One of the key factors influencing Western blot results is the extraction of proteins from cells. In practice, detergent-based buffers such as radioimmunoprecipitation assay (RIPA) buffer, along with physical disruption such as sonication, or the combination of both, have become the norm for protein extraction from cell membranes, cytoplasm, organelles, and nuclei.

Although RIPA buffer (with 0.1% SDS) or its substitute like NP-40 buffer (without SDS), has been widely used to lyse mammalian cells and tissues, RIPA buffer is not as effective in extracting large proteins as it is in medium and small proteins. To increase the harvest of large proteins, most laboratories combine RIPA buffer with sonication which can physically break down DNA to reduce the viscosity of the lysates. However, sonication has the potential to break down large proteins. Furthermore, to inhibit endogenous enzyme activities, inhibitors need to be added to the RIPA buffer.

IntactProtein™ Cell-Tissue Lysis Kit is formulated to solve these issues. It saves your time by avoiding the extra step of adding protease and phosphatase inhibitors; it can also preserve the structural and signal integrity of the cellular proteins. Further, this product is suitable for extracting all sizes of proteins from both adherent and suspension cells.

Key Features

• All-in-one formula: No protease/PTM inhibitors needed; no sonication required.

• Ready-to-use protocol: Simply mix Reagents A&B; extraction in as little as 15 min.

• Ultimate solution for large proteins: Near-complete extraction; no fragmentation.

• Assurance and peace of mind: No loss of protein PTMs such as phosphorylation.

• All-around performance: Suitable for mammalian cells and tissues.

Components

1. Reagent A: 40 μl
2. Reagent B: 20 ml

Storage

Reagent A at -20°C; Reagent B at room temperature.

Case Study

Publications

1. PPARγ Acetylation inAdipocytes Exacerbates BAT Whitening and Worsens Age-AssociatedMetabolic Dysfunction.
Publication: Cell Product: IntactProtein™ Cell-Tissue Lysis Kit

2. Salidroside reducesneuropathology in Alzheimer’s disease models by targeting NRF2/SIRT3pathway.
Publication: Cell Biosci Product: IntactProtein™ Cell-Tissue Lysis Kit

3. Ventromedial hypothalamic OGT drives adipose tissue lipolysis and curbs obesity.
Publication: Advanced Science Product: IntactProtein™ Cell-Tissue Lysis Kit

4. O-GlcNAcylation is a gatekeeper of porcine myogenesis.
Publication: J Anim Sci Product: IntactProtein™ Cell-Tissue Lysis Kit

5. PPARγAcetylation Orchestrates Adipose Plasticity and Metabolic Rhythms.
Publication: AdvSci (Weinh) Product: IntactProtein™ Cell-Tissue Lysis Kit

Customer Testimonials

“I was always having trouble dealing with extraction of large-sized proteins such as mTOR (MW 289) from cultured cells. My problem was that without sonication, the extraction efficiency of large proteins was low; with sonication, the large-sized proteins were fragmented into smaller-sized peptides. This dilemma was finally solved by using IntactProtein™ Cell-Tissue Lysis Kit. With this kit, I obtained excellent results without losing phosphorylation signals. Moreover, this kit was easy to use by simply mixing reagent A and B and lysing the cells for 15 min on ice.”

—— Virginia Tech, Dr. Xue Tian, Research Fellow

“We have tested your IntactProtein™ Cell-Tissue Lysis Kit in liver, adipose tissue, and cultured cells. It worked really well! Extracting proteins from fat is always a headache. Your buffer gave us high protein yields and sharp protein bands. We’d like to switch to your buffer”

—— Columbia University, Dr. Qiang Li, Assistant Professor