Actively helping customers, employees and the global community during the coronavirus SARS-CoV-2 outbreak.  Learn more >>
Actively helping customers, employees and the global community during the coronavirus SARS-CoV-2 outbreak.  Learn more >>
logo
AAT Bioquest

ReadiLink™ BSA Conjugation Kit

EDC reacts with a carboxyl group of carrier protein BSA or KLH (represented by the red ball), forming an amine-reactive O-acylisourea intermediate (the central molecule). The O-acylisourea intermediate reacts with an amine group on the antigen molecule represented by the smaller blue ball, yielding a conjugate of the two molecules joined by a stable amide bond [Please note the O-acylisourea inermediate is also susceptible to hydrolysis, making it unstable and short-lived in aqueous solution].
EDC reacts with a carboxyl group of carrier protein BSA or KLH (represented by the red ball), forming an amine-reactive O-acylisourea intermediate (the central molecule). The O-acylisourea intermediate reacts with an amine group on the antigen molecule represented by the smaller blue ball, yielding a conjugate of the two molecules joined by a stable amide bond [Please note the O-acylisourea inermediate is also susceptible to hydrolysis, making it unstable and short-lived in aqueous solution].
EDC reacts with a carboxyl group of carrier protein BSA or KLH (represented by the red ball), forming an amine-reactive O-acylisourea intermediate (the central molecule). The O-acylisourea intermediate reacts with an amine group on the antigen molecule represented by the smaller blue ball, yielding a conjugate of the two molecules joined by a stable amide bond [Please note the O-acylisourea inermediate is also susceptible to hydrolysis, making it unstable and short-lived in aqueous solution].
Ordering information
Price
Catalog Number
Unit Size
Quantity
Add to cart
Additional ordering information
Telephone1-800-990-8053
Fax1-800-609-2943
Emailsales@aatbio.com
InternationalSee distributors
ShippingStandard overnight for United States, inquire for international
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
UNSPSC12352200
Related products
ReadiLink™ Rapid mFluor™ Violet 450 Antibody Labeling Kit *Microscale Optimized for Labeling 50 μg Antibody Per Reaction*
ReadiLink™ Rapid mFluor™ Violet 420 Antibody Labeling Kit *Microscale Optimized for Labeling 50 μg Antibody Per Reaction*
ReadiLink™ Rapid mFluor™ Violet 510 Antibody Labeling Kit *Microscale Optimized for Labeling 50 μg Antibody Per Reaction*
ReadiLink™ Rapid mFluor™ Violet 540 Antibody Labeling Kit *Microscale Optimized for Labeling 50 μg Antibody Per Reaction*
ReadiLink™ Rapid mFluor™ Blue 570 Antibody Labeling Kit *Microscale Optimized for Labeling 50 μg Antibody Per Reaction*
ReadiLink™ Rapid mFluor™ Green 620 Antibody Labeling Kit *Microscale Optimized for Labeling 50 μg Antibody Per Reaction*
ReadiLink™ Rapid mFluor™ Yellow 630 Antibody Labeling Kit *Microscale Optimized for Labeling 50 μg Antibody Per Reaction*
ReadiLink™ Rapid mFluor™ Red 700 Antibody Labeling Kit *Microscale Optimized for Labeling 50 μg Antibody Per Reaction*
ReadiLink™ Rapid mFluor™ Red 780 Antibody Labeling Kit *Microscale Optimized for Labeling 50 μg Antibody Per Reaction*
ReadiLink™ Rapid iFluor® 350 Antibody Labeling Kit *Microscale Optimized for Labeling 50 μg Antibody Per Reaction*
ReadiLink™ Rapid iFluor® 555 Antibody Labeling Kit *Microscale Optimized for Labeling 50 μg Antibody Per Reaction*
ReadiLink™ Rapid iFluor® 594 Antibody Labeling Kit *Microscale Optimized for Labeling 50 μg Antibody Per Reaction*
ReadiLink™ Rapid iFluor® 647 Antibody Labeling Kit *Microscale Optimized for Labeling 50 μg Antibody Per Reaction*
ReadiLink™ Rapid iFluor® 680 Antibody Labeling Kit *Microscale Optimized for Labeling 50 μg Antibody Per Reaction*
ReadiLink™ Rapid iFluor® 700 Antibody Labeling Kit *Microscale Optimized for Labeling 50 μg Antibody Per Reaction*
