Testing Data
(Figure 1. Tissue cell lysis protein yield with Mammalian Tissue Protein Extraction ReagentMouse tissue proteins were extracted from different mouse tissues following the Mammalian Tissue Protein Extraction Reagent protocol. The protein concentration of each lysate was determined by BCA Protein Assay Kit (Product No. MBS176737) to determine protein yield per milligram of starting tissue.)
Mammal Tissue Protein Extraction Reagent Western Blot Related Reagent
Mammal Tissue Protein Extraction Reagent
Applications
Western Blot, ELISA, Radioimmunoassay, Immunoprecipitation
Synonyms
Mammal Tissue Protein Extraction Reagent; Mammal Tissue Protein Extraction Reagent western blot related reagent
Form/Format
Liquid
Concentration
1mL of extraction reagent per 0.1g of tissue for tissue samples homogenization (varies by lot)
Applicable Applications for Mammal Tissue Protein Extraction Reagent western blot related reagent
Western Blot (WB), ELISA, RIA, Immunoprecipitation (IP); reporter assays; protein kinase assays; protein purifications; folding studies; chromatographic studies; DNA-protein interaction assays
Reagent Type
Universal tissue extraction buffer, detergent solution, cell lysis buffer
Usage
Obtaining homogenous tissue cell lysates with soluble protein extracts
Reactivity/severeness
Bland, minimal amount of detergent, detergent is dialyzable
Enzymatic Activity
None; use additives to add function
Additional components compatibility
Compatible with additives, e.g. protease inhibitors (PMSF), reducing agents, chelating agents,
Additional Materials Required
- Protease inhibitor (Product No. MBS176729) and phosphatase inhibitor (Product No. MBS176751)
- 2 ml microcentrifuge tubes
- Tissue homogenizer
- Microcentrifuge capable of spinning at 10,000 x g
- 2 ml microcentrifuge tubes
- Tissue homogenizer
- Microcentrifuge capable of spinning at 10,000 x g
Procedure for Tissue Lysis
Note: Pre-chill an appropriate volume of Mammalian Tissue Protein Extraction Reagent at 4°C. If desired, add protease inhibitor and phosphatase inhibitor to the lysis buffer immediately before use.
1. Place the fresh tissue into chilled PBS and rinse several times. Mince the tissue into small pieces.
2. Add Mammalian Tissue Protein Extraction Reagent to the tissue at 10:1. (i.e., Add 10 mL chilled lysis buffer per gram of tissue.) Use a smaller volume of reagent if a more concentrated protein extract is required.
3. Homogenize for several minutes at high speed until no tissue chunks remain.
4. Incubate on ice for 30 minutes.
5. Centrifuge at ~10000 x g for 10 minutes.
6. Transfer supernatant to a new tube for further analysis.
1. Place the fresh tissue into chilled PBS and rinse several times. Mince the tissue into small pieces.
2. Add Mammalian Tissue Protein Extraction Reagent to the tissue at 10:1. (i.e., Add 10 mL chilled lysis buffer per gram of tissue.) Use a smaller volume of reagent if a more concentrated protein extract is required.
3. Homogenize for several minutes at high speed until no tissue chunks remain.
4. Incubate on ice for 30 minutes.
5. Centrifuge at ~10000 x g for 10 minutes.
6. Transfer supernatant to a new tube for further analysis.
Precautions
- All steps of protein lysis should be operated on ice or at 4°C.
- Use BCA Protein Assay kit (Product No. MBS176737) to quantify lysed proteins. Bradford Protein Assay kit is not recommended.
- Use BCA Protein Assay kit (Product No. MBS176737) to quantify lysed proteins. Bradford Protein Assay kit is not recommended.
Preparation and Storage
Upon receipt store at 4°C. Mammalian Tissue Protein Extraction Reagent is stable for one year.
Testing Data
(Figure 1. Tissue cell lysis protein yield with Mammalian Tissue Protein Extraction ReagentMouse tissue proteins were extracted from different mouse tissues following the Mammalian Tissue Protein Extraction Reagent protocol. The protein concentration of each lysate was determined by BCA Protein Assay Kit (Product No. MBS176737) to determine protein yield per milligram of starting tissue.)
