Custom Antibody

Q: Does NovoPro guarantee antibody titers?
A: Yes, NovoPro guarantees at least a 1:50,000 antibody titer against peptide sequences that we synthesize/conjugate. Titers are confirmed via ELISA tests that we run.
Q: Do I retain rights to the antisera that I generate?
A: Yes, you will receive all serum / peptides / columns used for generation of your antibody and retain rights to these materials.
Q: Does NovoPro maintain the confidentiality of my project?
A: Yes, NovoPro places great importance in ensuring that our clients' information remains confidential and will never release details of any potential, current or past projects to any outside parties.
Q: Does NovoPro generate Monoclonal antibodies?
A: No, NovoPro focuses in generating custom polyclonal antibodies. Also, our Monospecific antibody production program generates epitope-specific polyclonal antibodies at a fraction of the cost of monoclonal antibody development.
Q: Why are your prices so competitive?
A: NovoPro enjoys economies of scale that allow us to generate high quality antibodies at very competitive prices.
Q: Does NovoPro offer volume discounts?
A: Yes, please inquire with our sales representatives at sales@novoprolabs.com
Q: What is Sodium Azide and should I add it to the antiserum?
A: Sodium Azide is a preservative that prevents bacteria from growing in the serum (bacteria can produce proteases that denature any proteins in the serum, including antibodies). We typically recommend adding azide unless the serum is going to be used directly in cell culture applications. Azide can be removed by using a dialysis membrane with a molecular weight cutoff between 12,000 and 14,000.
Q: Will the titer of antibody increase with additional immunizations?
A: For rabbit projects, the titers remain relatively level after the 1st production bleed and additional immunizations are used to maintain antibody titers rather than to increase them. Specificity does increase between the 1st and 3rd bleeds as part of the affinity maturation process and so antibodies from the third and later bleeds represent the most specific antibodies (which is why the affinity purification is run on the third bleed).
Q: What kind of rabbits does NovoPro use?
A: New Zealand White Rabbits
Q: Will I receive any leftover peptide at the end of the project?
A: Yes, you will receive all leftover peptide at the end of your project, helping ensure the confidentiality of your research. Typically you should receive at least 1 mg of peptide at the end.
Q: How much protein do I need to send?
A: We recommend sending 2-3 mg of the antigen for a rabbit and 5-7 mg for a goat project. We recommend 1 mg/ml as a minimum protein concentration and welcome higher concentrations.
Q: Does the protein need to be soluble?
A: No, insoluble proteins can be used for immunizations.
Q: Should I generate antibodies against a full-length protein or a peptide?
A: Generating antibodies against a full-length protein will provide a pool of antibodies against multiple epitopes from the protein, thereby maximizing the probability of recognizing the endogenous protein in the target assay. However, this larger pool of antibodies does increase the possibility that some of those antibodies will cross-react with other proteins in the assay. Immunizing with a peptide sequence, by contrast, allows the serum to be affinity purified against the peptide sequence, thereby permitting isolation of antibodies that are highly specific to the target protein. The only potential downside to this approach is that there is a risk that the chosen sequence won't correspond to an exposed region of the native protein.
Q: Why is conjugation of the peptide to a carrier protein necessary?
A: The molecular weight of most peptides is too small to generate an immune response in the animal. Conjugation to a carrier protein such as KLH will not only increase the size of the antigen, but increase its immunogenicity as well.
Q: What role does the adjuvant play and what is the difference between CFA and IFA?
A: An adjuvant is a substance that, when combined with the antigen, serves to enhance the immune response against the antigen. Freund's adjuvants are the preferred adjuvant of choice for use in antibody production. Complete Freund's Adjuvant is used only for the first immunization and contains mycobacteria, which stimulates the host's immune system. Incomplete Freund's Adjuvant is used for all subsequent immunizations.
Q: How should I store the antibody / antisera?