ReadiLink™ Rapid iFluor® 750 Antibody Labeling Kit *Microscale Optimized for Labeling 50 μg Antibody Per Reaction*
ReadiLink™ Rapid iFluor® 488 Antibody Labeling Kit *Microscale Optimized for Labeling 50 μg Antibody Per Reaction*
ReadiLink™ Rapid iFluor® 633 Antibody Labeling Kit *Microscale Optimized for Labeling 50 μg Antibody Per Reaction*
ReadiLink™ Rapid iFluor® 790 Antibody Labeling Kit *Microscale Optimized for Labeling 50 μg Antibody Per Reaction*
ReadiLink™ Rapid FITC Antibody Labeling Kit *Microscale Optimized for Labeling 50 μg Antibody Per Reaction*
ReadiLink™ Rapid trFluor™ Eu Antibody Labeling Kit *Microscale Optimized for Labeling 50 ug Antibody Per Reaction*
ReadiLink™ Rapid trFluor™ Tb Antibody Labeling Kit *Microscale Optimized for Labeling 50 ug Antibody Per Reaction*
ReadiLink™ Protein Conjugation Stop Buffer
ReadiLink™ Protein Biotinylation Kit
ReadiLink™ Protein Biotinylation Kit *Powered by ReadiView™ Biotin Visionization Technology*
ReadiLink™ Rapid Cy3 Antibody Labeling Kit *Microscale Optimized for Labeling 50 μg Antibody Per Reaction*
ReadiLink™ Rapid Cy5 Antibody Labeling Kit *Microscale Optimized for Labeling 50 μg Antibody Per Reaction*
ReadiLink™ Rapid Cy7 Antibody Labeling Kit *Microscale Optimized for Labeling 50 μg Antibody Per Reaction*
ReadiLink™ xtra Rapid iFluor® 350 Antibody Labeling Kit *BSA-Compatible*
ReadiLink™ xtra Rapid iFluor® 488 Antibody Labeling Kit *BSA-Compatible*
ReadiLink™ xtra Rapid iFluor® 555 Antibody Labeling Kit *BSA-Compatible*
ReadiLink™ xtra Rapid iFluor® 594 Antibody Labeling Kit *BSA-Compatible*
ReadiLink™ xtra Rapid iFluor® 647 Antibody Labeling Kit *BSA-Compatible*
ReadiLink™ xtra Rapid iFluor® 750 Antibody Labeling Kit *BSA-Compatible*
ReadiLink™ xtra Rapid FITC Antibody Labeling Kit *BSA-Compatible*
ReadiLink™ xtra Rapid Cy3 Antibody Labeling Kit *BSA-Compatible*
ReadiLink™ xtra Rapid Cy5 Antibody Labeling Kit *BSA-Compatible*
ReadiLink™ xtra Rapid Cy7 Antibody Labeling Kit *BSA-Compatible*
ReadiLink™ xtra Rapid XFD594 Antibody Labeling Kit *BSA-Compatible, XFD594 Same Structure to Alexa Fluor™ 594*
ReadiLink™ Rapid XFD488 Antibody Labeling Kit *XFD488 Same Structure to Alexa Fluor™ 488*
ReadiLink™ Rapid XFD555 Antibody Labeling Kit *XFD555 Same Structure to Alexa Fluor™ 555*
ReadiLink™ Rapid XFD594 Antibody Labeling Kit *XFD594 Same Structure to Alexa Fluor™ 594*
ReadiLink™ Rapid XFD647 Antibody Labeling Kit *XFD647 Same Structure to Alexa Fluor™ 647*
ReadiLink™ Rapid XFD750 Antibody Labeling Kit *XFD750 Same Structure to Alexa Fluor™ 750*
ReadiLink™ iFluor® 488 Nick Translation dsDNA Labeling Kit
ReadiLink™ iFluor® 555 Nick Translation dsDNA Labeling Kit
ReadiLink™ iFluor® 647 Nick Translation dsDNA Labeling Kit
ReadiLink™ Cy3 Nick Translation dsDNA Labeling Kit
ReadiLink™ Cy5 Nick Translation dsDNA Labeling Kit
ReadiLink™ Biotin Nick Translation dsDNA Labeling Kit
ReadiLink™ DIG (Digoxigenin) Nick Translation dsDNA Labeling Kit
ReadiLink™ iFluor® 488 Oligo and ssDNA Labeling Kit
ReadiLink™ iFluor® 555 Oligo and ssDNA Labeling Kit
ReadiLink™ iFluor® 647 Oligo and ssDNA Labeling Kit
ReadiLink™ Cy3 Oligo and ssDNA Labeling Kit
ReadiLink™ Cy5 Oligo and ssDNA Labeling Kit
ReadiLink™ Biotin Oligo and ssDNA Labeling Kit
ReadiLink™ DIG (Digoxigenin) Oligo and ssDNA Labeling Kit
ReadiLink™ xtra Rapid XFD488 Antibody Labeling Kit *BSA-Compatible, XFD488 Same Structure to Alexa Fluor™ 488*
ReadiLink™ xtra Rapid XFD555 Antibody Labeling Kit *BSA-Compatible, XFD555 Same Structure to Alexa Fluor™ 555*
ReadiLink™ xtra Rapid XFD647 Antibody Labeling Kit *BSA-Compatible, XFD647 Same Structure to Alexa Fluor™ 647*
ReadiLink™ xtra Rapid XFD750 Antibody Labeling Kit *BSA-Compatible, XFD750 Same Structure to Alexa Fluor™ 750*
ReadiLink™ iFluor® 488 FISH Fluorescence Imaging Kit
Show More (53)

