Our broad portfolio consists of multiplex panels that allow you to choose, within the panel, analytes that best meet your needs. On a separate tab you can choose the premixed cytokine format or a single plex kit.
Cell Signaling Kits & MAPmates™
Choose fixed kits that allow you to explore entire pathways or processes. Or design your own kits by choosing single plex MAPmates™, following the provided guidelines.
The following MAPmates™ should not be plexed together:
-MAPmates™ that require a different assay buffer
-Phospho-specific and total MAPmate™ pairs, e.g. total GSK3β and GSK3β (Ser 9)
-PanTyr and site-specific MAPmates™, e.g. Phospho-EGF Receptor and phospho-STAT1 (Tyr701)
-More than 1 phospho-MAPmate™ for a single target (Akt, STAT3)
-GAPDH and β-Tubulin cannot be plexed with kits or MAPmates™ containing panTyr
.
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To begin designing your MILLIPLEX® MAP kit select a species, a panel type or kit of interest.
Custom Premix Selecting "Custom Premix" option means that all of the beads you have chosen will be premixed in manufacturing before the kit is sent to you.
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Add Additional Reagents (Buffer and Detection Kit is required for use with MAPmates)
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48-602MAG
Buffer Detection Kit for Magnetic Beads
1 Kit
Space Saver Option Customers purchasing multiple kits may choose to save storage space by eliminating the kit packaging and receiving their multiplex assay components in plastic bags for more compact storage.
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Designed for immunoglobulin purification at low pressure. Product size refers to volume of packed beads.
Catalogue Number
IP06
Brand Family
Calbiochem®
References
Product Information
Form
Liquid slurry
Formulation
50% suspension in PBS.
Preservative
≤0.1% sodium azide
Applications
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Application Notes
Antibody Purification
Application Comments
Note: the product size refers to the volume of packed beads. This product can be used directly with serum, plasma, tissue culture media, ascites, or other biological fluids, but if sufficient quantities of starting material are available we recommend an initial clean up step. The column life will be greatly extended if aggregated proteins and lipids are removed from the immunoglobulin in the clean up step. Use 5-10 ml of packed beads per ml serum.
Recommended Protocol for IgG Purification
Buffers
All concentrations stated are for working solutions, not the 10X concentrates. Caution: sodium azide is poison.
• Binding/Washing Buffer: 100 mM sodium phosphate pH 7.0, 150 mM sodium chloride, 5 mM sodium EDTA, 0.01% sodium azide. • Elution Buffer A (see Note section): 500 mM ammonium acetate pH 3.0, 0.01% sodium azide. • Elution Buffer B: 10 mM glycine/HCl pH 3.0, and 0.01% sodium azide. • Neutralization Buffer : 500 mM Tris Base, 0.01% sodium azide. • Storage Buffer: 100 mM sodium phosphate, pH 7.0, 0.01% sodium azide.
Protocol
A. Clean Up and Concentration Ascites and serum should be clotted at room temperature, refrigerated at 4°C overnight (to allow the clot to shrink and lipids to separate), and centrifuged multiple times to remove all clotted protein and lipid. Remove the lipid from the top of the centrifuge tube with a glass rod or small wooden stick. Tissue culture media should be centrifuged or filtered to remove aggregates. IgG can be concentrated and partially purified by use of an ammonium sulfate precipitation step. Add ammonium sulfate to 50% saturation (313 g per L) with stirring and check the pH adjusting to 7.0 by addition of 1 M HCl or NaOH. Centrifuge to collect precipitated immunoglobulin, dissolve in binding buffer and dialyze against the same buffer.
B. Purification 1. Pack a column with the Agarose Conjugate. 2. Wash with about 20 column volumes Washing/Binding Buffer until pH of eluate is 7.0. 3. If IgG has not been previously dialyzed against binding buffers dilute or dialyze IgG-containing sample into the Washing/Binding Buffer (pH 6.5-7.5). 4. Load sample onto column. 5. Wash with Washing/Binding Buffer until the absorbance of the eluate at 280 nm approaches background level. 6. Wash with Elution Buffer A to elute IgG, and collect fractions until A280 returns to background levels. 7. Wash with Elution Buffer B, and collect fractions until A280 returns to background. Most IgG should elute with buffer A. 8. Neutralize eluted IgG fractions by addition of an equal volume of neutralization buffer and check the pH with pH paper. For best results, neutralize eluate promptly. 9. To re-use the column immediately, repeat procedure from Step 2. 10. To prepare the column for storage, wash column with 5 column volumes of Elution Buffer B. 11. To store column wash with 30 column volumes storage buffer; then seal column outlets and store in refrigerator. 12. Quantitate the purified IgG using the formula:
Absorbance at 280 nm/1.4 = Concentration (mg/ml).
