goat polyclonal anti rat pedf antibody  (R&D Systems)

Journal: Cell Death & Disease

Article Title: Pigment epithelium-derived factor mediates retinal ganglion cell neuroprotection by suppression of caspase-2

doi: 10.1038/s41419-019-1379-6

Figure Lengend Snippet: a Fold change in CASP2 mRNA relative to β-actin reference gene in retinal cells freshly isolated and after 3d exposure to NBA, siCNL, 50 nM siCASP2 and PEDF/PEDF-34/PEDF-44. b % of surviving βIII-tubulin + RGC after 3d in culture in NBA, and after treatment with siCNL, 50 nM siCASP2 and PEDF/PEDF-34/PEDF-44. c Fold change in CASP2 mRNA relative to β-actin reference gene in retinal cells freshly isolated and after 3d exposure to suboptimal doses of siCASP2 (5–20 nM) with and without PEDF/PEDF-34/PEDF-44. d % of surviving βIII-tubulin + RGC after 3d in culture in with suboptimal concentrations of siCASP2 (5–20 nM) with and without PEDF/PEDF-34/PEDF-44 ( n = 9 wells/treatment)

Article Snippet: In a preliminary dose response experiment, the optimal concentration of goat polyclonal anti-rat PEDF antibody (Cat No. AF1177; R&D Systems Europe) to antagonise the effects of pre-optimised PEDF and PEDF-34 on RGC survival in vitro was determined , .

Techniques: Isolation

Journal: Cell Death & Disease

Article Title: Pigment epithelium-derived factor mediates retinal ganglion cell neuroprotection by suppression of caspase-2

doi: 10.1038/s41419-019-1379-6

Figure Lengend Snippet: a Fold change in CASP2 mRNA at 7d after ONC + PBS, ONC + siCNL and ONC + siCASP2 and ONC + PEDF treatment. b, c Western blot and subsequent densitometry to show that ONC-induced C-CASP2 (p12 fragment) was significantly suppressed by siCASP2 and PEDF treatment ( n = 18 retinae/treatment)

Article Snippet: In a preliminary dose response experiment, the optimal concentration of goat polyclonal anti-rat PEDF antibody (Cat No. AF1177; R&D Systems Europe) to antagonise the effects of pre-optimised PEDF and PEDF-34 on RGC survival in vitro was determined , .

Techniques: Western Blot

Journal: Cell Death & Disease

Article Title: Pigment epithelium-derived factor mediates retinal ganglion cell neuroprotection by suppression of caspase-2

doi: 10.1038/s41419-019-1379-6

Figure Lengend Snippet: a Fold change in CASP2 mRNA after ONC + saline and ONC + PEDF-34 daily eye-drops over 21d. b , c Western blot and subsequent densitometry to show that ONC-induced rise in C-CASP2 (p12 fragment) is significantly suppressed at 1d and 3d after injury and treatment, without changes in C-CASP3 or C-CASP6 levels. d Immunohistochemistry to show that ONC-induced localisation of C-CASP2 (red) in βIII-tubulin + (green) RGC (arrowheads) after ONC + saline treatment is suppressed by 1d and 3d after ONC + PEDF-34 eye-drops. Scale bar = 50 µm. GCL = ganglion cell layer. (n = 18 retinae/treatment)

Article Snippet: In a preliminary dose response experiment, the optimal concentration of goat polyclonal anti-rat PEDF antibody (Cat No. AF1177; R&D Systems Europe) to antagonise the effects of pre-optimised PEDF and PEDF-34 on RGC survival in vitro was determined , .

