CD14 Human, CHO

CD14 Human Recombinant, CHO
Cat. No.
BT25617
Source
CHO-cells.
Synonyms
Monocyte differentiation antigen CD14, Myeloid cell-specific leucine-rich glycoprotein, CD14.
Appearance
Sterile Filtered White lyophilized (freeze-dried) powder.
Purity
Greater than 95.0% 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

The CD14 is produced from human CD14 transfected CHO-cells. Before transfection the complete human CD14-cDNA was amplified by PCR and cloned into expression vector p-POL-DHFR. The myeloid differentiation antigen CD14 acts as the major receptor for bacterial LPS. The dominant form of the recombinant wild type CD14 is the 50-kDa protein containing 335 amino acids and lacks the GPI-Anchor.

Product Specs

Introduction
CD14, also known as the lipopolysaccharide (LPS) receptor, is primarily found on monocytes and macrophages, with weak expression on neutrophils. It anchors to cells via glycosylphosphatidylinositol (GPI) and acts as a high-affinity receptor for LPS-LPS binding protein (LBP) complexes. Soluble CD14 can bind LPS, functioning as an agonist at physiological concentrations and an antagonist at higher concentrations in cell activation. CD14 has also been observed to bind apoptotic cells.
Description
Recombinant human CD14 is produced in CHO cells transfected with the full-length human CD14 cDNA, which was PCR amplified and cloned into the p-POL-DHFR expression vector. As the primary receptor for bacterial LPS, CD14 plays a crucial role in myeloid differentiation. The predominant form of this recombinant wild-type CD14 is a 50 kDa protein comprising 335 amino acids and lacking the GPI anchor.
Physical Appearance
Sterile white lyophilized powder.
Formulation
CD14 is lyophilized from a 1.0 mg/ml solution in phosphate-buffered saline (pH 7.2).
Solubility
Reconstitute the lyophilized CD14 in sterile 18 MΩ-cm H2O to a concentration of at least 100 µg/ml. Subsequent dilutions should be made using phosphate-buffered saline (PBS).
Stability
While lyophilized CD14 remains stable at room temperature for up to 3 weeks, it is recommended to store it desiccated at temperatures below -18°C. Reconstituted CD14 can be stored at 4°C for 2-7 days. For long-term storage, freeze at -18°C, ideally with the addition of a carrier protein (0.1% HSA or BSA). Avoid repeated freeze-thaw cycles.
Purity
Purity is determined to be greater than 95.0% by SDS-PAGE analysis.
Biological Activity
CD14 exhibits biological activity by inhibiting the binding of FITC-LPS (0.5 µg/ml) to 600,000 CD14+ CHO transfectants (measured by FACS) at concentrations up to 20 µg/ml.
Synonyms
Monocyte differentiation antigen CD14, Myeloid cell-specific leucine-rich glycoprotein, CD14.
Source
CHO-cells.

Product Science Overview

Introduction

CD14 is a glycoprotein that plays a crucial role in the immune system. It is primarily expressed on the surface of monocytes and macrophages. The recombinant form of human CD14, produced in Chinese Hamster Ovary (CHO) cells, is widely used in research and therapeutic applications due to its ability to mimic the natural protein’s functions.

Structure and Expression

CD14 is a 55 kDa cell surface glycoprotein. The human CD14 cDNA encodes a 375 amino acid precursor protein with a 19 amino acid signal peptide and a C-terminal hydrophobic region characteristic of glycosylphosphatidylinositol (GPI)-anchored proteins . The recombinant human CD14 protein produced in CHO cells is typically fused with a polyhistidine tag at the C-terminus to facilitate purification .

Biological Functions

CD14 acts as a co-receptor for the detection of bacterial lipopolysaccharide (LPS), a component of the outer membrane of Gram-negative bacteria. It enhances the LPS-stimulated secretion of pro-inflammatory cytokines such as IL-8 by immune cells . This function is critical for the activation of the innate immune response during bacterial infections.

Production and Purification

The recombinant human CD14 protein is produced by transfecting CHO cells with the complete human CD14 cDNA. The protein is then purified using techniques such as SDS-PAGE and HPLC to ensure high purity and low endotoxin levels . The lyophilized protein is typically stored at -20°C to -80°C to maintain stability and activity .

Applications

Recombinant human CD14 is used in various research applications, including:

  • Immunology Studies: Understanding the role of CD14 in the immune response.
  • Drug Development: Screening for potential therapeutic agents that modulate CD14 activity.
  • Diagnostic Assays: Developing assays to detect bacterial infections based on CD14’s interaction with LPS.

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