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T-PER Tissue Protein Extraction Reagent


Mild solution designed for total protein extraction from tissue samples.

Thermo Scientific T-PER Tissue Protein Extraction Reagent

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.

Highlights:

  • 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

Product Details:

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).

Tissue cell lysis protein yield with T-PER Reagent
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.

 

Efficient tissue protein extraction from cellular compartments using T-PER Reagent
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).

References:

  1. Wright, MC., et al. (2014). Novel roles for osteopontin and clusterin in peripheral motor and sensory axon regeneration. J Neurosci., 34:1689-1700.
  2. Shareef, MM., et al. (2013). Interaction of HIF-1a and notch3 is required for the expression of carbonic anhydrase 9 in breast carcinoma cells. Genes & Cancer, 4:513-23.
  3. Tajima, S., et al. (2012). Iron reduction by deferoxamine leads to amelioration of adiposity via the regulation of oxidative stress and inflammation in obese and type 2 diabetes KKAy mice. Am J Physiol Endocrinol Metab., 302: E77-E86.
  4. Arthur, JC., et al. (2010). Cutting edge: NLRP12 controls dendritic and myeloid cell migration to affect contact hypersensitivity. J Immunol., 185:4515-19.
  5. Ikeda, Y. et al. et al. (2009). Cardiac-specific deletion of LKB1 leads to hypertrophy and dysfunction. J Biol Chem., 284:35839-49.
  6. Le, MTN., et al. et al. (2009). MicroRNA-125b is a novel negative regulator of p53. Genes & Dev., 23:862-76.
  7. Gupta, A., et al. (2009). Distinct functions of activated protein C differentially attenuate acute kidney Injury. J Am Soc Nephrol., 20:267-77.
  8. Chigurupati, S., et al. (2008). Lifelong running reduces oxidative stress and degenerative changes in the testes of mice. J Endocrinol., 199:333-41.
  9. Shyu, CY., et al. (2007). Subcellular transport of EKLF and switch-on of murine adult βmaj globin gene transcription. Mol Cell Biol., 27:2309-23.

Related Resources:

Review of cell lysis and protein extraction

Related Products:

Other Cell Lysis Reagents
Tissue Strainer
Subcellular Protein Fractionation Kit for Tissues
N-PER Neuronal Protein Extraction Kit
Mitochondria Isolation Kit for Tissues
Protease and Phosphatase Inhibitor Cocktails and Tablets


Ordering Information
 
Product # Description Pkg. Size Instructions MSDS CofA Price
78510 T-PER Tissue Protein Extraction Reagent
Formulation: Proprietary detergent in bicine-buffered saline, pH 7.6
Sufficient For: 25g fresh tissue
500mL Product Instructions for product #78510 T-PER Tissue Protein Extraction Reagent MSDS for product #78510 T-PER Tissue Protein Extraction Reagent $288.00

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Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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