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Invitrogen™ ZOOM™ Carrier Ampholytes 4-6, 10mL

Catalog No. ZM0025
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ZM0025 10 mL
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Catalog No. ZM0025 Supplier Invitrogen™ Supplier No. ZM0025
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Description

Small, soluble molecules with both positive and negative charge groups

They sort based on their isoelectric points in an electric field, thereby establishing a pH gradient in solution or contributing to the gradient when used with IPG strips. Carrier ampholytes help stabilize the pH gradient and current in IPG strips and aid in protein solubility, resulting in reproducible IEF resolution.

Order Info

Shipping Condition: Room Temperature

Specifications

Content And Storage ZOOM™ Carrier Ampholytes are supplied as 10 ml bottles. Store at +4°C. Guaranteed stable for 1 year when properly stored.
pH Range 4 to 6
Quantity 10 mL
Shipping Condition Room Temperature
Product Line ZOOM
Product Type Carrier Ampholyte

Frequently Asked Questions (FAQs)

When running the ZOOM IEF Fractionator, if there is not a solubility issue, will the proteins/peptides in the sample migrate to their expected chambers based on isoelectric point in the absence of carrier ampholytes? Won't they act as ampholytes within each chamber?

Proteins are ampholytes, but in general they are poor carrier ampholytes. The profiles of the fractions are similar, but not identical, to those obtained with ampholytes in the sample.
Can I heat my protein sample prepared using ZOOM 2D Protein Solubilizer?

We do not recommend heating protein samples containing urea over 37 degrees C as elevated temperatures cause urea to hydrolyze to isocyanate, which modifies proteins by carbamylation.

How do you recommend storing protein samples prepared in the Sample Rehydration buffer?

We recommend storing them at -80 degrees C. We do not recommend storing them at -20 degrees C
How much ampholyte do you recommend adding to the sample rehydration buffer?

The recommended ampholyte concentration in the sample rehydration buffer is 0.5%.

*If you are loading 5-50 µg of protein (pure protein or crude lysate) per ZOOM strip, use 0.5% ampholytes in the sample rehydration buffer.
*If you are loading >50 µg of protein (crude lysate or fractionated sample) per ZOOM Strip, use 0.5-2% ampholytes in the sample rehydration buffer.

Note: Higher ampholyte concentration requires longer focusing times.

What is the maximum recommended volume of protein sample to be added to the Sample Rehydration buffer?

The maximum volume of the protein sample should at most be 1/6 of the final sample volume that will be added to the strip. A good general volume would be 5-10µL. 140 µL of sample diluted in Sample Rehydration buffer is used to rehydrate each ZOOM Strip for the standard rehydration time of one hour. For overnight rehydration, we recommend using 155 µL of diluted sample.

How should I store the ZOOM Carrier Ampholytes?

We recommend storing the ZOOM Carrier Ampholytes at 4 degrees C. Note: Storing some ZOOM Carrier Ampholytes at 4 degrees C may result in precipitate formation. Dissolve precipitate by warming the solution to 50 degrees C.

What are the storage conditions for the ZOOM Disks, ZOOM Carrier Ampholytes, ZOOM Focusing buffers, ZOOM Cathode buffer, IEF Anode buffer, and IEF Cathode buffer?

We recommend storing the ZOOM Disks, ZOOM Carrier Ampholytes, ZOOM Focusing buffers, ZOOM Cathode buffer, IEF Anode buffer, and IEF Cathode buffer at 4 degrees C.

What is the purpose of alkylation prior to IEF?

Alkylation prevents unwanted protein modifications by alkylating cysteines to avoid mixed disulfide formation and reoxidation and this allows for crisper focusing.

Do you still offer ZOOM 2D Protein Solubilizers?

Sorry we have discontinued selling the ZOOM 2D Protein Solubilizers.

What are ampholytes and do you offer them?

Ampholytes are molecules that contain both acidic and basic groups (and are therefore amphoteric) and will exist mostly as zwitterions in a certain range of pH. The pH at which the average charge is zero is known as the molecule's isoelectric point. Ampholytes are used to establish a stable pH gradient for use in isoelectric focusing. In gels containing ampholytes, a linear pH gradient is built up when an electric field is applied. The ampholyte molecules carry a net charge and thus migrate in the electric field between the electrodes as long as they reach the position of corresponding pI. They will then stop moving and form small plateaus (stationary stacks). Commercially available ampholytes are a mixture of synthetic molecules whose individual pI values cover a preselected pH range. Thus one can purchase carrier ampholytes spanning either a wide pH range (e.g., pH 3-10) or a narrow range (e.g., pH 7-8).

We offer ZOOM Carrier Ampholytes that are small, soluble molecules with both positive and negative charge groups. They sort at their relative positions based on their isoelectric points in an electric field, setting up a pH gradient. Carrier ampholytes help stabilize the pH gradient and current in IPG strips and aid in protein solubility, resulting in reproducible IEF resolution.

What are the main reasons for performing 2D gel electrophoresis of proteins?

The main advantages of performing 2D gel electrophoresis of proteins and applications used for are listed below:

Advantages:

*Simultaneous separation of hundreds to thousands of proteins
*High capacity with superior resolution
*Compatible with further analysis by MS for protein identification and sequencing
Ability to separate and analyze low-abundance proteins

Applications:

*Comparative proteomics: identifying and analyzing differences between complex mixtures of proteins
*Protein profiling, biomarker discovery
*Separation and analysis of protein variants and isoforms

What products do you offer for 2D gel electrophoresis of proteins?

We offer the following products for the first- and second-dimension separation of proteins:

First-dimension separation:

*Vertical gels for separation of proteins based on their isoelectric point (pI) (https://www.thermofisher.com/us/en/home/life-science/protein-expression-and-analysis/protein-gel-electrophoresis/protein-gels/specialized-protein-separation/isoelectric-focusing.html)

*Solution-phase isoelectric focusing of proteins using ZOOM Disks (immobilized buffer disks of specific pH) to reduce sample complexity, enrich low-abundance proteins, and increase the dynamic range of detection (https://www.thermofisher.com/us/en/home/life-science/protein-biology/protein-gel-electrophoresis/protein-gels/specialized-protein-gels/isoelectric-focusing/zoom-ief-fractionator.html)

*Mini gel system for high-throughput isoelectric focusing of proteins using ZOOM IPG (Immobilized pH Gradient) Strips (https://www.thermofisher.com/us/en/home/life-science/protein-biology/protein-gel-electrophoresis/protein-gels/specialized-protein-gels/2d-gel-electrophoresis/zoom-ipgrunner-system.html)

Second-dimension separation:

*ZOOM gels for 2D electrophoresis: NuPAGE Bis-Tris (Cat. No. NP0330BOX) and Tris-Glycine (Cat. No. EC60261BOX) mini gels with IPG wells ( to accommodate 7 cm ZOOM strips) for separation of proteins based on their molecular weight

What is 2D protein gel electrophoresis?

2D protein gel electrophoresis is the separation of proteins in two dimensions. In the first dimension, proteins are separated by their isoelectric point (pI) using isoelectric focusing, and in the second dimension, they are separated by their mass using SDS-PAGE.

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