OverviewpdfSDSpdfProtocol


Bovine serum albumin (BSA) is a serum albumin protein that has numerous biochemical applications including ELISAs (Enzyme-Linked Immunosorbent Assay), immunoblots, and immunohistochemistry. Like most abundant plasma proteins, it is very stable and soluble. In addition, the 67 kDa protein is sufficiently large and complex to be fully immunogenic. It contains numerous sites per molecule for effective conjugation of peptides and other antigens using amine-reactive or carboxyl-reactive crosslinkers. Consequently, BSA is a popular carrier protein for conjugation to haptens and other weak antigens to make them more immunogenic for antibody production. This ReadiLink™ BSA Conjugation kit is primarily optimized for the simple preparation of hapten-carrier conjugates for immunization and antibody production. The ReadiLink™ BSA Conjugation kit is one-step conjugation of a hapten to a carrier protein using the carboxyl-reactive carbodiimide as the crosslinker. The resulting conjugate is used for eliciting an immune response and antibody production against the hapten. The carboxyl-reactive carbodiimide reacts with exposed carboxyl and amino groups on peptides and proteins to form stable bonds. These kits contain BSA formulated in buffers compatible with the carboxyl-reactive carbodiimide reactions and desalt spin columns, which offer exceptional protein recovery by simple centrifugation step.

Components


Example protocol


AT A GLANCE

Protocol Summary
  1. Prepare protein solution
  2. Prepare hapten solution
  3. Mix protein with hapten into EDC
  4. Incubate the reaction at RT for 2 hrs
  5. Purify the conjugate by desalting 

Important
The following protocol is a general protocol for a wide variety of haptens. Optimize the protocol accordingly for the conjugation efficiencies upon the size and structure of your hapten. Using a molar excess of hapten over carrier protein ensures efficient conjugation. In general, a reaction with equal mass amounts of hapten and carrier protein will achieve sufficient molar excess.

SAMPLE EXPERIMENTAL PROTOCOL

Prepare BSA-Hepten Conjugation:
  1. Add 200 µL of ddH2O into the vial of BSA (Component A) to make a 10 mg/mL solution.
  2. Dissolve up to 2 mg hapten in 450 µL Conjugation Buffer (Component B).
    Note     Some haptens might have limited solubility, use DMSO (<30% in the final conjugation solution) to dissolve it first. Higher concentration of DMSO might irreversibly denature the carrier protein.
  3. Add the 450 µL hapten solution into the 200 µL BSA solution to have Hapten-BSA mixture solution.
  4. Add the Hapten-BSA solution into one vial of EDC (10mg) (Component C), dissolve it by gentle mixing. Incubate at room temperature for 2 hours.
  5. Purify the conjugate by desalting to remove non-reacted crosslinker and protein preservative (e.g., sodium azide). 

