Peptide Purity
Peptide Purity
Phospho-specific antibody production against a peptide containing a phospho-residue is a bit more complicated than traditional peptide antibody production. The animals can produce antibodies against additional epitopes in the peptide sequence, as well as produce antibodies that do not contain the modified phospho-residue. While a pool of phospho and non-phospho antibodies can be difficult to work with, Bio-Synthesis only offer highly purified phosphor-peptide for this factor, and if purified from the anti-serum the non-phospho antibody can be very helpful for studying protein pathways.
Factors to consider:
Length: Use a peptide with a length of 12 residues or less. Length is important due to the number of epitopes available to the host animal's immune system. The minimum length of an epitope is 6-8 amino acids; using a sequence with 12 or fewer residues ensures that the phosphorylated residue will be included in all antibodies produced.
Position of Phosphorylated Residue: Place the phosphorylated residue in the middle of the peptide sequence. This increases the probability that all antibodies produced against the sequence will have epitopes including the phosphorylated residue.
Peptide Purity: Use a phospho peptide of >90% purity and a non-phosphopeptide can be >85%; We highly recommend our client use our superclass phospho-specific peptide package to ensure that the antibodies are produced against the exact sequence of interest. This is also true with any peptide modifications which require two columns affinity specific purification (e.g. acetylated lysine)
Antibody Purification: Serum containing a mixture of phospho and non-phospho antibodies in various proportion, corresponding to the non-phosphorylated version ( formed in the animal via native phosphatase activity of the host) and also pan-specific antibodies, which can be formed against the rest of the peptide, that do not contain the phosphorylated residue. In order to obtain to purify the desired antibody generated against the phosphorylated peptide, two step purification process is been used in our laboratory which is designed to detect the presence of modified amino acid in a sequence specific context.
Step 1 Phospho peptide column: Bind the phospho peptide to gel matrix for column. This will bind only the phospho-specific antibodies and the pan-specific antibodies. The non-phosphospecific antibodies will flow-through through the phospho column. The Phospho and pan specific antibodies can be eluted from the column together with the entire complement of additional IgGs that were isolated from the rabbit serum.
Step 2 Non-phospho peptide column: Bind the non-phospho peptide to gel matrix for purification. This column will bind the pan specific antibody only and the phospho specific antibodies that do not bind to the column will be in the flow-through which contains our desired phospho-specific antibodies.
Antibody Characterization The final stage of custom phospho-specific antibody production is the characterization of the antibodies. We use two methods:
%u2022 ELISA: Phospho peptide to run an ELISA on the serum, phospho affinity purified antibody, and the flow-through resulting from the phospho column. This process is included in our custom phospho-specific antibody packages: FIIP-PH-RA120I, SIIP-PH-RA120I
%u2022 Western Blot: The first Western blot analysis simply demonstrates that there are no cross-reacting bands labeled by the antibody in a homogenate or cell lysate. The second level of specificity uses Western blots of phospho- and dephospho-proteins to demonstrate the selectivity of the phospho-specific antibody. This process included in our custom phospho-specific antibody packages: SIIW-PH-RA120I
All these packages isolate both phospho-specific antibodies and the non-phospho antibodies for your protein. The corresponding antibodies can be used separately or in conjunction to study your protein's regulatory pathways.
Phospho-specific antibody production against a peptide containing a phospho-residue is a bit more complicated than traditional peptide antibody production. The animals can produce antibodies against additional epitopes in the peptide sequence, as well as produce antibodies that do not contain the modified phospho-residue. While a pool of phospho and non-phospho antibodies can be difficult to work with, Bio-Synthesis only offer highly purified phosphor-peptide for this factor, and if purified from the anti-serum the non-phospho antibody can be very helpful for studying protein pathways.
Factors to consider:
Length: Use a peptide with a length of 12 residues or less. Length is important due to the number of epitopes available to the host animal's immune system. The minimum length of an epitope is 6-8 amino acids; using a sequence with 12 or fewer residues ensures that the phosphorylated residue will be included in all antibodies produced.
Position of Phosphorylated Residue: Place the phosphorylated residue in the middle of the peptide sequence. This increases the probability that all antibodies produced against the sequence will have epitopes including the phosphorylated residue.
