goat anti catx  (R&D Systems)

Journal: BMC Neuroscience

Article Title: Differential expression of Cathepsin S and X in the spinal cord of a rat neuropathic pain model

doi: 10.1186/1471-2202-9-80

Figure Lengend Snippet: CATS- and CATX-immunohistochemistry in normal rat spinal cord . Representative examples of CATS- (A, C, E-G) and CATX-immunostained (B, D, H-J) sections of the L5 segment. CATS-immunopositive deposits are localized in small glial-like cells (C, E, F) that distributed homogenously throughout the section (A), while CATX is mostly found in large neurons (D, H) and only few small cells are intensely stained (D, J). G, I: Sections incubated with preabsorbed primary antibodies are free of immunostaining. Scale bars, 500 μm (A, B), 50 μm (C, D), 20 μm (E-J).

Article Snippet: Proteins were electrophoretically separated on a 10% polyacrylamide gel containing SDS and transferred onto a PVDF-membrane (Carl Roth, Karlsruhe, Germany) at 4°C with 200 mA for 1.5 h. Blocking was performed with 1.5% milk powder and 1% BSA in TBS-Tween (0.1% Tween, 20 mM TBS) at RT for 1 h. Incubation with the primary antibodies goat anti-mouse CATX (1:500; R&D Systems, Wiesbaden, Germany) or goat anti-human CATS (1:200; R&D Systems) was conducted in blocking buffer overnight at 4°C.

Techniques: Immunohistochemistry, Staining, Incubation, Immunostaining

Journal: BMC Neuroscience

Article Title: Differential expression of Cathepsin S and X in the spinal cord of a rat neuropathic pain model

doi: 10.1186/1471-2202-9-80

Figure Lengend Snippet: Upregulation of CATS- and CATX-immunoreactivities in the spinal cord at 14 d after L5T . Survey micrographs illustrate the ipsilateral increase of CATS- and CATX-immunoreactivity in whole spinal cord sections (A, B), in the dorsal horn (DH) (C-F), the layer IX of the ventral horn (VH) and the fasciculus gracilis (FG). In the FG immunopositive cells exhibit macrophage-like morphology (K, M) and in the VH small immunopositive cells engulf motoneurons (G, I). At this time point the ipsilateral nucleus gracilis exhibits more intense CATS- (L) and CATX-staining (N) than the contralateral side. Scale bars, 500 μm (A, B, L, N), 20 μm (C-J), 10 μm (K, M).

Article Snippet: Proteins were electrophoretically separated on a 10% polyacrylamide gel containing SDS and transferred onto a PVDF-membrane (Carl Roth, Karlsruhe, Germany) at 4°C with 200 mA for 1.5 h. Blocking was performed with 1.5% milk powder and 1% BSA in TBS-Tween (0.1% Tween, 20 mM TBS) at RT for 1 h. Incubation with the primary antibodies goat anti-mouse CATX (1:500; R&D Systems, Wiesbaden, Germany) or goat anti-human CATS (1:200; R&D Systems) was conducted in blocking buffer overnight at 4°C.

Techniques: Staining

Journal: BMC Neuroscience

Article Title: Differential expression of Cathepsin S and X in the spinal cord of a rat neuropathic pain model

doi: 10.1186/1471-2202-9-80

Figure Lengend Snippet: Spatiotemporal progression of CATS-/CATX-immunoreactivities in the spinal cord after L5T . Cranial progression of cathepsin upregulation during the first 5 weeks after transection. The different expression patterns in the transverse plane are symbolized by different fillings of the bars. Both cathepsins exhibited the same spatial and temporal distribution pattern up to 35 d after transection.

Article Snippet: Proteins were electrophoretically separated on a 10% polyacrylamide gel containing SDS and transferred onto a PVDF-membrane (Carl Roth, Karlsruhe, Germany) at 4°C with 200 mA for 1.5 h. Blocking was performed with 1.5% milk powder and 1% BSA in TBS-Tween (0.1% Tween, 20 mM TBS) at RT for 1 h. Incubation with the primary antibodies goat anti-mouse CATX (1:500; R&D Systems, Wiesbaden, Germany) or goat anti-human CATS (1:200; R&D Systems) was conducted in blocking buffer overnight at 4°C.

Techniques: Expressing

Journal: BMC Neuroscience

Article Title: Differential expression of Cathepsin S and X in the spinal cord of a rat neuropathic pain model

doi: 10.1186/1471-2202-9-80

Figure Lengend Snippet: Upregulation of cathepsin protein levels and activities after L5T . A: Western blot analysis of CATX and CATS proform expression in the spinal cord of L5T (n = 5) and sham operated animals (n = 5) at 8 d after injury. Expression levels were normalized relative to the corresponding α-tubulin band. At this time point the L5 transection induced an upregulation of both proteins in all SC segments. The expression level of CATX was substantially higher than that of CATS. Data are means ± SD. B: CATX activities in the lumbar SC 8 d after transection in L5T, sham (n = 7, for each group) and naive animals (n = 4). Each symbol represents the value of a single animal, the bar indicates the mean for the group. CATX activity was significantly higher in L5T than in sham or naive SC. *** p ≤ 0.001. L, lumbar; C, cervical; T, thoracic.

Article Snippet: Proteins were electrophoretically separated on a 10% polyacrylamide gel containing SDS and transferred onto a PVDF-membrane (Carl Roth, Karlsruhe, Germany) at 4°C with 200 mA for 1.5 h. Blocking was performed with 1.5% milk powder and 1% BSA in TBS-Tween (0.1% Tween, 20 mM TBS) at RT for 1 h. Incubation with the primary antibodies goat anti-mouse CATX (1:500; R&D Systems, Wiesbaden, Germany) or goat anti-human CATS (1:200; R&D Systems) was conducted in blocking buffer overnight at 4°C.

Techniques: Western Blot, Expressing, Activity Assay

Journal: BMC Neuroscience

Article Title: Differential expression of Cathepsin S and X in the spinal cord of a rat neuropathic pain model

doi: 10.1186/1471-2202-9-80

Figure Lengend Snippet: Phenotyping of CATS and CATX cells . Double-immunfluorescence of the spinal cord shows colocalization of CATS with the astrocyte marker GFAP (A'-A"') and colocalization of CATX with the microglial marker PT66 (B'-B"') and the macrophage marker ED1 (C'-C"'). Large motoneurons expressed CATX (D) and CATS (E). Scale bars, 10 μm (A, B), 5 μm (C), 20 μm (D, E).

Article Snippet: Proteins were electrophoretically separated on a 10% polyacrylamide gel containing SDS and transferred onto a PVDF-membrane (Carl Roth, Karlsruhe, Germany) at 4°C with 200 mA for 1.5 h. Blocking was performed with 1.5% milk powder and 1% BSA in TBS-Tween (0.1% Tween, 20 mM TBS) at RT for 1 h. Incubation with the primary antibodies goat anti-mouse CATX (1:500; R&D Systems, Wiesbaden, Germany) or goat anti-human CATS (1:200; R&D Systems) was conducted in blocking buffer overnight at 4°C.

Techniques: Marker