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Testing Data
(Figure 1. Cell lysis protein yield with Cell Lysis Buffer Proteins were extracted from different cells following the Cell Lysis Buffer protocol. The protein concentration of each lysate was determined by BCA Protein Assay Kit to determine protein yield per milligram of starting cells.)
Mammal Cell Protein Extraction Reagent Western Blot Related Reagent
Mammal Cell Protein Extraction Reagent
Synonyms
Mammal Cell Protein Extraction Reagent; Mammal Cell Protein Extraction Reagent western blot related reagent
Form/Format
Liquid
Application Notes
Immunoassays: WB, ELISA, RIA, immunoprecipitation; reporter assays; protein kinase assays; protein purifications.
Compatible with additives, e.g. protease inhibitors (PMSF), reducing agents, chelating agents, salts
Compatible with additives, e.g. protease inhibitors (PMSF), reducing agents, chelating agents, salts
Enzymatic Activity
None; use additives to add function
Additional Materials Required
Procedure for Lysis of Monolayer-cultured Adherent Mammalian Cells
Note:Pre-chillan appropriatevolume of CellLysis Buffer at 4°C. If desired, add protease inhibitor and phosphatase inhibitor to the lysis bufferimmediately before use.
1. In a microcentrifuge tube, resuspend 5×106 cells in the growth media by scraping the cells off the surface of the plate with a cell scraper. Centrifuge harvested cell suspension at 600xg for 5min, then carefully remove and discard the supernatant.
2. Resuspend the cells in chilled PBS. Centrifuge at 600xg for 5min, then carefully remove and discard the supernatant.
3. Add 0.5 mL of chilled cell lysis buffer to the cell pellet. Vortex briefly. Incubate on ice for 30 minutes.
4. Centrifuge samples at10000xg for 10 minutes.
5. Transfer supernatant to a new tube for further analysis.
1. In a microcentrifuge tube, resuspend 5×106 cells in the growth media by scraping the cells off the surface of the plate with a cell scraper. Centrifuge harvested cell suspension at 600xg for 5min, then carefully remove and discard the supernatant.
2. Resuspend the cells in chilled PBS. Centrifuge at 600xg for 5min, then carefully remove and discard the supernatant.
3. Add 0.5 mL of chilled cell lysis buffer to the cell pellet. Vortex briefly. Incubate on ice for 30 minutes.
4. Centrifuge samples at10000xg for 10 minutes.
5. Transfer supernatant to a new tube for further analysis.
Procedure for Lysis of Suspension-cultured Mammalian Cells
Note: Pre-chillan appropriate volume of CellLysis Buffer at 4°C. If desired, add protease inhibitor and phosphatase inhibitor to the lysis buffer immediately before use.
1. Ina microcentrifuge tube, harvest 5×106 cells by centrifugation at 600xg for 5min. Carefully remove and discard the supernatant.
2. Resuspend the cells in chilled PBS. Centrifuge at 600xg for 5min, then carefully remove and discard the supernatant.
3. Add 0.5 mL of chilled Cell lysis buffer to the cell pellet. Vortex briefly. Incubate on ice for 30 minutes.
4. Centrifuge samples at 10000xg for 10 minutes.
5. Transfer supernatant to a new tube for further analysis.
1. Ina microcentrifuge tube, harvest 5×106 cells by centrifugation at 600xg for 5min. Carefully remove and discard the supernatant.
2. Resuspend the cells in chilled PBS. Centrifuge at 600xg for 5min, then carefully remove and discard the supernatant.
3. Add 0.5 mL of chilled Cell lysis buffer to the cell pellet. Vortex briefly. Incubate on ice for 30 minutes.
4. Centrifuge samples at 10000xg for 10 minutes.
5. Transfer supernatant to a new tube for further analysis.
Preparation and Storage
Shipped on blue ice. Upon receipt store at 4°C. Cell Lysis Buffer is stable for one year.
Testing Data
(Figure 1. Cell lysis protein yield with Cell Lysis Buffer Proteins were extracted from different cells following the Cell Lysis Buffer protocol. The protein concentration of each lysate was determined by BCA Protein Assay Kit to determine protein yield per milligram of starting cells.)
Testing Data
(Figure 1. Cell lysis protein yield with Cell Lysis Buffer Proteins were extracted from different cells following the Cell Lysis Buffer protocol. The protein concentration of each lysate was determined by BCA Protein Assay Kit to determine protein yield per milligram of starting cells.)