Peptide Purity: Use a phospho peptide of >90% purity and a non-phosphopeptide can be >85%; We highly recommend our client use our superclass phospho-specific peptide package to ensure that the antibodies are produced against the exact sequence of interest. This is also true with any peptide modifications which require two columns affinity specific purification (e.g. acetylated lysine)
Antibody Purification: Serum containing a mixture of phospho and non-phospho antibodies in various proportion, corresponding to the non-phosphorylated version ( formed in the animal via native phosphatase activity of the host) and also pan-specific antibodies, which can be formed against the rest of the peptide, that do not contain the phosphorylated residue. In order to obtain to purify the desired antibody generated against the phosphorylated peptide, two step purification process is been used in our laboratory which is designed to detect the presence of modified amino acid in a sequence specific context.
Step 1 Phospho peptide column: Bind the phospho peptide to gel matrix for column. This will bind only the phospho-specific antibodies and the pan-specific antibodies. The non-phosphospecific antibodies will flow-through through the phospho column. The Phospho and pan specific antibodies can be eluted from the column together with the entire complement of additional IgGs that were isolated from the rabbit serum.
Step 2 Non-phospho peptide column: Bind the non-phospho peptide to gel matrix for purification. This column will bind the pan specific antibody only and the phospho specific antibodies that do not bind to the column will be in the flow-through which contains our desired phospho-specific antibodies.
Antibody Characterization The final stage of custom phospho-specific antibody production is the characterization of the antibodies. We use two methods:
%u2022 ELISA: Phospho peptide to run an ELISA on the serum, phospho affinity purified antibody, and the flow-through resulting from the phospho column. This process is included in our custom phospho-specific antibody packages: FIIP-PH-RA120I, SIIP-PH-RA120I
%u2022 Western Blot: The first Western blot analysis simply demonstrates that there are no cross-reacting bands labeled by the antibody in a homogenate or cell lysate. The second level of specificity uses Western blots of phospho- and dephospho-proteins to demonstrate the selectivity of the phospho-specific antibody. This process included in our custom phospho-specific antibody packages: SIIW-PH-RA120I
All these packages isolate both phospho-specific antibodies and the non-phospho antibodies for your protein. The corresponding antibodies can be used separately or in conjunction to study your protein's regulatory pathways.
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About BSI
"The use of state-of-the art technology has allowed Bio-Synthesis, Inc. to rapidly gain recognition for its manufacturing quality & dependability of delivery."
Founded in 1984, Bio-Synthesis, Inc. was first known as OCS Laboratories. It was the first producer of commercially available synthetic DNA and, in 1985, became a producer of synthetic peptides. In 1989, OCS incorporated as Bio-Synthesis, Inc. and moved its laboratories to Lewisville, Texas. Today, BSI occupies 10,000 square feet of modern laboratory space (completed in 1995) located 20 miles from downtown Dallas and 10 miles from D/FW airport. Among the surrounding academic institutions are: University of Texas Southwestern Medical School (12 miles), University of North Texas (10 miles), and University of North Texas Health Science Center (25 miles). These universities provide excellent library resources. Since its inception, BSI has steadily expanded its product lines and services to meet the growing needs of the molecular biology community. It has maintained its position as an aggressive, innovative company in a highly competitive marketplace without sacrificing quality. In the beginning, our primary emphasis was on synthesizing high quality DNA primers and linkers, which were the initial uses of oligos. Today, newer technologies, such as gene construction, PCR, mutagenesis, combinatorial libraries, dye/adduct labeling, DNA microarrays, peptide-nucleic acid chimeras, etc. have challenged the molecular biology field. In response, BSI has branched into several related areas including DNA paternity testing, DNA HLA typing, PNA synthesis, genomic sequencing, fluorescence-based genotyping, custom organic synthesis and other molecular biology based applications.
The Research and Development Division comprises a number of Ph.D. and M.D. professionals with backgrounds in molecular biology, immunology, nucleic acids, peptide synthesis, general organic chemistry, and automated amino acid sequencing. The Molecular Biology Division is set up to handle most modern experimental procedures; the Immunology Division specializes in immunodiagnostic-based reagents.
Not only has BSI continued to provide quality DNA products and services for the research community, but it has also become a world leader in providing custom peptide products and services. Using state-of-the-art solid phase peptide chemistries, BSI provides high quality custom peptides, performs carrier conjugations, antipeptide antibody production, antigenic peptide design, synthesis of long and/or modified peptides, MALDI TOF analysis, contract research, consultation services and more.
Biomedical researchers worldwide in universities, biotech companies, private clinics, and government agencies use products from Bio-Synthesis, Inc. in studies ranging from PCR diagnostics to cancer research and the Human Genome Project. Contact us to discover how BSI can customize products/services to suit your research requirements.
