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Protein A product

Detection Reagents

Protein A-HiLyte™ Fluor conjugates are an excellent alternative to dye-labeled secondary antibodies. These highly efficient conjugates constitute a series of universal detection reagents, which combine the binding capacity of Protein A with the high fluorescent intensity of HiLyte™ Fluor dyes.

Benefits

  • Universal: only one reagent to detect multiple targets.
  • Large panel of emission wavelength: HiLyte™ Fluor fluorescence emission spans the full visible and near infrared red spectra.
  • Super bright signal: HiLyte™ Fluor conjugates exhibit more intense fluorescence compared to other spectrally similar classical dyes (TAMRA, FITC,...).


DescriptionSize Reference USD Qty  
Protein A, recombinant5 mg AS-72233-5 77.00Order Form
Protein A, recombinant25 mg AS-72233-25 154.00Order Form
Protein A, recombinant100 mg AS-72233-100 360.00Order Form
Protein A, recombinant1000 mg AS-72233-1000 1 200.00Order Form
Protein A-Biotin Conjugate5 mg AS-72234 154.00Order Form
Protein A-HiLyte™ Fluor 488 Conjugate1 mg AS-72235 132.00Order Form
Protein A-HiLyte™ Fluor 555 Conjugate1 mg AS-72236 132.00Order Form
Protein A-HiLyte™ Fluor 647 Conjugate1 mg AS-72237 132.00Order Form
Protein A-HiLyte™ Fluor 750 Conjugate1 mg AS-72239 132.00Order Form

Protein A-HiLyte™ Fluor conjugates are an excellent alternative to dye-labeled secondary antibodies. These highly efficient conjugates constitute a series of universal detection reagents, which combine the binding capacity of Protein A with the high fluorescent intensity of HiLyte™ Fluor dyes.

  • Protein A is a non-glycosylated cell wall protein of Staphylococcus aureus.1-4
  • It consists of five IgG binding domains (A, B, C, D, and E), each is approximately 60 amino acids long with no cysteines present and two Staphylococcus aureus cell membrane binding domains (X and M).3
  • It can bind with strong affinity the Fc portion of immunoglobulin of certain species (See Table below for affinity strength).1-4
Species
Immunoglobulin
Protein A Binding
Human
Human IgG1, IgG2, IgG4
Strong
Human IgM, IgA
Weak
Mouse
Mouse IgG1
Weak
Mouse IgG2A, IgG2B, IgG3
Strong
Rat
Rat IgG1
Weak
Rat IgG2C
Strong
Goat
Goat IgG1
Weak
Goat IgG2
Strong
Bovine
Bovine IgG2
Strong
Guinea Pig
IgG
Strong
Rabbit
IgG
Strong
  • Its apparent molecular weight is approximately 30,000 Da compared to 42,000 Da for the native Protein A found in Staphylococcus aureus.
  • Protein A is available as unlabeled or labeled with Biotin.
  • Protein A is also available as HiLyte™ Fluor dye labeled:
    • Protein A-H488 (Ex/Em = 499/523 nm)
    • Protein A-H555 (Ex/Em = 553/568 nm)
    • Protein A-H647 (Ex/Em = 649/674 nm)
    • Protein A-H750 (Ex/Em = 754/778 nm)

Applications

  • HiLyte™ Fluor labeled Protein A conjugates can be used as a universal reagent to detect primary antibodies in Western Immunoblot from many species including rabbit, human, some mouse IgG isotypes, and others.
  • Unlabeled Protein A can be used for immunoprecipitation, antibodies purification, and assay development

3T3 cells were stained with Rabbit-anti-tubulin IgG (0.5 μg/mL) followed by Protein A-HiLyteTM Fluor 488 (2), Protein A-HiLyteTM Fluor 555 (3) and Protein A-HiLyteTM Fluor 647 (1) at 0.5 μg/mL.

Rabbit anti-Actin antibody was probed with ProteinA-HiLyte™ Fluor 680 and ProteinA-HiLyte™ Fluor 750 conjugates.

HiLyte™ Fluor 680 HiLyte™ Fluor 750  

References

  1. Björk, I. Eur J Biochem 29 (1972):579-584.
  2. Sjödahl, J. Eur J Biochem 78 (1977): 471-490.
  3. Schröder, A. K. et al. Immuno l57 (1985): 305-309.
  4. Rosenberg, I.M. Protein Analysis and Purification, Birkhäuser, 2005: 41-47.
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