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Imject Maleimide-Activated Mariculture KLH and Kit


SMCC-activated KLH carrier protein for immunogen prep with cysteine peptides.

Maleimide-activated KLH carrier protein for immunogen preparation with sulfhydryl peptides

Thermo Scientific Imject Maleimide-Activated mcKLH products enable conjugation of sulfhydryl-containing peptide haptens to the highly immunogenic keyhole limpet hemocyanin (KLH) carrier protein for immunization and antibody production.

The high-quality mariculture KLH is rendered sulfhydryl-reactive by chemical modification with Sulfo-SMCC crosslinker, and the resulting maleimide-activated protein is then stabilized and lyophilized in phosphate buffer. Maleimide groups form covalent crosslinks with sulfhydryl (-SH) moieties on cysteine residues of peptides and other thiol-containing haptens. Numerous antigen molecules can be conjugated to each KLH molecule to make them highly immunogenic and increase the likelihood of eliciting immunized animals to produce high-titer antisera to the epitopes of interest. Select from among several package sizes of activated KLH protein, or choose the complete conjugation kit that includes optimized buffers and trouble-free spin desalting columns.

Highlights:

  • Convenient kit – containing stabilized, lyophilized Maleimide-Activated mcKLH (in PBS with stabilizer) and accessory components to easily prepare ready-to-use immunogens for immunization
  • Optimized activation – specifications require that each lot is activated with more than 400 moles of sulhydryl-reactive maleimide per mole of KLH, ensuring consistently high levels of conjugation
  • Sulfhydryl-reactive – engineered for conjugating cysteine-containing peptide antigens to make effective immunogens for high-titer antibody production
  • Simple, rapid procedure – complete the hapten-carrier conjugation reaction in two hours to yield sufficient immunogen for several injections and booster immunizations of host animals
  • High-quality carrier protein – KLH is isolated from the mollusk Megathura crenulata (keyhole limpet); mcKLH is a high-quality form of KLH that is harvested from limpets grown in mariculture
  • Highly immunogenic – KLH has a high molecular mass (4.5 x 105 to 1.3 x 107 Daltons; aggregates of 350 and 390 kDa subunits) and elicits a stronger immune response than BSA or ovalbumin
  • Excellent solubility – our preparations of mariculture KLH are more soluble than typical forms and we also offer a PEGylated form for even greater hydrophilicity and solubility

Product Details:

Carrier proteins are large, complex molecules capable of stimulating an immune response upon injection. Successful production of antibodies specific to small antigens (i.e., peptides or drug compounds) requires that these haptens be covalently conjugated to a larger, more complex molecule (usually a protein) to make them immunogenic. Carrier proteins are chosen based on immunogenicity, solubility, and whether an adequate level of hapten-carrier conjugation can be achieved.

Keyhole limpet hemocyanin (KLH) is widely used as a carrier protein for conjugation to haptens and other antigens to make them more immunogenic for the purpose of antibody production. Because of its large mass and complexity, KLH elicits a stronger immune response than other carrier proteins. Because the protein is derived from a mollusk, it is phylogenetically distant from mammalian species and less likely to produce antibodies that will cross-react with typical target samples in assays.

Structure and conjugation reaction scheme of Thermo Scientific Imject Maleimide-Activated mcKLH. High-quality mariculture KLH is modified with Sulfo-SMCC crosslinker to produce the maleimide-activated carrier protein, which is supplied lyophilized. Addition of peptides or other molecules containing reduced sulfhydryl groups results in efficient and specific reaction to form stable thioether bonds. Each carrier protein molecule contains several hundred maleimide activations, allowing conjugation of many peptide hapten molecules per KLH molecule. Maleimide-Activated KLH is prepared using Sulfo-SMCC, making it reactive to peptides with terminal cysteines.

 

Structure of Thermo Scientific Imject PEGylated Maleimide-Activated mcKLH. Inclusion of polyethylene glycol (PEG) in the spacer arm of added maleimide groups increases the solubility and hydrophilicity of the carrier protein. Learn more about polyethylene glycol and PEGylation. Structure of PEGylated Maleimide-Activated mcKLH.