For more detail please visit our About Us
Please Contact Us :
Address: 612 E. Main street
City: Lewisville
State: Texas
Country: USA
Postal Code: 75057
Phone Number: 972-420-8505
Founded in 1984, Bio-Synthesis, Inc. was first known as OCS Laboratories. It was the first producer of commercially available synthetic DNA and, in 1985, became a producer of synthetic peptides. In 1989, OCS incorporated as Bio-Synthesis, Inc. and moved its laboratories to Lewisville, Texas. Today, BSI occupies 10,000 square feet of modern laboratory space (completed in 1995) located 20 miles from downtown Dallas and 10 miles from D/FW airport. Among the surrounding academic institutions are: University of Texas Southwestern Medical School (12 miles), University of North Texas (10 miles), and University of North Texas Health Science Center (25 miles). These universities provide excellent library resources. Since its inception, BSI has steadily expanded its product lines and services to meet the growing needs of the molecular biology community. It has maintained its position as an aggressive, innovative company in a highly competitive marketplace without sacrificing quality. In the beginning, our primary emphasis was on synthesizing high quality DNA primers and linkers, which were the initial uses of oligos. Today, newer technologies, such as gene construction, PCR, mutagenesis, combinatorial libraries, dye/adduct labeling, DNA microarrays, peptide-nucleic acid chimeras, etc. have challenged the molecular biology field. In response, BSI has branched into several related areas including DNA paternity testing, DNA HLA typing, PNA synthesis, genomic sequencing, fluorescence-based genotyping, custom organic synthesis and other molecular biology based applications.
The Research and Development Division comprises a number of Ph.D. and M.D. professionals with backgrounds in molecular biology, immunology, nucleic acids, peptide synthesis, general organic chemistry, and automated amino acid sequencing. The Molecular Biology Division is set up to handle most modern experimental procedures; the Immunology Division specializes in immunodiagnostic-based reagents.
Not only has BSI continued to provide quality DNA products and services for the research community, but it has also become a world leader in providing custom peptide products and services. Using state-of-the-art solid phase peptide chemistries, BSI provides high quality custom peptides, performs carrier conjugations, antipeptide antibody production, antigenic peptide design, synthesis of long and/or modified peptides, MALDI TOF analysis, contract research, consultation services and more.
Biomedical researchers worldwide in universities, biotech companies, private clinics, and government agencies use products from Bio-Synthesis, Inc. in studies ranging from PCR diagnostics to cancer research and the Human Genome Project. Contact us to discover how BSI can customize products/services to suit your research requirements.
For more detail please visit our About Us
Please Contact Us :
Address: 612 E. Main street
City: Lewisville
State: Texas
Country: USA
Postal Code: 75057
Phone Number: 972-420-8505
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Custom Antibody
As a leading supplier of genomic and proteomic research products, BSI is able to offer its customers integrated solutions for custom DNA synthesis, custom RNA synthesis, custom polyclonal antibody production, custom peptide synthesis, molecular diagnostic detection kits, DNA Real-Time PCR products, DNA microarray services and a wide range of custom DNA-based diagnostic kits and consumables.
The proteomic division is experienced in the design of peptides, both for antibody production and bioactive peptide synthesis, e.g cosmetic peptides. While the custom peptide synthesis division has the ability to produce thousands of modified or non-modified peptides per day for research applications, BSI also provides diagnostic/therapeutic GMP custom peptide production. Synthesis of custom peptides results in quantities ranging from milligrams to multi-kilograms. The staff in the custom antibody division provide expert support in the selection of peptide antigens for target antibody production and offer convenient custom-bundled packages with a wide range of host animals. In addition, peptide, antibody and protein arrays are also amongst its expertise.
BSI is a U.S. based company whose primary emphasis is on the synthesis of high quality custom products for a variety of applications and a full-service contract manufacturing organization (CMO) delivering research, development and production services for cGMP clinical batch manufacturing.
BIO-SYNTHESIS
BIO-SYNTHESIS, INC., is a leading life science products company with over 20 years of experience in the design and synthesis of Custom Peptide, small molecules and reagents for small scale research and bulk pharmaceutical trials. Using state of the art technology in our well-equipped laboratories.
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BIO-SYNTHESIS
BIO-SYNTHESIS, INC. is a leading life science products company with over 20 years of experience in the design and synthesis of Custom Peptide, small molecules and reagents for small scale research and bulk pharmaceutical trials. Using state of the art technology in our well-equipped laboratories.
Please visit our Blog on Blogspot Bio-Synthesis
Please visit our Blog on Blogspot Bio-Synthesis
