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P3391

Millipore

Protein A-Sepharose® from Staphylococcus aureus

lyophilized powder

Synonym(s):
Protein A–Agarose
MDL number:
NACRES:
NA.56

Quality Level

form

lyophilized powder

packaging

glass bottle of 1 g
glass bottle of 1.5 g
glass bottle of 250 mg
glass bottle of 5 g

storage condition

do not freeze

extent of labeling

3 mg per mL

technique(s)

affinity chromatography: suitable

color

white to gray

matrix

Sepharose CL-4B

matrix activation

cyanogen bromide

matrix attachment

amino

matrix spacer

1 atom

capacity

≥16 mg/mL binding capacity (human IgG)

solubility

water: 1 g/L at 20 °C

suitability

conforms to structure for swelling capacity (1g swells to 4 to 5 mL)

storage temp.

2-8°C

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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.

Application

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.
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.

Packaging

1, 1.5, 5 g in glass bottle
250 mg in glass bottle

Quantity

Swelling: 1 g swells to approx. 4 ml.

Physical form

Supplied as lyophilized powder stabilized with lactose and dextran.

Legal Information

Sepharose is a registered trademark of Cytiva

Storage Class Code

11 - Combustible Solids

WGK

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Personal Protective Equipment

dust mask type N95 (US), Eyeshields, Gloves

Certificate of Analysis

Enter Lot Number to search for Certificate of Analysis (COA).

Certificate of Origin

Enter Lot Number to search for Certificate of Origin (COO).

Product Information Sheet

More Documents

Quotes and Ordering

  1. 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 (under Documents, above).

  2. 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).

  3. 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.

  4. 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.

  5. What is the binding capacity of Product P3391, Protein A-Sepharose®,  resin for my immunoglobulins?

    A table is available on the product information sheet (under Documents, above).

  6. 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.

  7. How do I get lot-specific information or a Certificate of Analysis?

    The lot specific COA document can be found by entering the lot number above under the "Documents" section.

  8. 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.

  9. 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. 

  10. My question is not addressed here, how can I contact Technical Service for assistance?

    Ask a Scientist here.

M Iezzi et al.
Molecular endocrinology (Baltimore, Md.), 13(2), 202-212 (1999-02-11)
Insulin-secreting cells express four GTPases of the Rab3 family. After separation of extracts of INS-1 cells on a sucrose density gradient, the bulk of the A, B, and C isoforms was recovered in the fractions enriched in insulin-containing secretory granules.
Lionel Gissot et al.
Plant physiology, 142(3), 931-944 (2006-10-10)
The sucrose nonfermenting-1 protein kinase (SNF1)/AMP-activated protein kinase subfamily plays a central role in metabolic responses to nutritional and environmental stresses. In yeast (Saccharomyces cerevisiae) and mammals, the beta- and gamma-noncatalytic subunits are implicated in substrate specificity and subcellular localization
T H Sisson et al.
Journal of immunological methods, 127(2), 215-220 (1990-03-09)
This report describes a modification of a procedure developed by others for crosslinking IgG to protein A which itself is covalently linked to a gel support. Earlier immunoaffinity columns were described as having large antigen-binding capacities and stability under a
Jyoti Parkash et al.
Endocrinology, 151(6), 2723-2735 (2010-04-08)
Within the preoptic region, nitric oxide (NO) production varies during the ovarian cycle and has the ability to impact hypothalamic reproductive function. One mechanism for the regulation of NO release mediated by estrogens during the estrous cycle includes physical association
Olga Khorosjutina et al.
The Journal of biological chemistry, 285(39), 29729-29737 (2010-07-14)
The multiprotein Mediator complex is an important regulator of RNA polymerase II-dependent genes in eukaryotic cells. In contrast to the situation in many other eukaryotes, the conserved Med15 protein is not a stable component of Mediator isolated from fission yeast.

Protocols

Immunoprecipitation Procedure

To determine the molecular weights of protein antigens, to study protein/protein interactions, to determine specific enzymatic activity, to monitor protein post-translational modifications and to determine the presence and quantity of proteins.

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

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