 

More Product Data

References:

  1. Harlow, E. and Lane, D. (1988). Antibodies: A Laboratory Manual. Cold Spring Harbor, New York: Cold Spring Harbor Laboratory. Chapter 5 discusses the use of carrier proteins.
  2. Hermanson, G.T. (2008). Bioconjugate Techniques. 2nd edition, Academic Press, New York. (Part No. 20036). Chapter 19 discusses carrier protein uses and the maleimide-activation chemistry.
  3. Lagace, T.A., et al. (2006). Secreted PCSK9 decreases the number of LDL receptors in hepatocytes and inlivers of parabiotic mice. J. Clin. Invest. 116:2995-3005.
  4. Casais, R., et al. (2005). Gene 5 of the avian coronavirus infectious bronchitis virus is not essential for replication. J. Virol. 79:8065-8078.
  5. Becker, E.B.E., et al. (2004). Characterization of the c-Jun N-terminal kinase-BimEL signaling pathway in neuronal apoptosis. J. Neurosci. 24:8762-8770.
  6. Chang, S.C., et al. (1994). Cartilage-derived morphogenetic proteins. J. Biol. Chem. 45:28227-28234.
  7. Heymach, Jr., J.V., et al. (1995). The biosynthesis of neurotrophin heterodimers by transfected mammalian cells. J. Biol. Chem. 270:12297-12304.
  8. Joshi, H.C., et al. (1992). g-Tubulin is a centrosomal protein required for cell cycle-dependent microtubule nucleation. Nature 356:80-83.
  9. Juge-Aubry, C.E., et al. (1995). Peroxisome proliferator-activated receptor mediates cross-talk with thyroid hormone receptor by competition for retinoid X receptor. J. Biol. Chem. 270:18117-18122.
  10. Karim, A.S. (1995). Maleimide-mediated protein conjugates of a nucleoside triphosphate gamma-S and an internucleotide phosphorothioate diester. Nucl. Acid. Res. 23:2037-2040.
  11. Slunt, H.H., et al. (1994). Expression of a ubiquitous, cross-reactive homologue of the mouse ß-amyloid precursor protein (APP). J. Biol. Chem. 269:2637-2644.
  12. Wojcikiewicz, R.J.H. (1995). Type I, II, and III Inositol 1, 4, 5-triphosphate receptors are unequally susceptible to down-regulation and are expressed in markedly different proportions in different cell types. J. Biol. Chem. 270:11678-11683.
  13. Chu, H., et al. (2006). Gamma-Parvin is dispensable for hematopoiesis, leukocyte trafficking, and T-cell-dependent antibody response. Mol. Cell. Biol. 26:1817-1825. [+SulfoLink]
  14. Lampe, P.D., et al. (2006). Analysis of Connexin43 phosphorylated at S325, S328 and S330 in normoxic and ischemic heart. J. Cell Sci. 119:3435-3442. [+SulfoLink]
  15. Nakahara, K., et al. (2006). Maternal ghrelin plays an important role in rat fetal development during pregnancy. Endocrinology 147:1333-1342.
  16. Das, S., et al. (2006). Infection-dependent nuclear localization of US17, a member of the US12 family of human cytomegalovirus-encoded seven-transmembrane proteins. J. Virol. 80:1191-1203.
  17. Li, X-H, et al. (2004). Estrogen directly induces expression of retinoic acid biosynthetic enzymes, compartmentalized between the epithelium and underlying stromal cells in rat uterus. Endocrinology 145:4756-4762.
  18. Schofield, L., et al. (2002). Synthetic GPI as a candidate anti-toxic vaccine in a model of malaria. Nature 418:785-789.
  19. Quill, T.A., et al. (2001). A voltage-gated ion channel expressed specifically in spermatozoa. Proc. Natl. Acad. Sci. USA. 98:12527-12531.

