Antibody purification entails isolating antibodies from sources such as culture supernatant of hybridoma cell line, ascites fluid, or serum. The level of purification necessary to obtain a usable antibody depends on the antibody's intended use. Supported by our advanced technology platform and experienced scientists, Creative Biolabs offers high-quality and various kinds of antibody purification and characterization products to accelerate your programs and meet your study milestones.
Antibodies are classified into different isotypes or fragments based on their structure. Antibody characterization is necessary for choosing an appropriate purification method. Antibody purification is a critical step in preparing antibodies that specific to target antigens. The methods used can vary significantly in terms of complexity, yield, and purity, depending on the source of the antibodies and the desired application. Antibodies are most frequently purified using affinity purification techniques.
Because antibodies have predictable structures, including relatively invariant domains, certain protein ligands are capable of binding to the antibody. Protein A and G are two bacterial proteins that can bind specifically to the Fc region of antibodies, making these proteins useful for antibody purification. This method provides high yield and is widely used for purifying IgG antibodies from various species. In addition, the genetic engineering form of Protein A and G, known as Protein A/G, which is also excellent for IgG antibodies purification. Protein L is another valuable tool in the field of antibody purification due to its unique binding properties to kappa light chains. The features of these IgG-binding proteins are listed in Table 1.
Table 1 Characteristics of Proteins A, G, A/G, and L
Protein A | Protein G | Protein A/G | Protein L | |
Production Source | E. coli | E. coli | E. coli | E. coli |
Molecular Weight | 44.6 kDa | 21.6 kDa | 50.4 kDa | 35.8 kDa |
Number of Binding Sites for IgG | 4 | 2 | 6 | 4 |
Binding pH | 8.2 | 5 | 5-8.2 | 7.5 |
Binds Site | Fc | Fc | Fc | VLK |
For different species or antibody subclasses the interactions between various proteins and antibodies are not the same. Determine the best affinity protein for your application by consulting Table 2 and Table 3. Creative Biolabs provides different easy to use and high quality antibody characterization kits for preparing antibody fragments, identifying antibody isotypes and subclasses, and measuring antibody titers.
Table 2 Binding characteristics of Proteins A, G, A/G, and L to different species IgG
Protein A | Protein G | Protein A/G | Protein L | |
Human IgG | +++ | +++ | +++ | +++ |
Mouse IgG | +++ | +++ | +++ | +++ |
Rabbit IgG | +++ | +++ | +++ | + |
Goat IgG | + | +++ | +++ | - |
Rat IgG | + | ++ | ++ | +++ |
Hamster IgG | ++ | ++ | ++ | +++ |
Donkey IgG | ++ | +++ | +++ | / |
Dog IgG | +++ | + | +++ | / |
Cat IgG | +++ | + | +++ | / |
Pig IgG | +++ | + | +++ | +++ |
Horse IgG | + | +++ | +++ | / |
Note:+++ means strong binding; ++ means medium binding; + means weak binding; - means no binding; / means no sufficient information.
Table 3 Binding characteristics of Proteins A, G, A/G, and L to different antibody subclasses
Protein A | Protein G | Protein A/G | Protein L | |
Human IgG1 | +++ | +++ | +++ | +++ |
Human IgG2 | +++ | +++ | +++ | +++ |
Human IgG3 | + | +++ | +++ | +++ |
Human IgG4 | +++ | +++ | +++ | +++ |
Human Fab | + | + | + | +++ |
Human ScFv | + | - | + | +++ |
Mouse IgG1 | + | ++ | ++ | +++ |
Mouse IgG2a | +++ | +++ | +++ | +++ |
Mouse IgG2b | +++ | +++ | +++ | +++ |
Mouse IgG3 | +++ | +++ | +++ | +++ |
Rat IgG1 | + | ++ | ++ | +++ |
Rat IgG2a | - | +++ | +++ | +++ |
Rat IgG2b | - | + | + | +++ |
Rat IgG2c | +++ | +++ | +++ | +++ |
Note:+++ means strong binding; ++ means medium binding; + means weak binding; - means no binding.
Optimal binding conditions vary for each protein. Therefore, each immobilized affinity purification product is supplied with a separate IgG binding buffer. Using our IgG elution buffer can make the elution of IgG sharp and thorough. Our binding and elution buffers are optimized to provide the highest IgG binding and elution efficiencies using immobilized Protein A, Protein G, and Protein A/G.
Although Protein A, G, A/G, and L can satisfy the purification requirements of most species and subtypes of IgG, there are still other types of antibodies that require other purification methods, such as IgA, IgM, and IgY.
Human IgA is most commonly purified by jacalin. Jacalin's ability to bind to human IgA makes it useful in various immunological assays and research. Jacalin immobilized on agarose can be used to purify human serum or secretory IgA1, and can also be used to remove contaminating IgA from IgG samples. We offer immobilized jacalin on cross-linked 6% agarose (CAT#: Glyco-061CL).
