SPA Long

Staphylococcal Protein-A Long Form Recombinant
Cat. No.
BT10131
Source
Escherichia Coli.
Synonyms
Immunoglobulin G-binding protein A, IgG-binding protein A, Staphylococcal protein A, SPA.
Appearance
Sterile Filtered White lyophilized (freeze-dried) powder.
Purity
Greater than 97.0% as determined by:
(a) Analysis by HPLC.
(b) Analysis 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.
Shipped with Ice Packs
In Stock

Description

SPA Recombinant Long Form produced in E.Coli is a single non-glycosylated polypeptide chain. SPA Long Form is comprised of 5 IgG-binding domains E-D-A-B-C aligned in series containing 422 amino acids and having a molecular mass of 46.6kDa.

Product Specs

Introduction
Protein A, a natural component of the Staphylococcus aureus cell wall, is frequently used in its recombinant form for antibody purification. This engineered protein retains the five IgG-binding regions of native Protein A, making it highly effective for purifying both polyclonal and monoclonal IgG antibodies. Recombinant Protein A exhibits binding affinity for IgG1, IgG2, and IgG4 from human; IgG2a, IgG2b, and IgG3 from mouse; and IgG2c from rat. Additionally, it demonstrates binding capability for total IgG from various species, including rabbit, pig, dog, cat, and guinea pig.
Description

Produced in E. coli, SPA Recombinant Long Form is a single, non-glycosylated polypeptide chain. This form encompasses five IgG-binding domains (E-D-A-B-C) arranged sequentially, resulting in a protein composed of 422 amino acids with a molecular weight of 46.6kDa.

Physical Appearance
Sterile Filtered White lyophilized powder.
Formulation
SPA protein is lyophilized without any additional ingredients.
Solubility
To reconstitute lyophilized SPA, it is recommended to dissolve it in sterile 18MΩ-cm H2O to a minimum concentration of 0.1mg/ml. This solution can be further diluted in other aqueous solutions as needed.
Stability
While lyophilized SPA remains stable at room temperature for up to 3 weeks, it is recommended to store it desiccated at a temperature below -18°C for extended periods. After reconstitution, SPA should be stored at 4°C for 2-7 days. For long-term storage, freezing below -18°C is advised. The addition of a carrier protein, such as 0.1% HSA or BSA, is recommended for long-term storage.
Avoid repeated freeze-thaw cycles.
Purity
Purity exceeds 97.0% as determined by:
(a) High-performance liquid chromatography (HPLC) analysis.
(b) Sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis.
Synonyms
Immunoglobulin G-binding protein A, IgG-binding protein A, Staphylococcal protein A, SPA.
Source
Escherichia Coli.
Amino Acid Sequence
AQHDEAQQNA FYQVLNMPNL NADQRNGFIQ SLKDDPSQSA NVLGEAQKLN DSQAPKADAQ QNNFNKDQQS AFYEILNMPN LNEAQRNGFI QSLKDDPSQS TNVLGEAKKL NESQAPKADN NFNKEQQNAF YEILNMPNLN EEQRNGFIQS LKDDPSQSAN LLSEAKKLNE SQAPKADNKF NKEQQNAFYE ILHLPNLNEE QRNGFIQSLK DDPSQSANLL AEAKKLNDAQ APKADNKFNK EQQNAFYEIL HLPNLTEEQR NGFIQSLKDD PSVSKEILAE AKKLNDAQAP KEEDNKKPGK EDGNKPGKED GNKPGKEDNK KPGKEDGNKP GKEDNNKPGK EDGNKPGKED NNKPGKEDGN KPGKEDGNKP GKEDGNGVHV VKPGDTVNDI AKANGTTADK IAADNKLADK NMIKPGQELV VD.

Product Science Overview

Introduction

Staphylococcal Protein-A (SpA) is a cell wall protein found in the bacterium Staphylococcus aureus. It is known for its ability to bind to the Fc region of immunoglobulins, particularly Immunoglobulin G (IgG). This binding property makes SpA a valuable tool in various biochemical and immunological applications, including antibody purification and immunoprecipitation.

Structure and Composition

The long form of recombinant Staphylococcal Protein-A is a non-glycosylated polypeptide chain produced in Escherichia coli (E. coli). It consists of five IgG-binding domains, labeled E, D, A, B, and C, aligned in series. This recombinant protein comprises 422 amino acids and has a molecular mass of approximately 46.6 kDa . The long form is designed to enhance its binding capacity and stability, making it suitable for various laboratory and industrial applications.

Preparation Methods

The recombinant long form of Staphylococcal Protein-A is typically produced using E. coli expression systems. The gene encoding the protein is cloned into an expression vector, which is then introduced into E. coli cells. The bacteria are cultured under conditions that promote the expression of the recombinant protein. After sufficient growth, the cells are harvested, and the protein is extracted and purified using techniques such as affinity chromatography. The final product is often lyophilized (freeze-dried) to ensure stability and ease of storage .

Applications and Uses

Staphylococcal Protein-A is widely used in the field of biotechnology and immunology. Its primary application is in the purification of antibodies. The protein’s ability to bind to the Fc region of IgG allows it to be used in affinity chromatography columns to isolate antibodies from complex mixtures. Additionally, SpA is employed in immunoprecipitation assays to pull down specific proteins from cell lysates using antibodies. It is also used in the production of affinity resins for various immunological assays .

Stability and Storage

The lyophilized form of Staphylococcal Protein-A is stable at room temperature for up to three weeks. For long-term storage, it is recommended to keep the protein desiccated at temperatures below -18°C. Upon reconstitution, the protein should be stored at 4°C for short-term use (2-7 days) and below -18°C for long-term use. To prevent degradation, it is advisable to add a carrier protein, such as human serum albumin (HSA) or bovine serum albumin (BSA), and avoid repeated freeze-thaw cycles .

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