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			96-Well Plate

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	48-602MAG	Buffer Detection Kit for Magnetic Beads	1 Kit

IP10 MilliporeProtein G Plus/Protein A-Agarose

A mixture of Protein G PLUS and Protein A covalently conjugated to agarose. Useful for purification of IgG from biological fluids.

Protein G Plus/Protein A-Agarose MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.

SDS
CoA
Data Sheet
Citations

Recommended Products

Overview

Replacement Information

Products

Catalogue Number	Packaging	Qty/Pack
IP10-10MLCN	Plastic ampoule	10 ml	Add To Favorites Add To Favorites Added To My Favorites

Description
Overview	Designed for immunoglobulin purification at low pressure. Product size refers to the volume of packed beads.
Catalogue Number	IP10
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). 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
IP10-10MLCN	04055977220247

Documentation

Protein G Plus/Protein A-Agarose SDS

Title
Safety Data Sheet (SDS)

Protein G Plus/Protein A-Agarose Certificates of Analysis

Title	Lot Number
IP10	Enter Lot Number

Citations

Title

Ellen J. Tisdale and Cristina R. Artalejo. (2006) Src-dependent a protein kinase C(aPKC) λ tyrosine phosphorylation is required for aPKCl/λ association with Rab2 and glyceraldehyde-3-phosphate dehydrogenase on pre-golgi intermediates. Journal of Biological Chemistry 281, 8436-8442.

Douglas M. Molina, Seema Grewal and Lee Bardwell. (2005) Characterization of an ERK-binding domain in Microphthalmia-associated transcription factor and differential inhibition of ERK2-mediated substrate phosphorylation. Journal of Biological Chemistry 280, 42051-42060.

Ellen J. Tisdale, Carmen Kelly and Cristina R. Artalejo. (2004) Glyceraldehyde-3-phosphate dehydrogenase interacts with Rab2 and plays an essential role in endoplasmic reticulum to golgi transport exclusive of its glycolytic activity. Journal of Biological Chemistry 279, 54046-54052.

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	04-September-2007 RFH
Application	Antibody Purification
Description	A mixture of Protein G PLUS and Protein A covalently conjugated to agarose. Useful for 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). To make Elution Buffer A, start with acetic acid and adjust the pH to 3.0 with ammonium hydroxide.
Storage	+2°C to +8°C
Do Not Freeze	Yes
Toxicity	Standard Handling
Citation	Ellen J. Tisdale and Cristina R. Artalejo. (2006) Src-dependent a protein kinase C(aPKC) λ tyrosine phosphorylation is required for aPKCl/λ association with Rab2 and glyceraldehyde-3-phosphate dehydrogenase on pre-golgi intermediates. Journal of Biological Chemistry 281, 8436-8442. Douglas M. Molina, Seema Grewal and Lee Bardwell. (2005) Characterization of an ERK-binding domain in Microphthalmia-associated transcription factor and differential inhibition of ERK2-mediated substrate phosphorylation. Journal of Biological Chemistry 280, 42051-42060. Ellen J. Tisdale, Carmen Kelly and Cristina R. Artalejo. (2004) Glyceraldehyde-3-phosphate dehydrogenase interacts with Rab2 and plays an essential role in endoplasmic reticulum to golgi transport exclusive of its glycolytic activity. Journal of Biological Chemistry 279, 54046-54052.

Catalogue Number	Description
IP02	Protein A Agarose Suspension
IP04	Protein G Plus-Agarose Suspension
IP05	Protein G Plus/Protein A Agarose Suspension

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