Tendon homogenization using the Minilys - Bertin Instruments

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Tendon homogenization using the Minilys

Sources: Proteomics Research in a Major Medical Center, Minnesota

Context

Proteomics involves the analysis and identification of protein structure and function. Through the Proteomics Core, investigators have access to a broad range of proteomics services to help them answer scientific questions. For this particular research project on tendon healing, the Minilys homogenizer was used to homogenize minced or intact tendon for protein extraction, followed by mass spectrometry analysis to identify unique peptides.

Results

Table 1. Protein extraction efficiency from dog tendon was compared between the CK68 and CKMix50 beads after homogenization using the Minilys.


Lane 1: Intact tendon, homogenized with CK68-R 2mL
Lanes 2 and 3: Intact (2) and minced (3) tendon, homogenized with CKMix50-R 2mL
Figure 1. Protein band patterns are compared using Coomassie Blue staining after SDS-PAGE electrophoresis.

The highest protein yields were observed by mincing the tendon first, prior to homogenization with the CKMix50-R 2mL tubes (Table 1). However, the protein band profiles of all three treatment groups were similar (Figure 1). Lane #3 was cut into different sections, trypsin-digested and analyzed using mass spectrometry, which led to the identification of 152 total unique proteins (a minimum of ≥2 peptides from each protein had to be detected for a confident ID). This included contaminating serum proteins like albumin, but the majority identified were unique collagen and extracellular matrix proteins.

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