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P3391 Sigma

Protein A-Sepharose® from Staphylococcus aureus

lyophilized powder

Synonym: Protein A–Agarose



Related Categories Affinity Chromatography, Affinity Resins, Antibodies, Antibody Purification, Antibody Purification and Characterization,
form   lyophilized powder
extent of labeling   3 mg per mL
matrix   Sepharose CL-4B
matrix activation   cyanogen bromide
matrix attachment   amino
matrix spacer   1 atom
capacity   ≥16 mg/mL binding capacity (human IgG)
storage temp.   2-8°C


Frequently Asked Questions

Frequently Asked Questions are available for this Product.


1, 1.5, 5 g in glass bottle

250 mg in glass bottle


Swelling: 1 g swells to approx. 4 ml.

Physical form

Supplied as lyophilized powder stabilized with lactose and dextran.


Protein A-Sepharose was used:
• for immunoprecipitated placentas for protein and RNA analyses using Western blot technique.
• in chromatin immunoprecipitation to understand the link between Mediator and remodelling of chromatin structure at specific promoters.
• as lysis buffer to remove endogenous G-type Igs in protein extraction of brain samples from female rats.
• in immunoprecipitation of [32P] phosphomyristin C of rabbit antiserum, to study the down-regulation of protein kinase C (PKC) and phosphomyristin C (PMC) during T-cell development.

Protein A-Sepharose® is used for affinity chromatography, antibody purification and characterization, immunoaffinity matrices, protein chromatography, protein A, G and L resins, and recombinant protein expression and analysis. Protein A-Sepharose® has been used to develop strategies for investigating protein interactions, to improve the detection of celiac disease, and to study diabetes in children.

General description

Protein A-Sepharose when linked with antibody, helps improving antigen binding capacity of IgG crosslinked to column matrices. In this case, binding happens between Protein A and Fc portion of the IgG molecule leaving antigen specific sites free.

Legal Information

Sepharose is a registered trademark of GE Healthcare

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Safety & Documentation

Safety Information

NONH for all modes of transport
WGK Germany 

Frequently Asked Questions

How do I swell P3391, Protein A-Sepharose® resin?  
The lyophilized product should be swollen in buffer A for 30 minutes or longer at room temperature. Do not stir with any kind of mechanical stirrer.  One gram of powder typically swells to 3-4 mL of the hydrated gel.  This, along with other useful information (such as the formulation of buffer A) can be found on the product information sheet: http://www.sigmaaldrich.com/etc/medialib/docs/Sigma/Product_Information_Sheet/1/p3391pis.Par.0001.File.tmp/p3391pis.pdf
How can I store the hydrated P3391, Protein A-Sepharose® resin?  
Suspensions stored in the refrigerator in the presence of a suitable bacteriostatic agent, such as 20% ETOH, retain IgG binding activity. Another storage option is to put the resin into phosphate buffer saline, pH 7 with about 0.01% preservative (thimerosal or sodium azide).  This, along with other useful information can be found on the product information sheet: http://www.sigmaaldrich.com/etc/medialib/docs/Sigma/productinformationsheet2/p3391pis.Par.0001.File.tmp/p3391pis.pdf
What can P3391, Protein A-Sepharose® resin tolerate?  
Protein A Sepharose CL-4B exhibits high chemical and mechanical stability, and is stable over the pH range 2-11. It withstands high concentrations of denaturants, such as urea and guanidine hydrochloride. The gel displays high thermal stability but is not autoclavable. Furthermore, the gel can, if handled with care, be used many times.  This, along with other useful information, can be found on the product information sheet: http://www.sigmaaldrich.com/etc/medialib/docs/Sigma/productinformationsheet2/p3391pis.Par.0001.File.tmp/p3391pis.pdf
How do I clean Product P3391, Protein A-Sepharose®, resin?
It may be possible to clean the resin by washing it with 10-20 volumes of 100 mM Tris or borate buffer, pH8.5, containing 0.5-2.0 M NaCl, followed by 10-20 volumes of 100 mM acetate buffer, pH 4.0, containing0.5-2.0 M NaCl. Re equilibrate the resin with 20 volumes of buffer A.  The answer may also be found by clicking the link:   http://www.sigmaaldrich.com/sigma/product%20information%20sheet/p3391pis.pdf
What is the binding capacity of Product P3391, Protein A-Sepharose®,  resin for my immunoglobulins?
A table is available on the product information bulletin.  The answer may also be found by clicking the link:   http://www.sigmaaldrich.com/etc/medialib/docs/Sigma/Product_Information_Sheet/1/p3391pis.Par.0001.File.tmp/p3391pis.pdf  
Which document(s) contains shelf-life or expiration date information for a given product?
If available for a given product, the recommended re-test date or the expiration date can be found on the Certificate of Analysis. These documents are located on the product detail page under Useful Links & Tools. Click on the following link to search for a Certificate of Analysis. Please click the following link to see the details on our Product Dating Information.
How do I get lot-specific information or a Certificate of Analysis?
A Certificate of Analysis is available by lot number and can be obtained through our Advanced Search Option: http://www.sigmaaldrich.com/catalog/AdvancedSearchPage.do
How do I find price and availability?
There are several ways to find pricing and availability for our products.  Once you log onto our website, you will find the price and availability displayed on the product detail page. You can contact any of our Customer Sales and Service offices to receive a quote.  USA customers:  1-800-325-3010 or view local office numbers. 
What is the Department of Transportation shipping information for this product?
Transportation information can be found in Section 14 of the product's (M)SDS. To access the shipping information for this material, use the link on the product detail page for the product, or search here. 
My question is not addressed here, how can I contact Technical Service for assistance?
Use the option to the right to "Ask a Question" by email of a Technical Service Scientist.
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