Note: To make Elution Buffer A, start with acetic acid and adjust the pH to 3.0 with ammonium hydroxide.
Biological Information
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Storage and Shipping Information
Ship Code
Blue Ice Only
Toxicity
Standard Handling
Storage
+2°C to +8°C
Do not freeze
Yes
Packaging Information
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Catalogue Number
GTIN
IP06
0
Documentation
Protein A Agarose Certificates of Analysis
Title
Lot Number
IP06
Data Sheet
Note that this data sheet is not lot-specific and is representative of the current specifications for this product. Please consult the vial label and the certificate of analysis for information on specific lots. Also note that shipping conditions may differ from storage conditions.
Revision
10-June-2010 RFH
Application
Antibody Purification
Description
Protein A covalently conjugated to agarose. Useful for the purification of IgG from biological fluids.
Background
The protein is covalently coupled to the agarose support and does not leach. The protein A will bind IgG from biological fluids, but with different efficiencies, depending on the species. In general, Protein A works well for purification of IgG of large mammals, while Protein G PLUS and a mixture of Protein G PLUS with Protein A are best for rodent IgG purifications.
Form
Liquid slurry
Formulation
50% suspension in PBS.
Preservative
≤0.1% sodium azide
Comments
Note: the product size refers to the volume of packed beads. This product can be used directly with serum, plasma, tissue culture media, ascites, or other biological fluids, but if sufficient quantities of starting material are available we recommend an initial clean up step. The column life will be greatly extended if aggregated proteins and lipids are removed from the immunoglobulin in the clean up step. Use 5-10 ml of packed beads per ml serum.
Recommended Protocol for IgG Purification
Buffers
All concentrations stated are for working solutions, not the 10X concentrates. Caution: sodium azide is poison.
• Binding/Washing Buffer: 100 mM sodium phosphate pH 7.0, 150 mM sodium chloride, 5 mM sodium EDTA, 0.01% sodium azide. • Elution Buffer A (see Note section): 500 mM ammonium acetate pH 3.0, 0.01% sodium azide. • Elution Buffer B: 10 mM glycine/HCl pH 3.0, and 0.01% sodium azide. • Neutralization Buffer : 500 mM Tris Base, 0.01% sodium azide. • Storage Buffer: 100 mM sodium phosphate, pH 7.0, 0.01% sodium azide.
Protocol
A. Clean Up and Concentration Ascites and serum should be clotted at room temperature, refrigerated at 4°C overnight (to allow the clot to shrink and lipids to separate), and centrifuged multiple times to remove all clotted protein and lipid. Remove the lipid from the top of the centrifuge tube with a glass rod or small wooden stick. Tissue culture media should be centrifuged or filtered to remove aggregates. IgG can be concentrated and partially purified by use of an ammonium sulfate precipitation step. Add ammonium sulfate to 50% saturation (313 g per L) with stirring and check the pH adjusting to 7.0 by addition of 1 M HCl or NaOH. Centrifuge to collect precipitated immunoglobulin, dissolve in binding buffer and dialyze against the same buffer.
B. Purification 1. Pack a column with the Agarose Conjugate. 2. Wash with about 20 column volumes Washing/Binding Buffer until pH of eluate is 7.0. 3. If IgG has not been previously dialyzed against binding buffers dilute or dialyze IgG-containing sample into the Washing/Binding Buffer (pH 6.5-7.5). 4. Load sample onto column. 5. Wash with Washing/Binding Buffer until the absorbance of the eluate at 280 nm approaches background level. 6. Wash with Elution Buffer A to elute IgG, and collect fractions until A280 returns to background levels. 7. Wash with Elution Buffer B, and collect fractions until A280 returns to background. Most IgG should elute with buffer A. 8. Neutralize eluted IgG fractions by addition of an equal volume of neutralization buffer and check the pH with pH paper. For best results, neutralize eluate promptly. 9. To re-use the column immediately, repeat procedure from Step 2. 10. To prepare the column for storage, wash column with 5 column volumes of Elution Buffer B. 11. To store column wash with 30 column volumes storage buffer; then seal column outlets and store in refrigerator. 12. Quantitate the purified IgG using the formula:
Absorbance at 280 nm/1.4 = Concentration (mg/ml).
Note: To make Elution Buffer A, start with acetic acid and adjust the pH to 3.0 with ammonium hydroxide.
