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Next-Generation RIPA Buffer with Greatly Improved Membrane Protein Extraction Efficiency UltraRIPA Buffer

Date：October 06 2025Web Page No：160808

BioDynamics Laboratory Inc

UltraRIPA Buffer Appearance

UltraRIPA buffer is a next-generation RIPA buffer that can be used for experiments such as enzyme activity and immunoprecipitation because it can solubilize proteins under non-denaturing conditions, despite its higher extraction efficiency of membrane proteins than RIPA buffer. UltraRIPA buffer is useful for functional analysis of poorly soluble proteins such as membrane proteins on lipid rafts, which are difficult to solubilize with conventional non-denaturing cell solubilization buffers (RIPA buffer, 1% Triton X-100, etc.).

☞ Please also check out the UltraRIPA kit for Lipid Raft, a set of UltraRIPA Buffer and RIPA Buffer.

Features

  • The extraction efficiency of membrane proteins is higher than that of RIPA buffer (approximately 70% of proteins are solubilized from the RIPA-insoluble fraction).
    Membrane protein solubilization power: 2% SDS > UltraRIPA > RIPA > 1% Triton X-100
  • As with RIPA buffer, proteins can be extracted with little denaturation, so it can be used for analysis such as enzyme activity measurement.
    Protein denaturation: 2% SDS ≫ UltraRIPA ≈ RIPA ≈ 1% Triton X-100

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Example of Use

Comparison of Protein Solubilization Capacity

Comparison of Protein Solubilization Capacity_Method
Comparison of Protein Solubilization Capacity_Result

Experimental Procedures:
Whole mouse brains were mixed with RIPA buffer, disrupted by homogenizer and ultrasonic crusher, and centrifuged. After centrifugation, 2% SDS buffer, RIPA buffer, or UltraRIPA buffer was added to the precipitated fraction and resuspended. After centrifugation, protein content in each supernatant was compared by silver staining and BCA assay.

Experimental Results:
UltraRIPA buffer could extract more than 70% protein from the RIPA buffer-insoluble fraction.


Comparison of Extracted Enzyme Activities

Comparison of Extracted Enzyme Activities_Method
Comparison of Extracted Enzyme Activities_Results

Experimental Procedures:
CHO cells were lysed in 1% Triton X-100 buffer, RIPA buffer, UltraRIPA buffer, and 2% SDS buffer, respectively, and centrifuged. Lactate dehydrogenase (LDH) activity in the supernatants was measured.

Experimental Results:
LDH activity in the supernatants extracted with UltraRIPA buffer was comparable to that in the supernatants extracted with 1% Triton X-100 buffer or RIPA buffer.


Comparison of extractability from COS-1 cells

Protein solubilizing ability_method
Protein solubilizing ability_result

Experimental Procedures:
COS-1 cells were washed with PBS, lysed with SDS buffer, 1% Triton X-100 buffer, RIPA buffer and UltraRIPA buffer, and separated into soluble and insoluble fractions by centrifugation (14,000 rpm, 5 min, 4°C). The insoluble fraction was denatured and lysed with the same volume of SDS-PAGE sample buffer. The solubilized amount of flotilin 1, a lipid raft marker, was evaluated by SDS-PAGE/Western blot.

Each buffer:

  • SDS buffer (2% SDS, 1% Triton X-100, 50 mM HEPES-Na (pH 7.2), 150 mM NaCl)
  • RIPA buffer (1% NP-40 Alternative, 0.1% SDS, 50 mM Tris-HCl (pH8.0), 150 mM NaCl, 0.5% Sodium Deoxycholate)
  • UltraRIPA buffer (composition not disclosed)
  • 1% Triton X-100 buffer (1% Triton X-100, 50 mM HEPES-Na (pH 7.2), 150 mM NaCl)

Experimental Results:
In 1% Triton X-100 or RIPA buffer, most of Flotilin 1 remained in the insoluble membrane fraction, whereas in UltraRIPA buffer, almost all Flotilin 1 was dissolved.


Comparison of extractability from mouse brain tissue P2 membrane fraction

Comparison of extractability from mouse brain tissue P2 membrane fraction

Experimental Procedures:
P2 membrane fraction derived from mouse brain tissue (hippocampus + cerebral cortex) was prepared. 100 μl of each buffer was added to the P2 membrane fraction and sonicated on ice. The disrupted solution was centrifuged at high speed (100,000×g) to precipitate the insoluble fraction and collect each soluble fraction. 100 μl of 2% SDS buffer was added to each insoluble fraction for solubilization.

Each buffer:

  • SDS buffer (2% SDS, 50 mM Tris-HCl (pH 8.0), 150 mM NaCl)
  • RIPA buffer (1% NP-40 Alternative, 0.1% SDS, 50 mM Tris-HCl (pH8.0), 150 mM NaCl, 0.5% Sodium Deoxycholate)
  • UltraRIPA buffer (composition not disclosed)
  • 1% Triton X-100 buffer (1% Triton X-100, 50 mM Tris-HCl (pH 8.0), 150 mM NaCl)

Experimental Results:
UltraRIPA buffer showed higher solubility than 1% Triton X-100 or RIPA buffer for all neuronal synapse-related proteins tested.


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Product Information

[Date : October 08 2025 00:07]

Detail Product Name Product Code Supplier Size Price
UltraRIPA Buffer
DatasheetThis may not be the latest data sheet. 		F015B BDLBioDynamics Laboratory Inc 100 ml $220

Description Next-generation RIPA buffer that greatly improves the extraction efficiency of membrane proteins. Because proteins can be solubilized under non-denaturing conditions, it can also be used in experiments such as enzyme activity and immunoprecipitation.
Storage 4°C CAS
Link

UltraRIPA kit for Lipid Raft

[Date : October 08 2025 00:07]

UltraRIPA Buffer

DatasheetThis may not be the latest data sheet.


  • Product Code: F015B
  • Supplier: BDL
  • Size: 100ml
  • Price: $220

Description Next-generation RIPA buffer that greatly improves the extraction efficiency of membrane proteins. Because proteins can be solubilized under non-denaturing conditions, it can also be used in experiments such as enzyme activity and immunoprecipitation.
Storage 4°C CAS
Link

UltraRIPA kit for Lipid Raft

Go back to index

CONTACT

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