Protein-A/G/L-Cys
Protein A/G/L-Cys Recombinant
Cat. No.
BT9560
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.
(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 C-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 comprises 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 broad binding profile makes it highly effective for purifying monoclonal and polyclonal IgG antibodies from various species, including humans, mice, rats, cows, goats, sheep, rabbits, guinea pigs, pigs, dogs, and cats.
Description
Recombinant Protein-A/G/L, expressed in E. coli, is a single, non-glycosylated polypeptide chain with a C-terminal Cys tag. This 89.3 kDa protein consists of 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 recombinant Protein-A/G/L is engineered for maximum IgG binding specificity by eliminating cell wall binding, cell membrane binding, and albumin binding regions.
Physical Appearance
Sterile Filtered White lyophilized powder.
Formulation
Protein-A/G/L is lyophilized without the addition of any other substances.
Solubility
To reconstitute lyophilized Protein-A/G/L, it is recommended to dissolve the protein in sterile 18M-cm H2O to a concentration of at least 0.1mg/ml. This solution can be further diluted into other aqueous buffers or solutions as needed.
Stability
Lyophilized Protein-A/G/L remains stable at room temperature for up to 3 weeks. However, for extended storage, it should be kept desiccated at temperatures below -18°C. After reconstitution, Protein-A/G/L can be stored at 4°C for 2-7 days. For long-term storage, it is advisable to freeze the solution below -18°C after adding a carrier protein such as 0.1% HSA or BSA. Repeated freezing and thawing of the protein should be avoided.
Purity
The purity of this protein is greater than 95.0% as determined by: (a) Size-Exclusion High-Performance Liquid Chromatography (SEC-HPLC) and (b) Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis (SDS-PAGE).
Source
Escherichia Coli.
Amino Acid Sequence
NAAQHDEAQQ NAFYQVLNMP NLNADQRNGF IQSLKDDPSQ SANVLGEAQK LNDSQAPKAD AQQNNFNKDQ QSAFYEILNM PNLNEAQRNG FIQSLKDDPS QSTNVLGEAK KLNESQAPKA DNNFNKEQQN AFYEILNMPN LNEEQRNGFI QSLKDDPSQS ANLLSEAKKL NESQAPKADN KFNKEQQNAF YEILHLPNLN EEQRNGFIQS LKDDPSQSAN LLAEAKKLND AQAPKADNKF NKEQQNAFYE ILHLPNLTEE QRNGFIQSLK DDPSVSKEIL AEAKKLNDAQ APKEEDSLEG SGSGTYKLIL NGKTLKGETT TEAVDAATAE KVFKQYANDN GVDGEWTYDD ATKTFTVTEK PEVIDASELT PAVTTYKLVI NGKTLKGETT TKAVDAETAE KAFKQYANDN GVDGVWTYDD ATKTFTVTEE PRARPGSGSG KEETPETPET DSEEEVTIKA NLIFANGSTQ TAEFKGTFEK ATSEAYAYAD TLKKDNGEYT VDVADKGYTL NIKFAGKEKT PEEPKEEVTI KANLIYADGK TQTAEFKGTF EEATAEAYRY ADALKKDNGE YTVDVADKGY TLNIKFAGKE KTPEEPKEEV TIKANLIYAD GKTQTAEFKG TFEEATAEAY RYADLLAKEN GKYTVDVADK GYTLNIKFAG KEKTPEEPKE EVTIKANLIY ADGKTQTAEF KGTFAEATAE AYRYADLLAK ENGKYTADLE DGGYTINIRF AGKKVDEKPE EKEQVTIKEN IYFEDGTVQT ATFKGTFAEA TAEAYRYADL LSKEHGKYTA DLEDGGYTIN IRFAGC.
Product Science Overview
Composition and Structure
Protein A/G/L-Cys Recombinant is composed of:
- Five IgG-binding regions of Protein A: Derived from Staphylococcus aureus segments E, D, A, B, and C.
- Two IgG-binding regions of Protein G: Derived from Streptococcus segments C1 and C3.
- Five IgG-binding regions of Protein L: Derived from Peptostreptococcus magnus segments B1, B2, B3, B4, and B5 .
The recombinant protein is a single non-glycosylated polypeptide chain containing 806 amino acids and has a molecular mass of approximately 89.3 kDa . It is fused with a cysteine (Cys) residue at the C-terminus to facilitate conjugation and immobilization .
Expression and Purification
Binding Specificity
The recombinant protein has been engineered to eliminate non-specific binding regions, such as cell wall binding, cell membrane binding, and albumin binding regions . This ensures maximum specific IgG binding. Protein A/G/L-Cys Recombinant can bind to IgG antibodies from a wide range of species, including human, mouse, rat, cow, goat, sheep, rabbit, guinea pig, pig, dog, and cat .
Applications
Protein A/G/L-Cys Recombinant is widely used in various applications, including:
Storage and Stability
The lyophilized protein is stable at room temperature for up to three weeks but should be stored desiccated below -18°C for long-term storage . Upon reconstitution, it should be stored at 4°C for short-term use (2-7 days) and below -18°C for long-term use. To prevent freeze-thaw cycles, it is recommended to add a carrier protein such as 0.1% human serum albumin (HSA) or bovine serum albumin (BSA) .
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