Recombinant human autoantigen Scl-70
A self-antigen, nucleic acid sequence technology, applied in the field of recombinant human self-antigen, can solve the problems of difficulty in extraction and purification of natural human antigens, limited development of ANA detection reagents, and difficulty in obtaining high-purity samples, and achieves strong reactivity, easy expression and The effect of purification and low cost of use
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Embodiment 1
[0032] Example 1: IPTG concentration is 0.2mM
[0033] The recombinant expression plasmid was introduced into Escherichia coli BL21 competent cells, and positive transformants were selected and inoculated in LB liquid medium containing kanamycin, and cultured overnight at 37°C on a shaking table. The next day, inoculate in 300ml of fresh same culture medium at a ratio of 1:100, culture on a shaker at 37°C until the A600 is 0.4-0.6, put the bacteria solution on ice for 30min, then add IPTG to a final concentration of 0.2mM, and overnight at 20°C induced expression.
Embodiment 2
[0034] Embodiment 2: IPTG concentration is 0.6mM
[0035] The recombinant expression plasmid was introduced into Escherichia coli BL21 competent cells, and positive transformants were selected and inoculated in LB liquid medium containing kanamycin, and cultured overnight at 37°C on a shaking table. The next day, inoculate 300ml of the same fresh culture medium at a ratio of 1:100, culture on a shaker at 37°C until the A600 is 0.4-0.6, put the bacteria in an ice bath for 30min, then add IPTG to a final concentration of 0.6mM, and overnight at 20°C induced expression.
Embodiment 3
[0036] Embodiment 3: IPTG concentration is 1.0mM
[0037] The recombinant expression plasmid was introduced into Escherichia coli BL21 competent cells, and positive transformants were selected and inoculated in LB liquid medium containing kanamycin, and cultured overnight at 37°C on a shaking table. The next day, inoculate 300ml of the same fresh culture medium at a ratio of 1:100, culture on a shaker at 37°C until the A600 is 0.4-0.6, put the bacteria in an ice bath for 30min, then add IPTG to a final concentration of 1.0mM, and overnight at 20°C induced expression.
[0038] The expression results induced by different IPTG concentrations are as follows: figure 1 shown. figure 1 Middle, M: Protein Marker, the size from top to bottom is 130KD, 95KD, 72KD, 55KD, 43KD, 34KD, 26KD, 17KD, 10KD, 1: blank control without adding IPTG, 2: adding 0.2mMIPTG, 3: adding 0.6mMIPTG, 4: add 1.0mMIPTG
[0039] from figure 1 It can be seen that after adding IPTG, the recombinant human auto...
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