Testing Data
(Figure 1. Tissue cell lysis protein yield with Mammalian Tissue Protein Extraction ReagentMouse tissue proteins were extracted from different mouse tissues following the Mammalian Tissue Protein Extraction Reagent protocol. The protein concentration of each lysate was determined by BCA Protein Assay Kit (Product No. MBS176737) to determine protein yield per milligram of starting tissue.)
Testing Data
(Figure 2. Protein extraction from mouse tissues using Mammalian Tissue Protein Extraction Reagent Proteins were extracted from mouse tissues following the Mammalian Tissue Protein Extraction Reagent protocol. Lysates (20ug) were separated by SDS-PAGE and transferred to a nitrocellulose membrane. Incubate with primary antibodies. Images were generated using Hypersensitive ECL chemiluminiscence substrate (Product No. MBS177527).)
Testing Data
(Figure 2. Protein extraction from mouse tissues using Mammalian Tissue Protein Extraction Reagent Proteins were extracted from mouse tissues following the Mammalian Tissue Protein Extraction Reagent protocol. Lysates (20ug) were separated by SDS-PAGE and transferred to a nitrocellulose membrane. Incubate with primary antibodies. Images were generated using Hypersensitive ECL chemiluminiscence substrate (Product No. MBS177527).)
Related Product Information for Mammal Tissue Protein Extraction Reagent western blot related reagent
Mammalian Tissue Protein Extraction Reagent is a ready-to-use Western blot related reagent solution used for efficient extraction of total soluble protein in nondenatured state from animal tissues.
Description: Mammalian Tissue Protein Extraction Reagent is a ready-to-use Western blot related reagent solution used for efficient extraction of total soluble protein in nondenatured state from animal tissues by homogenization and cell lysis utilizing a dialyzable mild detergent at low concentration, for maximal protein solubilization and minimal interference with protein biological activity enabling direct use in downstream analysis.
Background: Extraction of tissue proteins being an essential step in the process of protein sample preparation is critical to successful protein analysis. Successful tissue protein extraction requires efficient cell lysis and protein solubilization, while preserving protein integrity and immunoreactivity. The varying tissue composition in different tissue samples implies different approaches for efficient and homogenous tissue protein extraction. The Mammalian Tissue Protein Extraction Reagent was designed for use with all tissues, independent of mechanical extraction methods. A mild nondenaturing and easily dialyzable detergent is added to the solution that aids in dissolving cell membranes and enhances solubilization of total protein without compromising function, and the neutral pH and salt concentration of the reagent enable more efficient extraction from cellular compartments yielding homogeneous lysates. 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, protein purification procedures, electrophoresis, folding studies, chromatographic studies, DNA-protein interaction assays (gel-shift assays), and many other downstream applications.
Description: Mammalian Tissue Protein Extraction Reagent is a ready-to-use Western blot related reagent solution used for efficient extraction of total soluble protein in nondenatured state from animal tissues by homogenization and cell lysis utilizing a dialyzable mild detergent at low concentration, for maximal protein solubilization and minimal interference with protein biological activity enabling direct use in downstream analysis.
Background: Extraction of tissue proteins being an essential step in the process of protein sample preparation is critical to successful protein analysis. Successful tissue protein extraction requires efficient cell lysis and protein solubilization, while preserving protein integrity and immunoreactivity. The varying tissue composition in different tissue samples implies different approaches for efficient and homogenous tissue protein extraction. The Mammalian Tissue Protein Extraction Reagent was designed for use with all tissues, independent of mechanical extraction methods. A mild nondenaturing and easily dialyzable detergent is added to the solution that aids in dissolving cell membranes and enhances solubilization of total protein without compromising function, and the neutral pH and salt concentration of the reagent enable more efficient extraction from cellular compartments yielding homogeneous lysates. 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, protein purification procedures, electrophoresis, folding studies, chromatographic studies, DNA-protein interaction assays (gel-shift assays), and many other downstream applications.
Product Categories/Family for Mammal Tissue Protein Extraction Reagent western blot related reagent
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Product Notes
The Mammal Tissue Protein Extraction Reagent (Catalog #AAA176732) is a Western Blot Related Reagent and is intended for research purposes only. The product is available for immediate purchase. AAA Biotech's Mammal Tissue Protein Extraction Reagent can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), ELISA, RIA, Immunoprecipitation (IP); reporter assays; protein kinase assays; protein purifications; folding studies; chromatographic studies; DNA-protein interaction assays. Researchers should empirically determine the suitability of the Mammal Tissue 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 Tissue 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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