Invitrogen™ Click-iT™ Protein Enrichment Kit, for click chemistry capture of azide-modified proteins

Catalog No.	Quantity
C10416	1 Kit

Catalog No. C10416 Supplier Invitrogen™ Supplier No. C10416

Description

Capture of click-chemistry-based azide-modified proteins

The Click-iT Protein Enrichment Kit for the efficient capture of click chemistry based azide-modified proteins on a resin of alkyne agarose. Superior to biotin or lectin based enrichment approaches. Ideal for proteomics, biomarker discovery, post-translational modification (PTM) analysis, and more. The azide modification can occur via metabolic feeding, enzymatic addition, or chemical modification. Click-Azide modified proteins, or their post-translationally modified forms, are enriched from complex protein extracts on the alkyne-agarose resin supplied. Once anchored to the resin via copper catalyzed click chemistry, extensive washing yields a highly enriched population of nascent molecules that can be furthered characterized by mass spectrometry. The alkyne-agarose resin improves upon existing biotin approaches by >8 fold, with signal to noise of biotin = 3 while the Click-iT resin = 25. Perfect upstream MS-based sample preparation technique for looking at changes in global protein expression and biomarker analysis.

• Global differential profiling of multiple subclasses of posttranslationally modified (PTM) proteins and newly synthesized proteins
• Improved signal to noise by eliminating nonspecific binding and increasing selectivity, thus improving detection of low abundant proteins
• Accelerates PTM site identification leading to more rapid initiation of functional studies
• Fully compatible with widely used MS techniques including iTRAC and ICAT
• Seamless integration of cell biology with protein chemistry
• Affordable, easy, no special equipment required
• One Reaction, Endless Possibilities

Cell Analysis, Click Chemistries, Labeling Chemistry, Mass Spectrometry, Metabolic Labeling and Nascent Protein Synthesis with Click Chemistry, Protein and Peptide Purification, Protein Enrichment, Protein Sample Preparation and Protein Purification, Proteins, Expression, Isolation and Analysis, Spectrometry Sample Preparation Reagents

Order Info

Shipping Condition: Room temperature

Specifications

• Product Type: Protein Enrichment Kit
• Quantity: 1 Kit
• Content And Storage: Store at 2°C to 8°C. Desiccation recommended. Protect material from long-term exposure to light; may be exposed to light for short periods of time
• Shipping Condition: Room Temperature
• Product Line: Click-iT
• Format: Spin Column

Frequently Asked Questions (FAQs)

What is the expected peptide yield for samples prepared with the Click-iT Protein Enrichment Kit, for click chemistry capture of azide-modified proteins (Cat. No. C10416)?

The yield of purified peptides from samples prepared with the Click-iT Protein Enrichment Kit, for click chemistry capture of azide-modified proteins (Cat. No. C10416), strongly depends on the sample type, the incorporation efficiency of the azide analog used, and the amount of lysate added to the resin. In the examples presented in the posters linked below, we were able to recover approximately 25-50 µg of purified peptides from about 0.5 to 8 mg of cell lysate, using approximately 100 µL of resin.

Click chemistry enrichment of cell surface glycoproteins

A universal click chemistry-based enrichment column for the identification of multiple subclasses of PTM-modified proteins and newly synthesized proteins

How can I confirm the protein is enriched in samples prepared with the Click-iT Protein Enrichment Kit, for click chemistry capture of azide-modified proteins (Cat. No. C10416)?

To confirm the protein is enriched in samples prepared with the Click-iT Protein Enrichment Kit, for click chemistry capture of azide-modified proteins (Cat. No. C10416) we recommend saving the flow through from the column along with a small aliquot of the initial cell lysate and running these on a gel together. This will show the depletion of the azide-labeled proteins from the lysate. Please refer to the posters below that show examples of how we have analyzed peptide recovery (or depletion of the lysate):

Click chemistry enrichment of cell surface glycoproteins

A universal click chemistry-based enrichment column for the identification of multiple subclasses of PTM-modified proteins and newly synthesized proteins

For Research Use Only. Not for use in diagnostic procedures.