Antibody purification is the first step in screening serum, ascites fluid or cell culture supernatant samples for a target antibody titer and helps to eliminate all other proteins from the sample. Once purified, the antibodies are typically screened by ELISA using the antigen of interest to selectively bind target-specific antibodies. This method allows researchers to determine which samples are producing the highest titer of target antibody and evaluate binding conditions. Thermo Scientific Pierce 96-well Spin Plates packed with antibody purification resin enable researchers to rapidly screen numerous samples for the presence of a target antibody and makes the antibody screening process faster.
The following Thermo Scientific Pierce 96-well Spin Plates are available for antibody purification and screening:
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Antibody Production and Purification Handbook
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Protein A Spin Plate for IgG Screening
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Protein A has long been used to purify mammalian antibodies and binds mammalian IgG's through the Fc region of the antibody with species-specific variability. Protein A is commonly used to purify humanized antibodies. Because Protein A binds IgG's specifically, sample pretreatment is typically not required.
When used in 96-well plates, 50µL of immobilized Protein A can process samples of 10 to 100µL and purify up to 0.6mg of human IgG per well. The "bind, wash and elute" method common for Protein A resins is effective for purifying IgG from ascites, cell culture and serum samples. Antibodies purified using the Thermo Scientific Pierce Protein A Spin Plates are typically eluted in a low-pH buffer that can be neutralized with alkaline Tris buffer or processed by buffer exchange using a product like the 96-well Zeba Spin Desalting Plates (Part No. 89807). Protein A Spin Plates can be used for high-throughput immunoprecipitation assays.
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96-well Filter Spin Plate Specifications and Applications
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| Technical Specifications for Thermo Scientific Pierce Protein A Spin Plates |
| Resin |
crosslinked 6% agarose |
| Ligand |
Protein A |
| Resin bed volume |
50µL |
| Binding capacity |
0.6mg human IgG/well |
| Sample volume per well |
10 to 100µL |
| Assay time |
1 hour |
| Coefficient of variation (CV %) of protein yield |
<20% |
| Centrifugation speed |
1000 x g |
| Storage buffer |
ultrapure water with 0.02% sodium azide |
| Storage temperature |
4°C |
| Suggested balance plate |
use Part No. 45205 |
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Pierce Protein A Agarose Resins, Columns and Plates
96-well Filter Collection Plates
96-well Spin Balance Plate
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Purification of human IgG from human serum using Thermo Scientific Protein A Spin Plate Kits. Human serum (40µL) was purified using one well of the Protein A Plate according to the product instructions. The collected samples and controls were prepared in Reducing Sample Buffer, separated by Tris-glycine SDS-PAGE and stained with Imperial Protein Stain (Part No. 24615). M = MW marker, I = human IgG, S = human serum, F = flow-through, E = eluate. |
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Protein G Spin Plate for IgG Screening
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Like Protein A, Protein G binds mammalian IgG's through the Fc region of the antibody with species-specific variability. More expensive, Protein G is typically used to purify monoclonal and polyclonal IgG's that do not bind well to Protein A.
When used in 96-well plates, 50µL of immobilized Protein G resin can process samples of 20 to 100µL and purify up to 0.5mg of human IgG per well. The "bind, wash and elute" method common for Protein G resins is effective for purifying IgG from ascites, cell culture and serum samples. Antibodies purified using the Thermo Scientific Pierce Protein G Spin Plates are typically eluted in a low-pH buffer that can be neutralized with alkaline Tris buffer or processed by buffer exchange using a product like the 96-well Zeba Spin Desalting Plates (Part No. 89807). Protein G Spin Plates can be used for high-throughput immunoprecipitation assays.
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Learn more...
96-well Filter Spin Plate Specifications and Applications
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| Technical Specifications for Thermo Scientific Pierce Protein G Spin Plates |
| Resin |
crosslinked 6% agarose |
| Ligand |
Protein G |
| Resin bed volume |
50µL |
| Binding capacity |
0.5mg human IgG/well |
| Sample volume per well |
20 to 100µL |
| Assay time |
1 hour |
| Coefficient of variation (CV %) of protein yield |
<20% |
| Centrifugation speed |
1000 x g |
| Storage buffer |
ultrapure water with 0.02% sodium azide |
| Storage temperature |
4°C |
| Suggested balance plate |
use Part No. 45205 |
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View Products...
Pierce Protein G Agarose Resins, Columns and Plates
96-well Filter Collection Plates
96-well Spin Balance Plate
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Purification of human IgG from human serum using Thermo Scientific Protein G Spin Plate Kits. Human serum (40µL) was purified using one well of the Protein G Plate according to the product instructions. The collected samples and controls were prepared in Reducing Sample Buffer, separated by Tris-glycine SDS-PAGE and stained with Imperial Protein Stain (Part No. 24615). M = MW marker, I = human IgG, S = human serum, F = flow-through, E = eluate. |
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Melon Gel Spin Plate for IgG Screening
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A faster antibody purification method is the ?negative selection? process used with Thermo Scientific Melon Gel. Melon Gel Resin can be used to process an antibody sample in as little as 15 minutes because there are no binding or elution steps. Unlike Protein A or G, Melon Gel is not species specific. Melon Gel binds non-relevant proteins, allowing the purified antibody to flow through the resin in physiological conditions suitable for storage and downstream applications. This flow-through purification process is especially useful for antibodies that lose their antigen-binding activity at low pH.
The robust 96-well Melon Gel purification procedure offers higher recovery and purity than bind-and-release methods. Each well of the 96-well plate contains 100µL of Melon Gel for processing samples of 10 to 100µL and will purify up to 1mg of IgG per well. The Melon Gel approach to antibody screening and purification requires cell culture supernatant, ascites and serum samples to be buffer exchanged or diluted in the required purification buffer before processing. This can easily be done using the 96-well Zeba Spin Desalting Plates (Part No. 89807).
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Learn more...
96-well Filter Spin Plate Specifications and Applications
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| Technical Specifications for Thermo Scientific Melon Gel Spin Plates |
| Resin |
Melon Gel Resin |
| Resin bed volume |
100µL |
| Purification capacity |
1mg human IgG/well |
| Sample volume per well |
10 to 100µL of cell culture supernatant, ascites fluid or serum |
| Assay time |
15 minutes |
| Coefficient of variation (CV %) of protein yield |
<10% |
| Centrifugation speed |
1000 x g |
| Storage buffer |
ultrapure water with 0.02% sodium azide |
| Storage temperature |
4°C |
| Suggested balance plate |
use Part No. 45205 |
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View Products...
Melon Gel Resins, Columns and Plates
96-well Filter Collection Plates
96-well Spin Balance Plate
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Thermo Scientific Melon Gel Spin Plate Purification of IgG from mouse ascites, culture media and serum. A. Ascites sample was reconstituted at 5mg/mL in water, treated with Ascites Conditioning Reagent (Part No. 45219), desalted and then 50µL of the sample was used for Melon Gel Spin Plate purification. B. Cell culture supernatant Ab110.1.1 was buffer exchanged into Melon Gel Purification Buffer and then 50µL used for Melon Gel Spin Plate purification. C. Human serum was diluted 1:10 with Melon Gel Purification Buffer and then 20µL was purified using one well of a Melon Gel Plate according to the product instructions. The unpurified and purified IgG samples were prepared in Reducing Sample Buffer, separated by Tris-glycine SDS-PAGE and stained with GelCode Blue Safe Protein Stain (Part No. 24594). M = MW marker, A = mouse ascites, C = culture supernatant, S = human serum, F = flow-through |
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