Protein Cys-A/G
Protein Cys-A/G Recombinant
Cat. No.
BT9203
Source
Escherichia Coli.
Synonyms
Appearance
Sterile Filtered White lyophilized (freeze-dried) powder.
Purity
Greater than 96.0% as determined by:
(a) Analysis by RP-HPLC.
(b) Analysis by SDS-PAGE.
(a) Analysis by RP-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
Protein-A/G Recombinant produced in E.Coli is a single non-glycosylated polypeptide chain fused with a Cys at N-terminus. Protein-A/G is comprised of 5 IgG-binding regions of protein A (E-D-A-B-C) and 2 of protein G (C1-C3) containing 430 amino acids in total and having a molecular mass of 47.8kDa. Cell wall binding region, cell membrane binding region and albumin binding region have been eliminated from the recombinant Protein A/G to guarantee the maximum specific IgG binding.
Product Specs
Introduction
Recombinant Protein A/G is a genetically engineered protein that combines seven IgG-binding domains from Protein A and Protein G (EDABC-C1C3). This design leverages the binding domains of both proteins, resulting in a broader binding capacity compared to using either protein alone. The Protein A portion is derived from Staphylococcus aureus segments E, D, A, B, and C, while the Protein G portion comes from Streptococcus segments C1 and C3. This characteristic makes recombinant Protein A/G highly effective for purifying both monoclonal and polyclonal IgG antibodies. Notably, it exhibits binding affinity for various human, mouse, and rat IgG subclasses, including human IgG1, IgG2, IgG3, IgG4; mouse IgG2a, IgG2b, IgG3; and rat IgG2a, IgG2c. Moreover, Protein A/G demonstrates binding to total IgG from diverse species such as cow, goat, sheep, horse, rabbit, guinea pig, pig, dog, and cat.
Description
Protein-A/G Recombinant, produced in E. coli, is a single, non-glycosylated polypeptide chain with a cysteine residue added at the N-terminus. This recombinant protein consists of 430 amino acids, encompassing five IgG-binding regions from protein A (E-D-A-B-C) and two from protein G (C1-C3), resulting in a molecular mass of 47.8 kDa. The design specifically excludes the cell wall binding region, cell membrane binding region, and albumin binding region to ensure maximum specificity for IgG binding.
Physical Appearance
Sterile Filtered White lyophilized powder.
Formulation
Protein-A/G is lyophilized without the addition of any other substances.
Solubility
To reconstitute the lyophilized Protein-A/G, it is recommended to dissolve it in sterile 18M-cm H2O at a concentration of at least 0.1 mg/ml. This solution can be further diluted in other aqueous solutions as needed.
Stability
Lyophilized Protein-A/G remains stable at room temperature for up to 3 weeks; however, for long-term storage, it is recommended to store it desiccated at a temperature below -18°C. Once reconstituted, Protein-A/G should be stored at 4°C for no more than 2-7 days. For extended storage, freezing at -18°C is advised. To preserve protein stability during long-term storage, the addition of a carrier protein such as HSA or BSA (0.1%) is recommended. It's important to avoid repeated freeze-thaw cycles to maintain protein integrity.
Purity
The purity of Protein-A/G is greater than 96.0%, as determined by two methods: (a) Reverse-Phase High-Performance Liquid Chromatography (RP-HPLC) analysis, and (b) 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.
Product Science Overview
Structure and Composition
The recombinant Protein A/G is composed of:
- Five IgG-binding regions of Protein A: These regions are derived from the segments E, D, A, B, and C of the Protein A from Staphylococcus aureus .
- Two IgG-binding regions of Protein G: These regions are derived from the segments C1 and C3 of the Protein G from Streptococcus .
The recombinant protein is produced in Escherichia coli and is typically available as a sterile filtered, white lyophilized (freeze-dried) powder .
Binding Capacity
Protein A/G has a broader binding capacity than either Protein A or Protein G alone. It binds to various human, mouse, and rat IgG subclasses, including:
- Human IgG1, IgG2, IgG3, IgG4
- Mouse IgG2a, IgG2b, IgG3
- Rat IgG2a, IgG2c
Additionally, it binds to total IgG from other species such as cow, goat, sheep, horse, rabbit, guinea pig, pig, dog, and cat .
Applications
The recombinant Protein A/G is ideal for the purification of monoclonal or polyclonal IgG antibodies. It is commonly used in:
- Affinity chromatography: For the purification of antibodies from various species.
- Immunoprecipitation: To isolate specific proteins from complex mixtures.
- Western blotting: As a detection reagent for IgG antibodies.
Stability and Storage
The lyophilized Protein A/G is stable at room temperature for up to three weeks. For long-term storage, it should be kept desiccated below -18°C. Upon reconstitution, it should be stored at 4°C for short-term use (2-7 days) and below -18°C for long-term use. It is recommended to add a carrier protein (0.1% HSA or BSA) to prevent freeze-thaw cycles .
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