When using HEp-2 cells in indirect immunofluorescence (IIF) assays, Ku-positive sera exhibit a distinctive dense, fine granular pattern with nucleolar staining. However, this staining pattern is not exclusive to anti-Ku antibodies. Detecting Ku antibodies via Western blot can lead to false-negative results, as the primary reactive epitope on the p70/p80 heterodimer is conformation-dependent.
Description
KuP70/P80 Human Recombinant, produced in SF9 insect cells, is a glycosylated polypeptide chain. The p70 subunit has a molecular mass of 70,638 Daltons, while the p80 subunit has a molecular mass of 83,528 Daltons. This Ku (P70/P80) recombinant protein is expressed with a 6xHis tag and purified using proprietary chromatographic techniques.
Physical Appearance
The product appears as a clear solution that has undergone sterile filtration.
Formulation
KuP70/P80 is supplied in a buffer solution containing 16mM HEPES (pH 8), 160mM NaCl, and 20% glycerol.
Immunological Functions
This product serves two primary immunological functions: 1) It binds to IgG-type human auto-antibodies. 2) It acts as a standard in ELISA tests, enabling checkerboard analysis of positive and negative sera panels, including internationally recognized reference sera from the Centers for Disease Control and Prevention (CDC).
Coating Concentration
The recommended coating concentration for this product ranges from 0.2 to 0.6 µg/ml. The optimal concentration may vary depending on the specific ELISA plate and coating buffer used. This product is suitable for biotinylation and iodination procedures.
Stability
For short-term storage (2-4 weeks), keep the product refrigerated at 4°C. For extended storage, freeze the product at -20°C. Repeated freezing and thawing should be avoided.
Purity
The purity of this product is greater than 95%, as determined by SDS-PAGE analysis.
Synonyms
ATP-dependent DNA helicase 2 subunit 1, ATP-dependent DNA helicase II 70 kDa subunit, Lupus Ku autoantigen protein p70, Ku70, 70 kDa subunit of Ku antigen, Thyroid-lupus autoantigen, TLAA, CTC box-binding factor 75 kDa subunit, CTCBF, CTC75, DNA-repair protein XRCC6, G22P1, XRCC6, ML8.
Source
Sf9 insect cells.

Product Science Overview

Introduction

Ku P70/P80, also known as the Ku heterodimer, is a crucial protein complex involved in DNA repair mechanisms. This complex consists of two subunits: Ku70 (P70) and Ku80 (P80). The recombinant form of this protein, produced in various expression systems, is widely used in research to study its functions and applications.

Historical Background

The Ku70/80 heterodimer was first identified from the sera of patients with systemic lupus erythematosus (SLE), an autoimmune disease where the immune system mistakenly attacks healthy tissue . The Ku proteins were later found to play a significant role in DNA repair, particularly in the non-homologous end joining (NHEJ) pathway, which is essential for repairing double-strand breaks in DNA .

Structure and Expression

The Ku heterodimer is composed of two polypeptides: a 70 kDa subunit (Ku70) and an 80 kDa subunit (Ku80). The recombinant form of Ku P70/P80 is often produced in insect cells (Sf9) and is glycosylated, with molecular masses of approximately 70,638 Daltons for Ku70 and 83,528 Daltons for Ku80 . The recombinant protein is typically tagged with a His-tag for purification purposes .

Biological Functions

Ku P70/P80 plays a pivotal role in several cellular processes:

  1. DNA Repair: The Ku heterodimer binds to DNA ends with high affinity, protecting them from degradation and recruiting other proteins necessary for the NHEJ repair pathway .
  2. V(D)J Recombination: This process is crucial for the development of the immune system, allowing for the generation of diverse antibody repertoires .
  3. Telomere Maintenance: Ku proteins are involved in protecting telomeres, the ends of chromosomes, from degradation and fusion .
Clinical Significance

Mutations or deficiencies in Ku70 or Ku80 can lead to severe consequences, including increased sensitivity to radiation, defective DNA repair, and a higher risk of cancer . Studies have shown that downregulation or single nucleotide polymorphisms (SNPs) in these genes can contribute to various types of cancer, including breast, lung, and gastric cancers .

Applications

Recombinant Ku P70/P80 is used in various research applications, including:

  1. ELISA: To detect auto-antibodies in patient sera .
  2. Western Blotting: For the detection of Ku proteins using specific antibodies .
  3. Biochemical Studies: To understand the mechanisms of DNA repair and the role of Ku proteins in maintaining genomic stability .

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