Cys-Protein-A/G/L

Cys-Protein A/G/L Recombinant
Cat. No.
BT8777
Source
Escherichia Coli.
Synonyms
Appearance
Sterile Filtered White lyophilized (freeze-dried) powder.
Purity
Greater than 95.0% as determined by:
(a) Analysis by SEC-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.
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Description

Recombinant Protein-A/G/L produced in E.Coli is a single non-glycosylated polypeptide chain fused with a Cys at N-terminus. Protein- A/G/L is comprised of 5 IgG-binding regions of Protein A (E-D-A-B-C), 2 of protein G (C1-C3) and 5 of Protein L (B1-B2-B3-B4-B5) containing 806 amino acids in total and having a molecular mass of 89.3kDa. Cell wall binding region, cell membrane binding region and albumin binding region have been eliminated from the recombinant Protein- A/G/L to guarantee the maximum specific IgG binding.

Product Specs

Introduction
Recombinant Protein A/G/L is a genetically engineered protein that combines the IgG binding characteristics of Protein A, Protein G, and Protein L. This fusion protein consists of five Ig-binding regions from Protein L (B1-B2-B3-B4-B5), five IgG binding domains from Protein A (E-D-A-B-C), and two Ig-binding regions from Protein G (C1-C3). Its ability to bind to IgG from various species, including humans, mice, rats, cows, goats, sheep, rabbits, guinea pigs, pigs, dogs, and cats, makes it ideal for purifying monoclonal or polyclonal IgG antibodies.
Description
Produced in E. coli, Recombinant Protein-A/G/L is a single, non-glycosylated polypeptide chain with a cysteine residue added at the N-terminus. This 89.3kDa protein comprises 806 amino acids, encompassing five IgG-binding regions from Protein A (E-D-A-B-C), two from Protein G (C1-C3), and five from Protein L (B1-B2-B3-B4-B5). The cell wall binding region, cell membrane binding region, and albumin binding region have been removed to ensure maximum specificity for IgG binding.
Physical Appearance
Sterile Filtered White Lyophilized Powder
Formulation
Protein-A/G/L is lyophilized without the addition of any excipients or stabilizers.
Solubility
To reconstitute lyophilized Protein-A/G/L, it is recommended to dissolve it in sterile, 18M-cm H2O at a minimum concentration of 0.1mg/ml. This solution can be further diluted with other aqueous solutions as needed.
Stability
Lyophilized Protein-A/G/L remains stable at room temperature for up to three weeks; however, it is recommended to store it desiccated at a temperature below -18°C. After reconstitution, Protein-A/G/L should be stored at 4°C for no longer than 2-7 days. For long-term storage, it is recommended to freeze it below -18°C after adding a carrier protein such as 0.1% HSA or BSA. Avoid repeated freeze-thaw cycles.
Purity
The purity of Protein-A/G/L is greater than 95.0% as determined by Size Exclusion Chromatography-High Performance Liquid Chromatography (SEC-HPLC) and Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis (SDS-PAGE) analysis.
Source
Escherichia Coli.
Amino Acid Sequence
CNAAQHDEAQ QNAFYQVLNM PNLNADQRNG FIQSLKDDPS QSANVLGEAQ KLNDSQAPKA DAQQNNFNKD QQSAFYEILN MPNLNEAQRN GFIQSLKDDP SQSTNVLGEA KKLNESQAPK ADNNFNKEQQ NAFYEILNMP NLNEEQRNGF IQSLKDDPSQ SANLLSEAKK LNESQAPKAD NKFNKEQQNA FYEILHLPNL NEEQRNGFIQ SLKDDPSQSA NLLAEAKKLN DAQAPKADNK FNKEQQNAFY EILHLPNLTE EQRNGFIQSL KDDPSVSKEI LAEAKKLNDA QAPKEEDSLE GSGSGTYKLI LNGKTLKGET TTEAVDAATA EKVFKQYAND NGVDGEWTYD DATKTFTVTE KPEVIDASEL TPAVTTYKLV INGKTLKGET TTKAVDAETA EKAFKQYAND NGVDGVWTYD DATKTFTVTE EPRARPGSGS GKEETPETPE TDSEEEVTIK ANLIFANGST QTAEFKGTFE KATSEAYAYA DTLKKDNGEY TVDVADKGYT LNIKFAGKEK TPEEPKEEVT IKANLIYADG KTQTAEFKGT FEEATAEAYR YADALKKDNG EYTVDVADKG YTLNIKFAGK EKTPEEPKEE VTIKANLIYA DGKTQTAEFK GTFEEATAEA YRYADLLAKE NGKYTVDVAD KGYTLNIKFA GKEKTPEEPK EEVTIKANLI YADGKTQTAE FKGTFAEATA EAYRYADLLA KENGKYTADL EDGGYTINIR FAGKKVDEKP EEKEQVTIKE NIYFEDGTVQ TATFKGTFAE ATAEAYRYAD LLSKEHGKYT ADLEDGGYTI NIRFAG.

Product Science Overview

Structure and Composition

The recombinant Cys-Protein A/G/L consists of:

  • Five IgG-binding regions of Protein A: Protein A is derived from the bacterium Staphylococcus aureus and is known for its ability to bind to the Fc region of IgG molecules.
  • Two IgG-binding regions of Protein G: Protein G is derived from Streptococcus species and also binds to the Fc region of IgG, but with a slightly different binding profile compared to Protein A.
  • Five Ig-binding regions of Protein L: Protein L is derived from the bacterium Peptostreptococcus magnus and binds to the light chains of IgG, providing an additional binding mechanism.

The fusion protein is engineered to remove non-specific binding regions, such as cell wall binding regions, cell membrane binding regions, and albumin binding regions, to ensure maximum specific IgG binding .

Production and Purity

Cys-Protein A/G/L is produced in Escherichia coli (E. coli) and is a single non-glycosylated polypeptide chain containing 806 amino acids with a cysteine (Cys) residue at the N-terminus. The molecular weight of the recombinant protein is approximately 89.3 kDa . The protein is typically purified to a purity level of greater than 95% as determined by SDS-PAGE and HPLC analyses .

Applications

The primary application of Cys-Protein A/G/L is in the purification of IgG antibodies. The recombinant protein’s ability to bind to a wide range of IgG molecules from different species makes it an ideal tool for antibody purification in research and diagnostic applications . The removal of non-specific binding regions ensures high specificity and efficiency in IgG binding, making it a valuable reagent in immunoprecipitation, affinity chromatography, and other antibody-based assays .

Stability and Storage

Cys-Protein A/G/L is typically supplied as a lyophilized (freeze-dried) powder, which should be stored at -20°C to -70°C to maintain its stability. After reconstitution in distilled water or saline, the protein can be stored under sterile conditions at 2 to 8°C for up to one month or at -20°C to -70°C for up to three months .

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