A: For short-term storage of the working antibody or of serum that has recently arrived, 2-3 weeks at 4 Celsius degree is recommended. For long-term storage, we recommend storing the serum vials at -20 Celsius degree, which is sufficient for several years. Storage at -80 Celsius degree is fine also, but not necessary except for very long periods of time. The purified antibody should be aliquoted into working quantities and stored according to the guidelines above. The biggest threat to antibody stability is repeated freeze/thaw cycles. To avoid this, we recommend storing ready-to-use aliquots of the antibody and thawing aliquots only as they are needed. We also recommend the use of manual defrost freezers rather than frostless freezers.
Q: Why am I seeing multiple or unexpected bands on a Western with my affinity purified antibody?
A: It isn't uncommon to see multiple bands even when using affinity-purified antibody. This isn't indicative of a problem with the antibody's specificity. Rather, this typically occurs for one of the following reasons: The anti-peptide antibody recognizes a homologous protein in the sample that shares one or more epitopes with the peptide sequence. The native protein is a different molecular weight than previously predicted. The antibody is recognizing either cleaved fragments of the native protein at lower molecular weights or aggregated dimers/trimers of the native protein at higher molecular weights.
Q: Why am I not seeing any bands on my western when assaying with the purified antibody?
A: In cases where no bands are seen (or an antibody doesn't work in a particular assay), these are the three most common explanations: The peptide sequence corresponds to a non-exposed region of the native protein The protein's conformation in the peptide region differs enough that the antibody has trouble recognizing the native protein. The target protein isn't present in the sample.
Q: What is the difference between an IgG purification and an affinity purification?
A: An IgG purification isolates all IgG in the serum, regardless of the specificity. This is advantageous for applications where removing serum proteins will help reduce background noise, but affinity purification against the immunizing protein isn't possible. The affinity purification, by contrast, is advantageous for isolating only those antibodies that recognize a specific epitope. With a peptide, this can yield antibodies that have the same specificity as monoclonal antibodies.

Custom Antibody

Q: Does NovoPro guarantee antibody titers?
A: Yes, NovoPro guarantees at least a 1:50,000 antibody titer against peptide sequences that we synthesize/conjugate. Titers are confirmed via ELISA tests that we run.
Q: Do I retain rights to the antisera that I generate?
A: Yes, you will receive all serum / peptides / columns used for generation of your antibody and retain rights to these materials.
Q: Does NovoPro maintain the confidentiality of my project?
A: Yes, NovoPro places great importance in ensuring that our clients' information remains confidential and will never release details of any potential, current or past projects to any outside parties.
Q: Does NovoPro generate Monoclonal antibodies?
A: No, NovoPro focuses in generating custom polyclonal antibodies. Also, our Monospecific antibody production program generates epitope-specific polyclonal antibodies at a fraction of the cost of monoclonal antibody development.
Q: Why are your prices so competitive?
A: NovoPro enjoys economies of scale that allow us to generate high quality antibodies at very competitive prices.
Q: Does NovoPro offer volume discounts?
A: Yes, please inquire with our sales representatives at sales@novoprolabs.com
Q: What is Sodium Azide and should I add it to the antiserum?
A: Sodium Azide is a preservative that prevents bacteria from growing in the serum (bacteria can produce proteases that denature any proteins in the serum, including antibodies). We typically recommend adding azide unless the serum is going to be used directly in cell culture applications. Azide can be removed by using a dialysis membrane with a molecular weight cutoff between 12,000 and 14,000.
Q: Will the titer of antibody increase with additional immunizations?
A: For rabbit projects, the titers remain relatively level after the 1st production bleed and additional immunizations are used to maintain antibody titers rather than to increase them. Specificity does increase between the 1st and 3rd bleeds as part of the affinity maturation process and so antibodies from the third and later bleeds represent the most specific antibodies (which is why the affinity purification is run on the third bleed).
Q: What kind of rabbits does NovoPro use?
