Address: bioBAY C10, 218 Xinghu Street, SIP, Suzhou, China
Postcode: 215123
Phone: 0512-69369088
Fax: 0512-69369089
website: www.abgent.com.cn
E-mail:[email protected]
[email protected]
Bacterial Protein Extraction Buffer
Catalog #: AC0462
Size: 500 mL
Description and Application:
Bacterial Protein extraction Buffer enables extraction of both soluble and insoluble
(inclusion bodies) proteins from bacteria.
The buffer provides high efficiency recovery of over-expressed proteins and is fully
compatible with HIS/GST-tag purification procedures and standard protein concentration
determination assays.
The buffer can also be used for protein extraction from insect cells.
Fomulated in: 20 mM Tris-HCl, pH 7.0 with a non-ionic detergent
Stability: >12 months at room temperature
Procedure (example for 250mL bacterial culture)
1. Pellet bacterial cells from a 250 mL culture at 5,000 g for 10 minutes.
Note: Extraction is typically more effective by freezing/thawing the cell pellet prior
proceeding to the next step.
2. Resuspend the pellet in 10 ml of extraction buffer and pipette up and down until the
suspension is homogenous.
Note: If desired, protease inhibitors can be added to the suspension.
3. Freshly prepare a 20 mg/ml stock solution of lysosyme in Bacterial Protein extraction
Buffer. Add 200 μl of the lysozyme stock solution to the cell suspension for a final
concentration of 0.4 mg/ml. Incubate on ice for 30 min with gentle shaking of the
suspension every 10 minutes.
Note: If sample becomes viscous, use DNaseI or sonicate the cell suspension (sonication
is recommended) until the suspension becomes fluid enough to ensure an efficient
centrifugation.
4. Centrifuge the suspension at 12,500g for 30 minutes and collect the supernatant.
OUR PRODUCTS ARE FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC OR THERAPEUTIC USE.