Testing Data
(Note: Cell lysis buffercan be added directly to the flask containing cells. Please see the following procedures.1. Carefully remove culture medium from adherent cells.2. Wash cells with chilled PBS. Carefully remove PBS.3. Add chilled Celllysis buffer to the cells. Vortex briefly. Incubate on ice for 30 minutes.(For the volume of the lysis buffer, follow the instructions listed below))
Testing Data
(Note: Cell lysis buffercan be added directly to the flask containing cells. Please see the following procedures.1. Carefully remove culture medium from adherent cells.2. Wash cells with chilled PBS. Carefully remove PBS.3. Add chilled Celllysis buffer to the cells. Vortex briefly. Incubate on ice for 30 minutes.(For the volume of the lysis buffer, follow the instructions listed below))
Testing Data
(Figure 2. Protein extraction from various cells using Cell Lysis Buffer Proteins were extracted from various cells following the Cell Lysis Buffer protocol. Lysates (20ug) were separated by SDS-PAGE and transferred to a nitrocellulose membrane. Incubate with primary antibodies. Images were generated using MyBioSource Hypersensitive ECL chemiluminiscence substrate.)
Testing Data
(Figure 2. Protein extraction from various cells using Cell Lysis Buffer Proteins were extracted from various cells following the Cell Lysis Buffer protocol. Lysates (20ug) were separated by SDS-PAGE and transferred to a nitrocellulose membrane. Incubate with primary antibodies. Images were generated using MyBioSource Hypersensitive ECL chemiluminiscence substrate.)
Testing Data
(Troubleshooting)
Testing Data
(Troubleshooting)
Related Product Information for Mammal Cell Protein Extraction Reagent western blot related reagent
Desciption: Cell Lysis Buffer is a ready-to-use Western blot related reagent solution used for efficient extraction of total soluble protein in nondenatured state from mammalian cells.
This Cell Lysis Buffer is a ready-to-use Western blot related reagent solution used forefficient extraction of total soluble protein in nondenatured state from mammalian cells. Cell lysis can be finished in 30 minutes. The cell lysis buffer has been validated for use with COS-7, NIH3T3, Hepa 1-6, 293T, CHO and other cell types.It is able to lyse plated cells and pelleted cells from suspension mammalian cultures.The produced cell lysates are directly compatible with reporter gene expression assays (luciferase, beta-galactosidase, chloramphenicol acetyl transferase, CAT, alkaline phosphatase), protein kinase assays (PKA, PKC, tyrosine kinase), phosphatase assays (general phosphatases, tyrosine phosphatases), immunoassays (Western blots, ELISAs, RIAs, immunoprecipitation), Coomassie-Blue and silver staining, BCAprotein assay, protein purification procedures, electrophoresis, and many other downstream applications
This Cell Lysis Buffer is a ready-to-use Western blot related reagent solution used forefficient extraction of total soluble protein in nondenatured state from mammalian cells. Cell lysis can be finished in 30 minutes. The cell lysis buffer has been validated for use with COS-7, NIH3T3, Hepa 1-6, 293T, CHO and other cell types.It is able to lyse plated cells and pelleted cells from suspension mammalian cultures.The produced cell lysates are directly compatible with reporter gene expression assays (luciferase, beta-galactosidase, chloramphenicol acetyl transferase, CAT, alkaline phosphatase), protein kinase assays (PKA, PKC, tyrosine kinase), phosphatase assays (general phosphatases, tyrosine phosphatases), immunoassays (Western blots, ELISAs, RIAs, immunoprecipitation), Coomassie-Blue and silver staining, BCAprotein assay, protein purification procedures, electrophoresis, and many other downstream applications
Product Categories/Family for Mammal Cell Protein Extraction Reagent western blot related reagent
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Product Notes
The Mammal Cell Protein Extraction Reagent (Catalog #AAA176736) is a Western Blot Related Reagent and is intended for research purposes only. The product is available for immediate purchase. Immunoassays: WB, ELISA, RIA, immunoprecipitation; reporter assays; protein kinase assays; protein purifications. Compatible with additives, e.g. protease inhibitors (PMSF), reducing agents, chelating agents, salts. Researchers should empirically determine the suitability of the Mammal Cell Protein Extraction Reagent for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "Mammal Cell Protein Extraction Reagent, Western Blot Related Reagent" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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