Mab Anti Tag

The Mab Anti Tag Coated Surfaces are useful for screening sera for antibodies to the fusion protein and are ideal to determine the presence and concentration of tag-fusion proteins from cell lysates.

The development of recombinant DNA technology has permitted the production of proteins that were originally extracted from tissues and secretions to be produced synthetically with high purity and yield. Multiple-host expression systems including mammalian, insect, bacterial, yeast, algal, and cell-free systems are available and are easy to transform with a DNA vector containing the gene of interest. Typically, an affinity tag sequence (additional amino acids, a functional domain, or a whole protein) is cloned in frame with the DNA sequence of the target protein, and will flank either the N- or C-terminus, to aid in purification and analysis. These additions are known as fusion tags.

The properties of fusion tags allow tagged proteins to be manipulated easily in the laboratory. The availability of antibodies to fusion tags and their easy immobilization on polystyrene surface has permitted their use in downstream detection and assay methods, eliminating the need to obtain or develop a probe for each specific recombinant protein.

Coated Surface for Mab Anti Tag Proteins are an ideal tool to carry out ELISA assays to determine the presence and concentration of tag-fusion proteins from cell lysates. Also, using these plates to immobilize tag fusion proteins, is useful for screening sera for antibodies to the fusion protein.

This is the available configuration of Mab Anti Tag 96-Well Coated Plate products by Biomat.

Click on the configuration link to scroll down the page to the relative section.

• Anti Tag Coated 96-Well Plates
• Mab αGST Coated 96-Well Plates
• Mab αHis Tag Coated 96-Well Plates
• Mab αDYKDDDDK Coated 96-Well Plates

Other 96-Well Plates

Biomat’s Mab Anti Tag Coated 96-Well Plates have the following characteristics:

• Ready to use
• Manufactured under ISO:9001 guidelines
• Post-coated (blocked) for low non specific binding and long-term stability
• All lots are tested for uniformity and reproducibility
• Certificate of Quality is released for every lot
• For Research Use Only

Also, Coated Surface for Mab Anti Tag Test Products ensures:

Storage and Stability

The microplates, if unopened, are stable at 2-8°C until the expiration date printed on the label. If opened, store in closed pouch with desiccant and use within the expiration date.

Click on the below  link to scroll down the page to the relative section.

Mab αGST (Glutathione S-Transferase) 96-Well Plates

Mab αHis (Polyhistidine) 96-Well Plates

Mab αDYKDDDDK 96-Well Plates

GST is a 211 amino acid protein (M.W. 26 kDa) whose DNA sequence is frequently integrated into expression vectors for production of recombinant proteins. The result of expression from this vector is a GST-tagged fusion protein in which the functional GST protein is fused to the N-terminus of the recombinant protein.

Mab αGST (Glutathione S-Transferase) 96-Well Plates are designed to specifically bind GST tagged proteins and GST. 

Examples of applications for Mab αGST Coated 96-Well Plates:

• ideal to bind proteins with GST (Glutathione S-Transferase) tag
• pre-purification of cell lysates is not necessary before screening and analysis of recombinant GST-tagged protein expression by ELISA using the plates
• after immobilizing GST fusion proteins, the plates are useful for screening for sera antibodies to the fusion protein

General ELISA procedures using anti-GST tag coated plates

Coating

Mouse monoclonal anti-GST is coated using 100 µl/well.

Binding capacity

Microplate was saturated with GST at a concentration of 1.0 µg/ml (100 ng/well) in an ELISA format using goat anti GST-HRP as detector and TMB as substrate.

Sensitivity

GST was detected at a concentration significantly above background in an ELISA format using goat anti GST-HRP as detector and TMB as substrate.

see binding capacity and sensitivity technical note

Uniformity

Microplates show a CV% less than 7 when used as a sandwich of GST in an ELISA format using goat anti GST-HRP as detector and TMB as substrate.

