Documents & Support

Preparation of a verifiable peptide-protein immunogen: direction-controlled conjugation of a synthetic fragment of the monitor peptide with myoglobin and application for sequence analysis. Citations & References

  • Authors: Iwai K, Fukuoka S, Fushiki T, Kido K, Sengoku Y, Semba T
  • Journal: Anal Biochem
  • PubMed ID: 3407924
Catalog # S1533

Preparation of a verifiable peptide-protein immunogen: direction-controlled conjugation of a synthetic fragment of the monitor peptide with myoglobin and application for sequence analysis Citations & References

  • Authors: Iwai, K., Fukuoka, S.-I., Fushiki, T., Kido, K., Sengoku, Y. and Semba, T.
  • Journal: Anal Biochem
  • PubMed ID: 0
Catalog # 22317

Toll-Like Receptor 7/8 (TLR7/8) and TLR9 Agonists Cooperate To Enhance HIV-1 Envelope Antibody Responses in Rhesus Macaques. Citations & References

  • Authors: Moody MA, Santra S, Vandergrift NA, Sutherland LL, Gurley TC, Drinker MS, Allen AA, Xia SM, Meyerhoff RR, Parks R, Lloyd KE, Easterhoff D, Alam SM, Liao HX, Ward BM, Ferrari G, Montefiori DC, Tomaras GD, Seder RA, Letvin NL, Haynes BF,
  • Journal: J Virol
  • PubMed ID: 24390332
Catalog # LPC0005M

General Protein Expression Experiment Protocol

Recombinant respiratory syncytial viruses lacking the C-terminal third of the attachment (G) protein are immunogenic and attenuated in vivo and in vitro. Citations & References

  • Authors: Elliott MB; Pryharski KS; Yu Q; Parks CL; Laughlin TS; Gupta CK; Lerch RA; Randolph VB; LaPierre NA; Dack KM; Hancock GE
  • Journal: Journal of Virology
Catalog #
  • 4339386
  • 4413750
  • N8050400(Discontinued)
  • N8050200(Discontinued)
  • N8050002(Discontinued)
  • ZGEXSC9700384W
  • ZGEXSC9700384W3Y
  • N8050002R
  • 4441166

How are peptide/phosphopeptide antibody competition studies performed? Product FAQ

Answer

Invitrogen Phosphorylation Site-Specific Antibodies detect the phosphorylation of a particular amino acid residue within a particular protein. The immunogen used to raise a Phosphorylation Site Specific Antibody is a synthetic peptide, the sequence of which corresponds to the phosphorylated residue plus the surrounding residues of the protein of interest.  Because the immunogen corresponds to the phosphorylated amino acid residue within the context of the amino acid sequence of the protein, the antibody produced in response to the immunogen is specific to the phosphorylated protein, rather than to a phosphorylated amino acid residue.

Phosphorylation Site Specific Antibodies are useful for the detection of protein phosphorylation by Western blotting and other antibody based procedures, including immunohistochemistry, flow cytometry, and ELISA. The specificity of a Phosphorylation Site Specific Antibody for a phosphorylated protein can be verified by performing a simple peptide competition experiment. In this peptide competition experiment, the antibody of interest is pre-incubated with either (1) water (total signal control), (2) the phosphopeptide immunogen, or (3) the corresponding non-phosphopeptide.  The pre-incubated antibody is then used as the primary antibody in Western blotting or other antibody based procedure.  If the specificity of the antibody and the sample conditions are optimal, the phosphopeptide immunogen will be observed to block binding of the antibody to the phosphorylated protein, while the non-phosphopeptide will be observed to have no effect.

In most peptide competition experiments with Invitrogen control peptides, the phosphopeptide and non-phosphopeptide are used at a 200 fold molar excess over the antibody.  Please note that while a 200 fold molar excess of peptide to antibody is found to be satisfactory for most applications, use of a 500 fold molar excess of peptide to antibody may be optimal for a particular system. It is also important to note that with this and any antibody based procedure, some optimization of antibody concentration and incubation times may be necessary. Calculation for the determination of the 200 fold molar excess of peptide to antibody usually involves the following assumptions: The molecular mass of an IgG molecule is 150,000 daltons. Each mole of antibody binds two moles of peptide. The Phosphorylation Site Specific Antibody is used at a concentration of 0.5 mg/mL. The optimal antibody concentration for use in peptide competition experiments is a concentration which is below saturating as determined by previous experiments in your system.  If an optimal concentration has not been determined, it is suggested that the concentration provided on the antibody Product Analysis Sheet be used. 

Answer Id:: E5122

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Can you provide the exact immunogen sequence for IL-2 Polyclonal Antibody (Cat. No. PA5-85127)? Product FAQ

Answer

The exact immunogen sequence for IL-2 Polyclonal Antibody (Cat. No. PA5-85127) is proprietary. However, we can divulge that the immunogen is a recombinant protein encompassing a sequence within the center region of human IL-2.

Answer Id:: E19268

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Cationization of protein antigens. I. Alteration of immunogenic properties. Citations & References

  • Authors: Muckerheide, et al.
  • Journal: J Immunol
  • PubMed ID: 0
Catalog # 77150

Can you please let me know whether the immunogen sequence of the ANGPTL4 Polyclonal Antibody (Cat. No. 40-9800) is before or after the 168 amino acid of the protein? Product FAQ

Answer

The immunogen used was from amino acids 259-274 of the protein sequence in Uniprot ID Q9BY76.

Answer Id:: E19211

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Identification and expression of immunogenic proteins of a disease-associated marine turtle herpesvirus. Citations & References

  • Authors: Coberley SS, Condit RC, Herbst LH, Klein PA,
  • Journal: J Virol
  • PubMed ID: 12239336

Some of the Gibco Recombinant Proteins are not biologically active. Can these be used for cell culture? Product FAQ

Answer

No, cell culture usage requires biologically active protein. These proteins are for use as a protein standard or immunogen.

Answer Id:: E5572

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Independent loss of immunogenic proteins in Mycobacterium ulcerans suggests immune evasion Citations & References

  • Authors: Huber, CA; Ruf, MT; Pluschke, G; Kaeser, M
  • Journal: Clinical and Vaccine Immunology
Catalog #
  • 4339386
  • 4413750
  • N8050400(Discontinued)
  • N8050200(Discontinued)
  • N8050002(Discontinued)
  • ZGEXSC9700384W
  • ZGEXSC9700384W3Y
  • N8050002R
  • 4441166

Identification, characterization, and immunogenicity of the lactoferrin-binding protein from Helicobacter pylori. Citations & References

  • Authors: Dhaenens L, Szczebara F, Husson MO
  • Journal: Infect Immun
  • PubMed ID: 9009306
Catalog #

Can I obtain the peptide sequence of the immunogen? Product FAQ

Answer

We normally do not disclose the amino acid (aa) sequences of our immunogens, nor do we provide any other specific information about them. The immunogen-related information that we do provide is listed in the manual for the antibody. We have chosen not to reveal the peptide sequences for most of our antibody immunogens, primarily because we do not seek patent protection for them. However, Technical Support can provide an approximate location of the immunogen in the protein (e.g., between aa 23 and aa 100), the general location of the epitope (e.g., C- or N-terminus, cytoplasmic or membrane domain), and other related information, if available.

Answer Id:: E12586

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Sequence and immunogenicity of the 70-kDa heat shock protein of Mycobacterium leprae. Citations & References

  • Authors: McKenzie KR; Adams E; Britton WJ; Garsia RJ; Basten A
  • Journal: Journal of Immunology (Baltimore, Md. : 1950)
Catalog #
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