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Table 1 Recovery of Tat protein and removal of endotoxin

From: Transactivation and signaling functions of Tat are not correlated: biological and immunological characterization of HIV-1 subtype-C Tat protein

Tat Sample

Endotoxin (EU/μg)

Yield (mg/Liter)

 

B-Tat

C-Tat

B-Tat

C-Tat

Neat E. coli lysate

340

320

-

-

Ni-NTA (without Triton wash)

7.6

5.5

3.0

5.0

Ni-NTA (after Triton wash)

0.75

0.55

2.0

4.0

After SP-Sepharose

0.039

0.034

0.5

1.0

  1. Tat protein recombinantly expressed in bacteria was purified using Ni-NTA column chromatography and ion-exchange column chromatography performed sequentially. Protein concentration of the eluted Tat protein was determined using a dye binding assay [99]. Endotoxin concentration in the samples was determined using a commercial kit (QCL-1000, Biowhittaker).
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Retrovirology

ISSN: 1742-4690

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