Overview

Our Custom Phosphospecific Antibody Production package includes everything needed to obtain affinity-purified antibodies that are specific to a phosphorylated epitope. We run an extensive series of ELISA assays on the production bleeds and on the affinity-purified antibodies to ensure that antibodies are specific to the phosphorylated peptide and show no activity against the non-phosphorylated peptide.

Service Items

Service Details

Advantages

Antigen Design
Leading Biology provides antigen design assistance at no additional charge. We use our innovative and proprietary combination of algorithms to analyze your protein sequence and predict which regions are immunogenic, unique and exposed.

Conjugation of Peptide to Carrier Protein
Because peptides are too small to generate an immune response by themselves, conjugation to a carrier protein will be required for immunizations. Leading Biology recommends coupling peptides to KLH (Keyhole Limpet Hemocyanin).

Terminal Cysteine
Leading Biology recommends coupling the phosphor-peptide to the carrier protein via a cysteine that has been added to the N or C-terminal of the peptide. For this reason, we recommend avoiding sequences that contain internal cysteines whenever possible.

Quantity of Peptide
Leading Biology typically synthesizes 40-50 mg of immunograde peptide, with approximately 5-10 mg of each peptide being required for conjugation to the carrier protein, ELISA and affinity purifications. All unused peptides will be delivered to the customer.

Affinity Column
Both the phosphorylated and non-phosphorylated affinity columns are included so that the client can run additional purifications as needed. Longevity of the columns varies, but the best yields are typically obtained within 6-12 months of the project’s completion.

Titer Guarantees
Leading Biology offers one of the highest titer guarantees in the industry, promising at least a 1:50,000 antibody titer against the peptide sequence. For additional details, please read our Guarantee page in our Resources section.

Phospho Purification Strategy
To isolate antibodies specific to the phospho epitope, the serum will first be affinity purified against the non-phosphorylated peptide prior to being run through the phosphorylated peptide column. Comprehensive ELISA testing against both peptides confirms the specificity of the purified antibodies.

Purified Antibody Yields
On average, 2-3 mg of peptide-specific antibody will be isolated from affinity purification of 25 ml of antiserum. However, because antibodies specific to the phosphosite will represent only a portion of antibodies against the phosphopeptide sequence, the yield of phosphospecific antibodies will typically be around 0.5 – 1 mg.

General Purification Strategy
Unless one animal has particularly high antibody titers against the peptide, we recommend affinity-purifying a pool of 25 ml serum taken in a 1:1 ratio from both animals to ensure maximum epitope variability.

Why leading biology?
With over 5 years of experience in custom antibodies, we offer numerous benefits for developing your custom antibody:
• Our industry leading titer guarantees of 1:50,000 eliminate the risk of not obtaining antibodies against peptide antigens.

• Our innovative antigen prediction algorithms maximize the probability of targeting peptide sequences that correspond to exposed regions of native proteins.
• Our economies of scale allow us to pass cost savings to you and help maximize your budget.
• 100% of services are transparent throughout.
• Antibodies from our facility have been cited in many published research papers.
• By outsourcing antibody production and characterization needs to us, you can spend more time focused on your research.

• Our ideal location in California ensures that animals benefit from outdoor facilities and mild year-round temperatures.

• Our Monospecific antibody production program generates epitope-specific polyclonal antibodies with superior affinity to monoclonal antibodies and at a fraction of the cost.

• Confidentiality is a top priority for all projects. Antibodies that we manufacture belong to you and will not be commercialized.

Contact Information

Please obtain a quote before ordering, and refer to the quote number when you place an order.
Orders are typically confirmed within 12 hours.
Have a Question? Email us info@leadingbiology.com
Order Products: Order Related Products
By Phone: 1-661-524(LBI)-0262 (USA)