Thermo Scientific T-PER Tissue Protein Extraction Reagent enables mild extraction of total protein from tissue samples.
This tissue cell lysis reagent utilizes a proprietary detergent in 25mM bicine, 150mM sodium chloride (pH 7.6) to maximize the efficiency of protein solubilization from mammalian tissue samples by homogenization. The simple composition is compatible with additives such as protease inhibitors, salts, reducing agents, chelating agents, providing versatility for many different sample types and lysis applications. Cell lysates prepared with T-PER Reagent are directly compatible with reporter assays (e.g., luciferase, beta-galactosidase, chloramphenicol acetyl transferase), protein kinase assays (e.g., PKA, PKC, tyrosine kinase), immunoassays (e.g., Western blots, ELISAs, RIAs) and/or protein purification procedures.
- Simple procedure – just homogenize tissue sample in 1:20 (w/v) of tissue to T-PER Reagent, then centrifuge to pellet cell or tissue debris
- Mild and convenient – detergent is dialyzable for quick and easy removal; can be used with additional components (e.g., protease inhibitors, salts, reducing agents, chelating agents, etc.)
- Flexible – lysate may be used for reporter assays, protein kinase assays, immunoassays and/or protein purifications
- Compatible – the lysate is compatible with standard protein assays such as Pierce Coomassie Plus (Bradford) Protein Assay
Homogeneous and efficient tissue protein extraction from different samples is challenging due to varying tissue composition and extraction methods. Fibrous and collagenous tissues, like heart and kidney, make protein extraction by freeze-thaw or mortar and pestle methods inefficient without the aid of detergent. Conversely, soft tissues, such as spleen and liver, require a milder extraction protocol for optimal results. T-PER Reagent was designed for use with all tissues, independent of mechanical extraction methods. The mild detergent enhances solubilization of proteins without compromising function and the neutral pH and salt concentration of this reagent enable more efficient extraction from cellular compartments yielding homogeneous lysates.
T-PER Reagent was validated for protein yield in both fresh and frozen with frozen tissue and freeze-thaw extraction protocols predictably resulting in higher protein yield. Additionally, T-PER Reagent is shown to efficiently extract target proteins from various cellular compartments such as plasma membrane ( Na-K+ ATPase, pan-cadherin), cytoplasm (HSP90, MAPK), Mitochondria (COXIV), and nucleus (HDAC1) as tissue specific markers from brain (synaptophysin) and heart tissue (Troponin T Cardiac Isoform).
|T-PER Tissue Protein Extraction Reagent. Duplicate tissue samples were weighed, resuspended in 1:10 to 1:20 w/v T-PER Reagent and disrupted in a chilled Dounce or benchtop tissue homogenizer. The resulting lysates were centrifuged at 10,000 x g for 5 minutes and the supernatant was collected. The protein concentration of each lysate was determined using the Pierce BCA Protein Assay (Part No. 23227) to determine protein yield per milligram of starting tissue.
|Protein extraction from cellular compartments using T-PER Tissue Protein Extraction Reagent. Lysates from prepared using T-PER Reagent and extraction efficiency from the various cellular compartments of different tissues compared. For each target protein, 10µg of lysate was loaded for and electrophoresed by SDS PAGE, transferred to nitrocellulose membrane and detected by Western blot using SuperSignal West Pico Chemiluminescent Substrate (Part No. 34080).
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Review of cell lysis and protein extraction
Other Cell Lysis Reagents
Subcellular Protein Fractionation Kit for Tissues
N-PER Neuronal Protein Extraction Kit
Mitochondria Isolation Kit for Tissues
Protease and Phosphatase Inhibitor Cocktails and Tablets