Protein Citrullination alters protein structure and function, playing a critical role in normal physiological processes like gene regulation and skin keratinization, as well as the pathogenesis of autoimmune diseases like rheumatoid arthritis. RayBio^® Human Protein Citrullination Antibody Array 8000 is specifically designed for simultaneous identification of the relative levels of citrullination of 8000 different human proteins in cell lysate, culture supernatant, serum, plasma and other biological samples. By monitoring the changes in protein citrullination in your experimental model system, you can verify oxidative damage without spending excess time and effort performing an analysis of immunoprecipitation and/or Western Blot.

RayBio^® Human Protein Citrullination Antibody Array utilizes the ‘biotin citrulline probes’ method to biotin label citrulline groups of citrullinated proteins. In this method, the biotin labeled phenylglyoxal group of the probe selectively labels citrulline of the sample at acidic pH.. The biotin labeled sample then is added into antibody array glass slide wells. The antibody array slide wells are washed. After incubation with a fluorescent dye-conjugated streptavidin (Cy3 equivalent), the slides can then be imaged using a laser scanner, such as the Axon GenePix, using the Cy3 channel.

• High sensitivity and specificity
• Low sample volume (10-100 µL per array)
• Large dynamic range of detection
• Compatible with most sample types
• Test 4 or 8 samples on each slide
• Suitable for high-throughput assays

• Human Antibody Array Slide(s)
• Blocking Buffer
• Fluorescent Dye-Conjugated Streptavidin
• Wash Buffer I
• Wash Buffer II
• Wash Buffer III
• Lysis Buffer
• Protease Inhibitor Cocktail
• Citrullination Buffer
• Citrullination Labeling Reagent
• Citrullination Quenching Buffer
• Adhesive film

• Trichloroacetic acid, ≥ 99% (hazardous)
• Acetone, ≥ 98% (hazardous)
• 1.5 mL microcentrifuge tubes
• 15 mL tubes (polypropylene)
• 10 mL graduated cylinders
• Benchtop centrifuge and microcentrifuge
• Precision pipettes to deliver 2 µL to 1 mL volumes
• Adjustable 1-25 mL pipettes for reagent preparation
• Water bath or heat block
• Shaker
• Laser scanner for fluorescence detection
• Aluminum foil
• Distilled water
• Plastic box
• 50 mL Centrifuge tube
• Isopropanol (2-propanol)

1. Dry the glass slide
2. Add Blocking Buffer to each well and incubate.
3. Add the sample to each well and incubate.
4. Wash the wells.
5. Add Fluorescent Dye-Conjugated Streptavidin solution to each well and incubate.
6. Wash the wells.
7. Scan with a gene microarray laser scanner
8. Perform densitometry and analysis