Protein Labeling Kit

Products - Amplify & Label - Protein Labeling Kits

Featuring ArrayIt® Green540 and ArrayIt® Red640 dyes. ArrayIt® Green540 and ArrayIt® Red640 are new dyes providing superior labeling efficiently and photo bleaching qualities over the current popular dyes used in the microarray industry. ArrayIt® Red640 matches perfectly current microarray laser and filter sets for Cy5 absorption and emission spectrum and ArrayIt® Green540 perfectly for Cy3.

ArrayIt’s fluorescent labeling kit for proteins includes everything needed for high quality direct labeling of proteins for protein microarray binding reactions including fluorescent dyes, reagents and purification columns. This kit takes advantage of an efficient direct labeling to produce fluorescently labeled proteins for use in microarray reactions. This kit produces higher yield and much more efficiently labeled proteins than other popular labeling approaches.

Whether you are labeling monoclonal antibodies, secondary IgG, enzymes or other proteins, these kits will provide unsurpassed in simplicity, ease-of-use and flexibility.

Our amine reactive dyes are excellent alternatives to the Cy3 and Cy5 Dyes used in conjunction with today’s most popular microarray scanners. The absorption for ArrayIt Red is 540 with an emission of 570. ArrayIt Green absorbs at 640 and emits at 670. It is not necessary to purchase new filter sets to use these kits.

Table of Contents

Introduction
Congratulations on taking a big step towards improving the economies of scale, quality and speed of your genomics research. This booklet contains a complete set of protocols including the steps and principles needed to use the 2-Color Fluorescent Labeling Kit for proteins.

Quality Control
TeleChem assures the performance of these products. The finest scientific research went into the development of these products. Rigorous quality control monitoring on a lot-by-lot basis guarantees that the columns, dyes, buffers, and protocols conform to the highest industry standards.

Product Description
This product is designed to use in conjunction with protein microarray manufactured by ArrayIt. However, it is compatible with other downstream applications.

Users will appreciate the following features:

Supports all substrate and slide surface chemistries
Easy protocol, high yields and efficient incorporation
Bright photo stable dyes
Can be used with both two- and three-dimensional surfaces
Superior separation chemistry provides 99+% probe purity
Dye, hapten and salt removal reduces background
Kits arrive ready to use, no buffer or column preparation required
Microcentrifuge format allows rapid purification with single columns
90% recovery
Consistent results
Easy to use

Short Protocol 1 mg protein

Dissolve protein 1 mg per 100 l in PBS in a reaction tube.
Add 20 l of reaction solution A to the protein reaction tube.
Prepare the reactive ArrayIt^® Green540 and ArrayIt^® Red640 stock just prior to starting the reaction. Add 25 l of solution B to 1 tube of dye and pipette up and down to mix and dissolve.
Combine 10 l of reactive dye solution to the protein reaction tube and vortex gently to mix. Remaining dye solution should be stored at 4C and can be used once more within 1 week.
Let reaction run for 1 hour at room temperature protected from light
While reaction runs, prepare two purification columns.
Gently tap columns to insure that all the gel is at the bottom of the column. Hydrate dry gel in purification column by adding 0.8 ml of solution C to each column and vortex vigorously for ~5 seconds. Remove any air bubbles by tapping the bottoms of the columns sharply. Wait 30 minutes before using, drain excess fluid from column prior to use.
Stop dye labeling reaction using buffer D (30 minutes).
Purify reaction by transferring half of the protein reaction tube contents to two purification columns and spin at 750 x g for 2 minutes to collect dye conjugated proteins.

Complete Protocol

Figure 1. Two color antibody microarray. Compare two biological samples to measure the absolute and relative differences in protein expression. This procedure is fluorescence-based and compatible with all current microarray scanners. Extracted proteins are directly labeled (kit provided) and analyzed. The covalently immobilized antibodies capture fluorescently labeled antigens during the reaction step. Normal and cancerous cells can be compared. The raw data provide a measure of proteins from samples A & B (see above graphic).

Technical Assistance
Please contact us if you have any comments, suggestions, or if you need technical assistance. By electronic mail: arrayit@arrayit.com (under the subject heading please type ArrayIt technical assistance). By telephone: (408) 744-1331, Monday—Friday PST 9:00am - 4:30pm. Please remember that we want to hear about your successes!

Equipment and Reagents
Antibody Micorarrays
Super Microarray Substrates
Mirror Substrates
Hybridization Solutions
Hybridization Cassettes
Microarray Instruments

Troubleshooting Tips
Dyes, haptens, chromophores and salts can be removed from proteins other than IgG using Pro·Spin spin columns. However, recovery of the protein may be reduced due to specific or non-specific interactions with the column matrix. Below is a list of common modes of interaction together with suggestions for their minimization.

Problem & Possible Solution
Hydrophobic interaction with column matrix

Add non-ionic detergent, decrease ionic strength or increase pH of hydration buffer

Ionic interaction with column matrix

Increase ionic strength of hydration buffer

Sample precipitation

Increase ionic strength, remove salt from hydration buffer or dilute sample.

Protease degradation of sample

Add suitable protease inhibitor to hydration buffer.

Kit Contents

Ordering Information

Product

Description

Catalog ID

Price per kit (US dollars)*

2 color fluorescent Labeling Kit for proteins

Inquire

Inquire

Discounts:
10-19 kits, 10% discount
20-50 kits, 15% discount
50+ kits, 20% discount

*To order ArrayIt® Brand Products: call 408-744-1331, fax 408-744-1711 or click on the button below to proceed directly to the purchase page.

*International pricing may vary as much as 20% due to import duties, stocking fees and technical support.

Storage Conditions
ArrayIt® Red and Green should be stored at –20°C. Fluorescent Probe Purification columns and reagents can be stored dry at room temperature (20-25°C). The kits perform well across a wide range of ambient temperatures and relative humidity. The kits are nuclease-free, sterile, and have a shelf life of one-year from the date of purchase.

Warranty
ArrayIt® brand products have been scientifically developed and are sold for research purposes only. ArrayIt® brand products are not for use in human diagnostics or for drug purposes, nor are they to be administered to humans in any way. Extreme care and exact attention should be practiced in the use of the materials described herein. All ArrayIt® brand products are subject to extensive quality control and are guaranteed to perform as described when used properly. Any problems with our ArrayIt® brand product should be reported to TeleChem immediately. TeleChem’s liability is limited to the replacement of the product, or a full refund. Any misuse of this product including deviations from our protocols is the full responsibility of the user, and TeleChem makes no warranty or guarantee under such circumstances.

Cy3 and Cy5 are trademarks of Amersham Corporation.