Related Resources:

Review of antibody production and immunogen preparation methods
Review of sulfhydryl-reactive (maleimide) crosslinker chemistry
Carrier protein activation and conjugation data for immunogen preparation

Related Products:

SulfoLink Immobilization Kit for Peptides – for preparing an affinity column with a cysteine peptide
Keyhole Limpet Hemocyanin (non-activated KLH) – for conjugation through amines or carboxylates
BSA and other Carrier Proteins


Ordering Information
 
Product # Description Pkg. Size Instructions MSDS CofA Price
77606 Imject Maleimide-Activated mcKLH
Formulation: Activated protein lyophilized in 10mM sodium phosphate, 150mM sodium chloride, 10mM EDTA, 50mM sucrose, pH 7.2
Sufficient For: 1 conjugation reaction with up to 2mg sulfhydryl-containing peptide
2mg Product Instructions for product #77606 Imject Maleimide-Activated mcKLH MSDS for product #77606 Imject Maleimide-Activated mcKLH Certificate of Analysis for product #77606 Imject Maleimide-Activated mcKLH $68.00

Add to cart
77605 Imject Maleimide-Activated mcKLH
Formulation: Activated protein lyophilized in 10mM sodium phosphate, 150mM sodium chloride, 10mM EDTA, 50mM sucrose, pH 7.2
Sufficient For: 5 conjugation reactions, each with up to 2mg sulfhydryl-containing peptide
10mg Product Instructions for product #77605 Imject Maleimide-Activated mcKLH MSDS for product #77605 Imject Maleimide-Activated mcKLH Certificate of Analysis for product #77605 Imject Maleimide-Activated mcKLH $313.00

Add to cart
77610 Imject Maleimide-Activated mcKLH
Formulation: Activated protein lyophilized in 10mM sodium phosphate, 150mM sodium chloride, 10mM EDTA, 50mM sucrose, pH 7.2
Sufficient For: 50 conjugation reactions, each with up to 2mg sulfhydryl-containing peptide
10 x 10mg Product Instructions for product #77610 Imject Maleimide-Activated mcKLH MSDS for product #77610 Imject Maleimide-Activated mcKLH $2,502.00

Add to cart
77605B Imject Maleimide-Activated mcKLH
Formulation: Activated protein lyophilized in 10mM sodium phosphate, 150mM sodium chloride, 10mM EDTA, 50mM sucrose, pH 7.2
Custom MSDS for product #77605B Imject Maleimide-Activated mcKLH Certificate of Analysis for product #77605B Imject Maleimide-Activated mcKLH
77164 Imject Maleimide Conjugation Buffer
Formulation: 0.083M sodium phosphate, 0.1M EDTA, 0.9M NaCl, 0.1M sorbitol and 0.02% sodium azide; pH 7.2
Sufficient For: Dissolving up to 60mg of sulfhydryl-peptide for maleimide conjugation with carrier proteins
30mL MSDS for product #77164 Imject Maleimide Conjugation Buffer $39.00

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77666 Imject Maleimide-Activated mcKLH Spin Kit
Formulation: Multi-component kit
Sufficient For: 5 conjugation reactions and complete immunogen preparations, each with up to 2mg sulfhydryl-containing peptide

Kit Contents:
Imject Maleimide-Activated mcKLH, 5 x 2mg
Imject Maleimide Conjugation Buffer, 30mL
Imject Purification Buffer Salts, 5 x 0.84g
Zeba Spin Desalting Columns, 7K MWCO, 2mL, 5 columns
5 x 2mg kit Product Instructions for product #77666 Imject Maleimide-Activated mcKLH Spin Kit MSDS for product #77666 Imject Maleimide-Activated mcKLH Spin Kit $402.00

Add to cart
77663 Imject Maleimide PEGylated mcKLH
Formulation: Activated protein lyophilized in 10mM sodium phosphate, 150mM sodium chloride, 10mM EDTA, 50mM sucrose, pH 7.2
Sufficient For: 5 conjugation reactions, each with up to 2mg sulfhydryl-containing peptide
10mg Product Instructions for product #77663 Imject Maleimide PEGylated mcKLH MSDS for product #77663 Imject Maleimide PEGylated mcKLH $323.00

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Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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