IgM is a high molecular mass glycoprotein. It generally has a pentameric structure in the serum of mammals. We devised a convenient to use immobilized MBP (Mannan Binding Protein) system, complete with a buffer, for purifying IgM. This method is particularly effective for isolating mouse IgM from ascites fluid.
IgY is a type of immunoglobulin found in birds, reptiles, and amphibians. In chickens, IgY is the primary antibody present in egg yolk. Fc-binding proteins such as Protein A and G do not bind IgY so that it is difficult to purify. Our chicken IgY purification Kit was developed for efficient purification of IgY from egg yolk.
Product Type | Product Name | Cat |
Protein A | Protein A Spin Columns (0.2 mL) | VS-0824-WK1 |
Protein A Spin Columns (1 mL) | VS-0824-WK2 | |
Protein A Spin Columns (5 mL) | VS-0824-WK3 | |
Recombinant Protein A Agarose | VS-0824-WK6 | |
Protein G | Protein G Spin Columns (0.2 mL) | VS-0824-WK7 |
Protein G Spin Columns (1 mL) | VS-0824-WK8 | |
Protein G Spin Columns (5 mL) | VS-0824-WK9 | |
Protein G Agarose | VS-0824-WK12 | |
Protein A/G | Protein A/G Spin Columns (0.2 mL) | VS-0824-WK13 |
Protein A/G Spin Columns (1 mL) | VS-0824-WK14 | |
Protein A/G Spin Columns (5 mL) | VS-0824-WK15 | |
Protein L | Protein L Spin Columns (0.2 mL) | VS-0824-WK18 |
Protein L Spin Columns (1 mL) | VS-0824-WK19 | |
Protein L Spin Columns (5 mL) | VS-0824-WK20 | |
Antibody Purification Kit | Protein A Spin Kit (0.2 mL) | VS-0824-WK4 |
Protein A Spin Kit (1 mL) | VS-0824-WK5 | |
Protein G Spin Kit (0.2 mL) | VS-0824-WK10 | |
Protein G Spin Kit (1 mL) | VS-0824-WK11 | |
Protein A/G Spin Kit (0.2 mL) | VS-0824-WK16 | |
Protein A/G Spin Kit (1 mL) | VS-0824-WK17 | |
Protein L Spin Kit (0.2 mL) | VS-0824-WK21 | |
Protein L Spin Kit (1 mL) | VS-0824-WK22 | |
IgM Purification Kit | VS-0824-WK23 | |
Chicken IgY Purification Kit | VS-0824-WK24 | |
Antibody Purification Buffer | Protein A IgG Binding Buffer | VS-0824-WK25 |
Protein G IgG Binding Buffer | VS-0824-WK26 | |
Protein A/G IgG Binding Buffer | VS-0824-WK27 | |
IgG Elution Buffer | VS-0824-WK28 |
Product Type | Product Name | Cat |
Antibody Fragment Preparation Kit | Fab Fragment Preparation Kit | VS-0924-WK1 |
F(ab')2 Fragment Preparation Kit | VS-0924-WK2 | |
Mouse IgG1 Fab and F(ab')2 Fragment Preparation Kit | VS-0924-WK3 | |
Antibody Isotyping Kit | Mouse Immunoglobulin Isotyping ELISA Kit | VS-0924-WK4 |
Rat Immunoglobulin Isotyping ELISA Kit | VS-0924-WK5 | |
Rapid Mouse Antibody Isotyping Kit, Dipstick-bassed | VS-0924-WK6 | |
Rapid Rat Antibody Isotyping Kit, Dipstick-based | VS-0924-WK7 | |
Rapid Human Antibody Isotyping Kit, Dipstick-based | VS-0924-WK8 | |
Rapid Mouse Antibody Isotyping Kit, Cassette-based | VS-0924-WK9 | |
Rapid Mouse Antibody Kappa and Lambda Isotyping Kit, Cassette-based | VS-0924-WK10 | |
Antibody Titer Assay Kit | Mouse IgG Titer Quantitation Assay Kit | VS-0924-WK11 |
Rabbit IgG Titer Quantitation Assay Kit | VS-0924-WK12 | |
Human IgG (H+L) Titer Quantitation Assay Kit | VS-0924-WK13 | |
Human IgG (Gamma chain) Titer Quantitation Assay Kit | VS-0924-WK14 | |
Human IgM Titer Quantitation Assay Kit | VS-0924-WK15 |
Creative Biolabs is committed to providing high-quality antibody purification and characterization products for global customers. With advanced technology platforms and professionals, we can provide custom various antibody purification products to meet our customers' needs. For more detailed information, please feel free to contact us.
For Research Use Only. Not For Clinical Use.