Protein in soluble form or not - (Jun/13/2006 )
I have extracted my protein with Tris, Nacl Buffer and Lysozyme and
Tris, Nacl, 0.1 % SDS and Lysozyme.
I have very sharp band with 1 % SDS . I juts load 20 ul of superntent.
Is my protein is forming inclusion body so that its sharp band with sds or no sharp band without sds. I am also trying with 8 M urea, 1 % 100 x Triton and 1% 114 X Triton.
Please suggest me from this picture that my protein is forming inclusion body or not.
after ladder first two rows are with Tris, NaCl, Lysozyme then Tris, NaCl, 0.1 % SDS and Lysozyme.
regards
-samita-
QUOTE (samita @ Jun 13 2006, 06:54 PM)
I have extracted my protein with Tris, Nacl Buffer and Lysozyme and
Tris, Nacl, 0.1 % SDS and Lysozyme.
I have very sharp band with 1 % SDS . I juts load 20 ul of superntent.
Is my protein is forming inclusion body so that its sharp band with sds or no sharp band without sds. I am also trying with 8 M urea, 1 % 100 x Triton and 1% 114 X Triton.
Please suggest me from this picture that my protein is forming inclusion body or not.
after ladder first two rows are with Tris, NaCl, Lysozyme then Tris, NaCl, 0.1 % SDS and Lysozyme.
regards
Tris, Nacl, 0.1 % SDS and Lysozyme.
I have very sharp band with 1 % SDS . I juts load 20 ul of superntent.
Is my protein is forming inclusion body so that its sharp band with sds or no sharp band without sds. I am also trying with 8 M urea, 1 % 100 x Triton and 1% 114 X Triton.
Please suggest me from this picture that my protein is forming inclusion body or not.
after ladder first two rows are with Tris, NaCl, Lysozyme then Tris, NaCl, 0.1 % SDS and Lysozyme.
regards
i would say it is maybe 30% soluble....you can try attach it to the column like that....did you try sonication or for example other detergents like sarcosyl before trying SDS and urea?
-Kathy-