RIPA Lysis Buffer
Referentie MBS2557000-50mL
Formaat : 50mL
Merk : MyBiosource
RIPA Lysis Buffer
For Research Use Only. Not for use in diagnostic procedures. Catalog #: MBS2557000Introduction: RIPA Lysis Buffer is a traditional rapid cell tissue lysate used as the preferred lysate for protein extraction from tissues or cells in the Western Blot assay. More details
Product Overview
Online Datasheet in PDFProduct Name
RIPA, Lysis Buffer
Full Product Name
RIPA Lysis Buffer
Form/Format
50 mM Tris (pH7.4), 150 mM NaCl, 1% Triton X-100, 1% C24H39O4Na, 1 mM EDTA, 0.1% SDS, 10 mM NaF, 1 mM Na3VO4, 1 mM PMSF
Preparation and Storage
Store at -20°C for 12 months
Instructions
1. For Tissue Samples
a. Take the samples, wash the tissue thoroughly with pre-cooled PBS (0.01M, pH7.4) to remove the surface blood and internal debris.
b. Weigh and smash the tissue, add an appropriate ratio of RIPA Lysis Buffer (add 10 uL PMSF and 10 uL Na3VO4 to 1 mL RIPA Lysis) and homogenizely lyse the tissue. It is recommended to homogenize according to the ratio of tissue weight: RIPA volume = 3:10. For example, add 1 mL RIPA Lysis Buffer to 0.3 g tissue sample, the specific volume can be adjusted according to experimental requirements.
c. Shake and lyse on the ice for 30 min after homogenization. Blow the sample repeatedly with a Pipette gun for about 50 times (under ice water bath conditions) to Make sure the DNA strand is broken and reduce the viscosity of sample.
d. Centrifuge at 12,000 rpm for 10 min at 4°C.
e. Take the supernatant and measure the protein protein concentration.
2. For Cell Sample
a. Collect the cells, wash them thoroughly with pre-cooled PBS (0.01 M, pH 7.4) to remove the medium off (it is generally recommended to wash 3 times).
b. Add an appropriate ratio of RIPA Lysis Buffer (10 uL PMSF and 10 uL Na3VO4 in 1 mL RIPA Lysis) and lyse on the ice for 30 min. It is recommended to add 0.1 mL of RIPA Lysis Buffer to each well of a 6-well plates (the protein content in different cells may vary, and the volume of the lysate added can be appropriately adjusted).
c. Blow the sample repeatedly with a Pipette gun for about 50 times (under ice water bath conditions) to make sure the DNA strand is broken and reduce the viscosity of the sample.
d. Centrifuge at 12,000 rpm for 10 min at 4°C.
e. Take the supernatant and measure the protein concentration.
a. Take the samples, wash the tissue thoroughly with pre-cooled PBS (0.01M, pH7.4) to remove the surface blood and internal debris.
b. Weigh and smash the tissue, add an appropriate ratio of RIPA Lysis Buffer (add 10 uL PMSF and 10 uL Na3VO4 to 1 mL RIPA Lysis) and homogenizely lyse the tissue. It is recommended to homogenize according to the ratio of tissue weight: RIPA volume = 3:10. For example, add 1 mL RIPA Lysis Buffer to 0.3 g tissue sample, the specific volume can be adjusted according to experimental requirements.
c. Shake and lyse on the ice for 30 min after homogenization. Blow the sample repeatedly with a Pipette gun for about 50 times (under ice water bath conditions) to Make sure the DNA strand is broken and reduce the viscosity of sample.
d. Centrifuge at 12,000 rpm for 10 min at 4°C.
e. Take the supernatant and measure the protein protein concentration.
2. For Cell Sample
a. Collect the cells, wash them thoroughly with pre-cooled PBS (0.01 M, pH 7.4) to remove the medium off (it is generally recommended to wash 3 times).
b. Add an appropriate ratio of RIPA Lysis Buffer (10 uL PMSF and 10 uL Na3VO4 in 1 mL RIPA Lysis) and lyse on the ice for 30 min. It is recommended to add 0.1 mL of RIPA Lysis Buffer to each well of a 6-well plates (the protein content in different cells may vary, and the volume of the lysate added can be appropriately adjusted).
c. Blow the sample repeatedly with a Pipette gun for about 50 times (under ice water bath conditions) to make sure the DNA strand is broken and reduce the viscosity of the sample.
d. Centrifuge at 12,000 rpm for 10 min at 4°C.
e. Take the supernatant and measure the protein concentration.
