Journal: Critical Care

Article Title: Type XVIII collagen degradation products in acute lung injury

doi: 10.1186/cc7779

Figure Lengend Snippet: Schematic representation of the human collagen XVIII variants, termed as SHORT, MIDDLE and LONG/FZ. Collagenous sequences are shown in white. Non-collagenous (NC) amino terminal sequences common to all variants are shown in black. Non-collagenous amino terminal sequences common to the two long variants are shown in grey. A non-collagenous amino terminal sequence specific to the LONG/FZ variant is marked with waves. The amino acid lengths (aa) and the predicted molecular masses (kDa) of these sequences are given, as are those of the full length type XVIII collagen molecules. Signal sequences are indicated by vertical and horizontal hatching. The epitopes of the antibodies used in this work are shown by black horizontal lines. The molecular masses of some endostatin-containing carboxy-terminal fragments are also indicated.

Article Snippet: A 25 μl sample of BALF was loaded onto polyacrylamide gel and the proteins were separated on a 7 to 12% SDS-PAGE under reducing conditions, electrotransferred to a nitrocellulose membrane (Protran; Schleicher & Schuell, Dassel, Germany) and probed with rabbit polyclonal antibodies against endostatin (HES.6) [ ] and human collagen XVIII (anti-all huXVIII QH4818 and anti-long huXVIII, QH1415.7) [ ], all used at a concentration 1 μg/ml in 5% fat-free milk powder in 1 × PBS, followed by a horseradish peroxidase-conjugated goat anti-rabbit antibody (Bio-Rad, Abingdon, UK).

Techniques: Sequencing, Variant Assay

Journal: Critical Care

Article Title: Type XVIII collagen degradation products in acute lung injury

doi: 10.1186/cc7779

Figure Lengend Snippet: Immunoprecipitation with anti-ALL antibody and detection with anti-LONG huXVIII antibody. Elevated type XVIII collagen precursors in the plasma of ALI patients compared with normal controls is shown. ALI = acute lung injury.

Article Snippet: A 25 μl sample of BALF was loaded onto polyacrylamide gel and the proteins were separated on a 7 to 12% SDS-PAGE under reducing conditions, electrotransferred to a nitrocellulose membrane (Protran; Schleicher & Schuell, Dassel, Germany) and probed with rabbit polyclonal antibodies against endostatin (HES.6) [ ] and human collagen XVIII (anti-all huXVIII QH4818 and anti-long huXVIII, QH1415.7) [ ], all used at a concentration 1 μg/ml in 5% fat-free milk powder in 1 × PBS, followed by a horseradish peroxidase-conjugated goat anti-rabbit antibody (Bio-Rad, Abingdon, UK).

Techniques: Immunoprecipitation, Clinical Proteomics

Journal: Critical Care

Article Title: Type XVIII collagen degradation products in acute lung injury

doi: 10.1186/cc7779

Figure Lengend Snippet: Western blot by anti-ALL huXVIII antibody. It shows elevated type XVIII collagen precursors in the bronchoalveolar lavage fluid (BALF) of patients with acute lung injury compared with normal controls. ALI = acute lung injury.

Article Snippet: A 25 μl sample of BALF was loaded onto polyacrylamide gel and the proteins were separated on a 7 to 12% SDS-PAGE under reducing conditions, electrotransferred to a nitrocellulose membrane (Protran; Schleicher & Schuell, Dassel, Germany) and probed with rabbit polyclonal antibodies against endostatin (HES.6) [ ] and human collagen XVIII (anti-all huXVIII QH4818 and anti-long huXVIII, QH1415.7) [ ], all used at a concentration 1 μg/ml in 5% fat-free milk powder in 1 × PBS, followed by a horseradish peroxidase-conjugated goat anti-rabbit antibody (Bio-Rad, Abingdon, UK).

Techniques: Western Blot