Reagent Compatibility

Some reagents may interfere with the test results. Check the reagents concentration in sample according to the reagent compatibility tests table. Dialyse or dilute samples if needed.

see reagent compatibility table

The His-tag is an amino acid motif in proteins and is one of the simplest and most widely used purification tags. Typically consist of four, five, six or more consecutive histidine residues (hexa-His (6 His), penta-His (5 His) and tetra-His (4 His) respectively) and  often is present at the N- or C-terminus of the protein.

Mab αHis (polyhistidine) 96-Well Plates are designed to specifically bind Histidine tagged proteins with His tag located at N-terminus or C-terminus. 

Examples of applications for Mab αHis Coated 96-Well Plates:

• ideal to bind proteins with HIS (Poly histidine) tag
• pre-purification of cell lysates is not necessary before screening and analysis of recombinant HIS-tagged protein expression by ELISA using the plates
• after immobilizing HIS fusion proteins, the plates are useful for screening for sera antibodies to the fusion protein
• it can be applied for analysis of different length of His tag including: hexa-His (6 His), penta-His (5 His) and tetra-His (4 His)

General ELISA procedures using anti-HIS tag coated plates

Coating

Mouse monoclonal anti-HIS is coated using 100 µl/well.

Binding capacity

Microplate was saturated with HSA-His Tag to a concentration between 2.0 and 4.0 µg/ml (200 – 400 ng/well) in an ELISA format using rabbit anti HSA-HRP as detector and TMB as substrate.

Sensitivity

HSA-His tag was detected at a concentration significantly above background in an ELISA format using rabbit anti HSA-HRP as detector and TMB as substrate.

see binding capacity and sensitivity technical note

Uniformity

Microplates show a CV% less than 6 when used as a sandwich of HSA-His Tag in an ELISA format using rabbit anti HSA-HRP as detector and TMB as substrate.

Reagent Compatibility

Some reagents may interfere with the test results. Check the reagents concentration in sample according to the reagent compatibility tests table. Dialyse or dilute samples if needed.

see reagent compatibility table

The DYKDDDDK tag (also called octapeptide tag or FLAG-tag) is a short and hydrophilic epitope tag that consists of eight amino acids (Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys).

The DYKDDDDK tag is one of the most specific tags that can be added to a protein using recombinant DNA Technology.

Mab αDYKDDDDK 96-Well Plates designed to specifically bind DYKDDDDK tagged proteins with DYKDDDDK tag located at N-terminus or C-terminus. 

Examples of applications for Mab αDYKDDDDK Coated 96-Well Plates:

• ideal to bind proteins with DYKDDDDK tag
• pre-purification of cell lysates is not necessary before screening and analysis of recombinant DYKDDDDK tagged protein expression by ELISA using the plates
• after immobilizing DYKDDDDK fusion proteins, the plates are useful for screening for sera antibodies to the fusion protein

General ELISA procedures using anti-DYKDDDDK coated plates

Coating

Mouse monoclonal anti-DYKDDDDK is coated using 100 µl/well.

Binding capacity

Microplate was saturated with Recombinant Human Flag Ubiquitin at a concentration close to 6.0 µg/ml (600 ng/well) in an ELISA format using a Mab anti Ubiquitin-Biotin plus Streptavidin-HRP as detector and TMB as substrate.

Sensitivity

Recombinant Human Flag Ubiquitin was detected at a concentration significantly above background in an ELISA format using a Mab anti Ubiquitin-Biotin plus Streptavidin-HRP as detector and TMB as substrate.

see binding capacity and sensitivity technical note

Uniformity

Microplates show a CV% less than 5 when used as a sandwich of Recombinant Human Flag Ubiquitin in an ELISA format using a Mab anti Ubiquitin-Biotin plus Streptavidin-HRP as detector and TMB as substrate.

Reagent Compatibility

Some reagents may interfere with the test results. Check the reagents concentration in sample according to the reagent compatibility tests table. Dialyse or dilute samples if needed.

see reagent compatibility table