A: New Zealand White Rabbits
Q: Will I receive any leftover peptide at the end of the project?
A: Yes, you will receive all leftover peptide at the end of your project, helping ensure the confidentiality of your research. Typically you should receive at least 1 mg of peptide at the end.
Q: How much protein do I need to send?
A: We recommend sending 2-3 mg of the antigen for a rabbit and 5-7 mg for a goat project. We recommend 1 mg/ml as a minimum protein concentration and welcome higher concentrations.
Q: Does the protein need to be soluble?
A: No, insoluble proteins can be used for immunizations.
Q: Should I generate antibodies against a full-length protein or a peptide?
A: Generating antibodies against a full-length protein will provide a pool of antibodies against multiple epitopes from the protein, thereby maximizing the probability of recognizing the endogenous protein in the target assay. However, this larger pool of antibodies does increase the possibility that some of those antibodies will cross-react with other proteins in the assay. Immunizing with a peptide sequence, by contrast, allows the serum to be affinity purified against the peptide sequence, thereby permitting isolation of antibodies that are highly specific to the target protein. The only potential downside to this approach is that there is a risk that the chosen sequence won't correspond to an exposed region of the native protein.
Q: Why is conjugation of the peptide to a carrier protein necessary?
A: The molecular weight of most peptides is too small to generate an immune response in the animal. Conjugation to a carrier protein such as KLH will not only increase the size of the antigen, but increase its immunogenicity as well.
Q: What role does the adjuvant play and what is the difference between CFA and IFA?
A: An adjuvant is a substance that, when combined with the antigen, serves to enhance the immune response against the antigen. Freund's adjuvants are the preferred adjuvant of choice for use in antibody production. Complete Freund's Adjuvant is used only for the first immunization and contains mycobacteria, which stimulates the host's immune system. Incomplete Freund's Adjuvant is used for all subsequent immunizations.
Q: How should I store the antibody / antisera?
A: For short-term storage of the working antibody or of serum that has recently arrived, 2-3 weeks at 4 Celsius degree is recommended. For long-term storage, we recommend storing the serum vials at -20 Celsius degree, which is sufficient for several years. Storage at -80 Celsius degree is fine also, but not necessary except for very long periods of time. The purified antibody should be aliquoted into working quantities and stored according to the guidelines above. The biggest threat to antibody stability is repeated freeze/thaw cycles. To avoid this, we recommend storing ready-to-use aliquots of the antibody and thawing aliquots only as they are needed. We also recommend the use of manual defrost freezers rather than frostless freezers.
Q: Why am I seeing multiple or unexpected bands on a Western with my affinity purified antibody?
A: It isn't uncommon to see multiple bands even when using affinity-purified antibody. This isn't indicative of a problem with the antibody's specificity. Rather, this typically occurs for one of the following reasons: The anti-peptide antibody recognizes a homologous protein in the sample that shares one or more epitopes with the peptide sequence. The native protein is a different molecular weight than previously predicted. The antibody is recognizing either cleaved fragments of the native protein at lower molecular weights or aggregated dimers/trimers of the native protein at higher molecular weights.
Q: Why am I not seeing any bands on my western when assaying with the purified antibody?
A: In cases where no bands are seen (or an antibody doesn't work in a particular assay), these are the three most common explanations: The peptide sequence corresponds to a non-exposed region of the native protein The protein's conformation in the peptide region differs enough that the antibody has trouble recognizing the native protein. The target protein isn't present in the sample.
Q: What is the difference between an IgG purification and an affinity purification?
A: An IgG purification isolates all IgG in the serum, regardless of the specificity. This is advantageous for applications where removing serum proteins will help reduce background noise, but affinity purification against the immunizing protein isn't possible. The affinity purification, by contrast, is advantageous for isolating only those antibodies that recognize a specific epitope. With a peptide, this can yield antibodies that have the same specificity as monoclonal antibodies.