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.
P3391
Protein A-Sepharose™ from Staphylococcus aureus
lyophilized powder
Synonym(s):
Protein A–Agarose
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About This Item
Form:
lyophilized powder
Pricing and availability is not currently available.
form
lyophilized powder
Quality Segment
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
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.[1]
Application
Protein A-Sepharose was used:
- for immunoprecipitated placentas for protein and RNA analyses using Western blot technique.[2]
- in chromatin immunoprecipitation to understand the link between Mediator and remodelling of chromatin structure at specific promoters.[3]
- as lysis buffer to remove endogenous G-type Igs in protein extraction of brain samples from female rats.[4]
- 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.[5]
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.
Physical form
Supplied as lyophilized powder stabilized with lactose and dextran.
Preparation Note
Swelling: 1 g swells to approx. 4 ml.
Legal Information
Sepharose is a trademark of Cytiva
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This Item | |||
|---|---|---|---|
| technique(s) affinity chromatography: suitable | technique(s) affinity chromatography: suitable | technique(s) ELISA: suitable, immunohistochemistry: suitable, immunoprecipitation (IP): suitable, western blot: suitable | technique(s) ELISA: suitable, immunohistochemistry: suitable, immunoprecipitation (IP): suitable, western blot: suitable |
| suitability conforms to structure for swelling capacity (1g swells to 4 to 5 mL) | suitability - | suitability - | suitability - |
| form lyophilized powder | form aqueous ethanol suspension | form powder | form essentially salt-free, lyophilized powder |
| storage temp. 2-8°C | storage temp. 2-8°C | storage temp. 2-8°C | storage temp. 2-8°C |
| storage condition do not freeze | storage condition - | storage condition - | storage condition - |
| solubility water: 1 g/L at 20 °C | solubility - | solubility H2O: 1 mg/mL, clear, colorless | solubility - |
Storage Class
11 - Combustible Solids
wgk
WGK 3
flash_point_f
Not applicable
flash_point_c
Not applicable
ppe
Eyeshields, Gloves, type N95 (US)
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What can P3391, Protein A-Sepharose® resin tolerate?
1 answer-
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How do I swell P3391, Protein A-Sepharose® resin?
1 answer-
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).
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What is the binding capacity of Product P3391, Protein A-Sepharose®, resin for my immunoglobulins?
1 answer-
A table is available on the product information sheet (under Documents, above).
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What is the Department of Transportation shipping information for this product?
1 answer-
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.
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How can I store the hydrated P3391, Protein A-Sepharose® resin?
1 answer-
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).
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How do I clean Product P3391, Protein A-Sepharose®, resin?
1 answer-
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.
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