Journal: The Journal of investigative dermatology

Article Title: Autoantibodies in Scurfy Mice and IPEX Patients Recognize Keratin 14

doi: 10.1038/jid.2010.16

Figure Lengend Snippet: (a) Mouse ear lysate was subjected to 2D gel electrophoresis, followed by Coomassie stain (left) or western blot analysis using a pool of five Scurfy sera as primary antibody (right). The numbers on the Coomassie-stained gel refer to spots identified by PMF as shown in Table 1. (b) Total keratin was extracted from freshly isolated keratinocytes from newborn mice, separated by SDS-PAGE, and analyzed by western blot using either a monoclonal mouse anti-human keratin 14 antibody (LL001), which is cross-reactive to mouse keratin 14, or a Scurfy serum as primary antibody.

Article Snippet: A vector with the complete mouse keratin 14 cDNA was purchased from OriGene (Austin, TX).

Techniques: Two-Dimensional Gel Electrophoresis, Electrophoresis, Staining, Western Blot, Isolation, SDS Page

Journal: The Journal of investigative dermatology

Article Title: Autoantibodies in Scurfy Mice and IPEX Patients Recognize Keratin 14

doi: 10.1038/jid.2010.16

Figure Lengend Snippet: Results of protein identification

Article Snippet: A vector with the complete mouse keratin 14 cDNA was purchased from OriGene (Austin, TX).

Techniques:

Journal: The Journal of investigative dermatology

Article Title: Autoantibodies in Scurfy Mice and IPEX Patients Recognize Keratin 14

doi: 10.1038/jid.2010.16

Figure Lengend Snippet: (a) Three protein fragments of keratin 14 were expressed and separated by SDS-PAGE, followed by Coomassie staining. (b) Fragments 1–3 were separated by SDS-PAGE, followed by western blot using a polyclonal rabbit anti-mouse keratin 14 antibody (AF64) or the corresponding isotype control as primary antibody. (c) The keratin 14 fragments were used either undiluted or at a 1:10 or 1:100 dilution for western blot with Scurfy serum as primary antibody. (d) The keratin 14 fragments were used for western blot as in (b), but with Scurfy, WT littermate, or nu/nu recipient serum as primary antibody. One representative experiment of at least three experiments with similar results is shown.

Article Snippet: A vector with the complete mouse keratin 14 cDNA was purchased from OriGene (Austin, TX).

Techniques: SDS Page, Staining, Western Blot

Journal: The Journal of investigative dermatology

Article Title: Autoantibodies in Scurfy Mice and IPEX Patients Recognize Keratin 14

doi: 10.1038/jid.2010.16

Figure Lengend Snippet: (a) Murine keratinocyte lysate was separated by SDS-PAGE, followed by western blot using sera of different immunodysregulation, polyendocrinopathy, enteropathy, X-linked (IPEX) patients or of a healthy donor. Serum from a Scurfy mouse served as positive control. One experiment of two with similar results is shown. (b) Human keratin lysate was separated by SDS-PAGE, followed by western blot using sera of different IPEX patients. The two right lanes show staining with either mouse anti-human keratin 14 antibody (LL001) or isotype control. One experiment of four with similar results is shown. (c) After preabsorption of the diluted IPEX sera on Escherichia coli proteins blotted onto polyvinylidene difluoride membranes, the sera were used separately for western blot analysis on the different fragments of murine keratin 14.

Article Snippet: A vector with the complete mouse keratin 14 cDNA was purchased from OriGene (Austin, TX).

Techniques: SDS Page, Western Blot, Positive Control, Staining