Cautions
1. All steps to lyse samples should be performed on ice or at 4°C.
2. It is recommended to add 10 uL PMSF and 10 uL Na3VO4 to 1 mL RAPI Lysis before use. If RIPA Lysis is precipitated, please dissolve at room temperature or in a warm water bath.
3. It is normal for a clean gelatinous substance to appear in the lysate of RIPA lysate. This transparent gel is a complex containing genomic DNA. Repeatedly blowing the sample with a pipette about 50 times can effectively reduce the viscosity of the sample. After centrifugation, the supernatant can be taken for subsequent experiments.
4. The tissue or cell sample lysed by RIPA cannot be measured by the Bradford method because of the high concentration of detergent in the lysis. It is recommended to measure the protein concentration by the BCA method.
5. For your safety and health, please wear the lab coat and disposable gloves before the experiments.
2. It is recommended to add 10 uL PMSF and 10 uL Na3VO4 to 1 mL RAPI Lysis before use. If RIPA Lysis is precipitated, please dissolve at room temperature or in a warm water bath.
3. It is normal for a clean gelatinous substance to appear in the lysate of RIPA lysate. This transparent gel is a complex containing genomic DNA. Repeatedly blowing the sample with a pipette about 50 times can effectively reduce the viscosity of the sample. After centrifugation, the supernatant can be taken for subsequent experiments.
4. The tissue or cell sample lysed by RIPA cannot be measured by the Bradford method because of the high concentration of detergent in the lysis. It is recommended to measure the protein concentration by the BCA method.
5. For your safety and health, please wear the lab coat and disposable gloves before the experiments.
ISO Certification
Manufactured in an ISO 9001:2015 Certified Laboratory.
Applications Tested/Suitable for RIPA lysis buffer
Western Blot (WB)
Other Notes
Small volumes of RIPA lysis buffer vial(s) may occasionally become entrapped in the seal of the product vial during shipment and storage. If necessary, briefly centrifuge the vial on a tabletop centrifuge to dislodge any liquid in the container`s cap. Certain products may require to ship with dry ice and additional dry ice fee may apply.
Searchable Terms for RIPA purchase
MBS2557000 is a lysis buffer targeting RIPA.
Additional Details
NCBI/Uniprot data below describe general gene information for RIPA. It may not necessarily be applicable to this product.
NCBI GI #
NCBI Official Full Name
RIPA
Protein Family
HTH-type transcriptional regulator
Precautions
All of MyBioSource's Products are for scientific laboratory research purposes and are not for diagnostic, therapeutics, prophylactic or in vivo use. Through your purchase, you expressly represent and warrant to MyBioSource that you will properly test and use any Products purchased from MyBioSource in accordance with industry standards. MyBioSource and its authorized distributors reserve the right to refuse to process any order where we reasonably believe that the intended use will fall outside of our acceptable guidelines.
Disclaimer
While every efforts were made to ensure the accuracy of the information provided in this datasheet, MyBioSource will not be liable for any omissions or errors contained herein. MyBioSource reserves the right to make changes to this datasheet at any time without prior notice.
It is the responsibility of the customer to report product performance issues to MyBioSource within 30 days of receipt of the product. Please visit our Terms & Conditions page for more information.
It is the responsibility of the customer to report product performance issues to MyBioSource within 30 days of receipt of the product. Please visit our Terms & Conditions page for more information.
Product Note
Select online data sheet information is drawn from bioinformatics databases, occasionally resulting in ambiguous or non-relevant product information. It is the responsibility of the customer to review, verify, and evaluate the information to make sure it matches their requirements before purchasing the kit. Our ELISA Kit assays are dynamic research tools and sometimes they may be updated and improved. If the format of this assay is important to you then please request the current manual or contact our technical support team with a presales inquiry before placing an order. We will confirm the current details of the assay. We cannot guarantee the sample manual posted online is the most current manual, it is intended to serve as an example only. Please refer to the instructions For Use provided with the assay kit for precise details.