Purify BSA-Hepten conjugate:
  1. Thaw 1 bottle of Purification Buffer (Component D) to room temperature before use. Unused buffer can be stored at 4 °C for 1 week.
  2. Twist off the bottom closure of the desalting column (Component E) and loosen the cap. Place the column in a collection tube.
  3. Centrifuge the column at 1,000g for 2 minutes to remove the storage solution.
  4. Remove the cap and slowly add 1 mL of purification buffer to the column. Centrifuge at 1,000g for 2 minutes, remove the buffer. Repeat this step for 3 additional times, discarding the buffer from the collection tube.
  5. Place the column to a new collection tube, and gently apply the sample into the center of the compact resin bed.
  6. Centrifuge the column at 1,000g for 2 minutes to collect the sample.
  7. The Hapten-BSA conjugate can now be used for immunization. If the Hapten-BSA conjugate is to be stored for more than a few days, sterile filter the conjugate, and store at 4 °C or -20 °C.
    Note     If the conjugate is to be used within one week, PBS may be used for purification. If the conjugate is to be frozen, use the purification buffer salts (Component D) for purification. If DMSO is used in the conjugation, prepare the purification buffer salts with the same percentage of DMSO used for conjugation. This will minimize the precipitation in the column during desalting. If a precipitate formed during conjugation, centrifuge the precipitated material, collect the supernatant and save the precipitate. Purify the supernatant. Combine the precipitate and the purified conjugate. 

Product Family


Images


References


View all 29 references: Citation Explorer
Optimization of the preparation process of vinblastine sulfate (VBLS)-loaded folate-conjugated bovine serum albumin (BSA) nanoparticles for tumor-targeted drug delivery using response surface methodology (RSM)
Authors: Zu Y, Zhang Y, Zhao X, Zhang Q, Liu Y, Jiang R.
Journal: Int J Nanomedicine (2009): 321
Characterization of bovine serum albumin glycated with glucose, galactose and lactose
Authors: Ledesma-Osuna AI, Ramos-Clamont G, Vazquez-Moreno L.
Journal: Acta Biochim Pol (2008): 491
Improvement of functional properties of bovine serum albumin through phosphorylation by dry-heating in the presence of pyrophosphate
Authors: Enomoto H, Li CP, Morizane K, Ibrahim HR, Sugimoto Y, Ohki S, Ohtomo H, Aoki T.
Journal: J Food Sci (2008): C84
Polyethylene glycol improves conjugation of bovine hemoglobin and human serum albumin in a controlled ratio
Authors: Zheng C, Bi J, Ma G, Su Z.
Journal: Artif Cells Blood Substit Immobil Biotechnol (2007): 568
Purification of denatured bovine serum albumin coated CdTe quantum dots for sensitive detection of silver(I) ions
Authors: Wang JH, Wang HQ, Zhang HL, Li XQ, Hua XF, Cao YC, Huang ZL, Zhao YD.
Journal: Anal Bioanal Chem (2007): 969
Effect of lyophilization on the structure and phase changes of PEGylated-bovine serum albumin
Authors: Tattini V, Jr., Parra DF, Polakiewicz B, Pitombo RN.
Journal: Int J Pharm (2005): 124
Preparation of bovine serum albumin nanoparticles surface-modified with glycyrrhizin
Authors: Mao SJ, Hou SX, Zhang LK, Jin H, Bi YQ, Jiang B.
Journal: Yao Xue Xue Bao (2003): 787
Products of spontaneous conjugation of aflatoxins with bovine serum albumin: immunochemical properties
Authors: Burkin AA, Kononenko GP, Soboleva NA.
Journal: Prikl Biokhim Mikrobiol (2003): 228
Generation of polyclonal antibody against mu-conotoxin GIIIA using an immunogen of [Cys(5)]mu-conotoxin GIIIA site-specifically conjugated with bovine serum albumin
Authors: Nakamura M, Oba Y, Mori T, Sato K, Ishida Y, Matsuda T, Nakamura H.
Journal: Biochem Biophys Res Commun (2002): 1037
Mice vaccinated with the O-antigen of Francisella tularensis LVS lipopolysaccharide conjugated to bovine serum albumin develop varying degrees of protective immunity against systemic or aerosol challenge with virulent type A and type B strains of the patho
Authors: Conlan JW, Shen H, Webb A, Perry MB.
Journal: Vaccine (2002): 3465