Techniques: Saline, Eye Drops, Western Blot, Immunohistochemistry

Journal: Cell Death & Disease

Article Title: Pigment epithelium-derived factor mediates retinal ganglion cell neuroprotection by suppression of caspase-2

doi: 10.1038/s41419-019-1379-6

Figure Lengend Snippet: a Fold change in CASP2 mRNA in 5-FDU-treated cultures showing no change in CASP2 mRNA after siCASP2 or PEDF treatment compared to cultures with glia. b % of surviving βIII-tubulin + RGC in 5-FDU treated cultures in siCASP2 treated cells was unaffected but survival was reduced in the absence of glia. c Fold change in CASP2 mRNA in purified RGC cultures showing no change in CASP2 mRNA after siCASP2 or PEDF treatment. d % of surviving βIII-tubulin + RGC in purified RGC cultures in siCASP2 treated cells was unaffected but survival was reduced in the absence of glia ( n = 9 wells/treatment)

Article Snippet: In a preliminary dose response experiment, the optimal concentration of goat polyclonal anti-rat PEDF antibody (Cat No. AF1177; R&D Systems Europe) to antagonise the effects of pre-optimised PEDF and PEDF-34 on RGC survival in vitro was determined , .

Techniques: Purification

Journal: Cell Death & Disease

Article Title: Pigment epithelium-derived factor mediates retinal ganglion cell neuroprotection by suppression of caspase-2

doi: 10.1038/s41419-019-1379-6

Figure Lengend Snippet: a ELISA to determine levels of BDNF, CNTF, GDNF, NGF and NT-3 released into the culture medium. Treatment of retinal cultures with ELISA isolated levels of BDNF, GDNF and NGF or CNTF either alone or in combination partially suppresses b CASP2 mRNA and C-CASP2 protein c , d by western blot and subsequent densitometry. e Representative images to demonstrate morphology of βIII-tubulin + RGC (note: neurite outgrowth occurs in surviving βIII-tubulin + RGC in wells containing PEDF, BDNF/GDNF/NGF and CNTF). (n = 9 wells/treatment). Scale bar in f = 20 µm

Article Snippet: In a preliminary dose response experiment, the optimal concentration of goat polyclonal anti-rat PEDF antibody (Cat No. AF1177; R&D Systems Europe) to antagonise the effects of pre-optimised PEDF and PEDF-34 on RGC survival in vitro was determined , .

Techniques: Enzyme-linked Immunosorbent Assay, Isolation, Western Blot

Journal: Cell Death & Disease

Article Title: Pigment epithelium-derived factor mediates retinal ganglion cell neuroprotection by suppression of caspase-2

doi: 10.1038/s41419-019-1379-6

Figure Lengend Snippet: a – c Pre-treatment of PEDF and PEDF-34 with a polyclonal antibody to PEDF fails to reduce CASP2 mRNA ( a ) and protein levels ( b ) whilst PEDF is suppressed in the absence of PEDF polyclonal antibody treatment, as confirmed by densitometry ( c ). d Pre-treatment with PEDF polyclonal antibody also suppresses the RGC neuroprotective effects of PEDF and PEDF-34. (n = 9 wells/treatment). e Representative images of the treatments in d to show morphology of βIII-tubulin + RGC. Scale bar in e = 20 µm

Article Snippet: In a preliminary dose response experiment, the optimal concentration of goat polyclonal anti-rat PEDF antibody (Cat No. AF1177; R&D Systems Europe) to antagonise the effects of pre-optimised PEDF and PEDF-34 on RGC survival in vitro was determined , .

Techniques:

Journal: Cell Death & Disease

Article Title: Pigment epithelium-derived factor mediates retinal ganglion cell neuroprotection by suppression of caspase-2

doi: 10.1038/s41419-019-1379-6

Figure Lengend Snippet: a CASP2-dependent apoptosis requires the formation of the PIDDosome (Adapted from ). b PEDF treatment suppresses CASP2, RAIDD but not PIDD and downstream Bim mRNA. c , d PEDF treatment suppresses C-CASP2, RAIDD, phopho-c-Jun and downstream Bim as detected by western blot and densitometry. e , f Immunohistochemistry to show changes in Bim protein colocalised to caspase-2 in the ganglion cell layer (GCL), inner plexiform layer (IPL), occasional cells of the inner nuclear layer (INL) whilst high power views of the GCL showed that Bim and caspase-2 were colocalised to RGC. Scale bars in e and ( f ) = 50 µm ( n = 18 retinae/treatment)

Article Snippet: In a preliminary dose response experiment, the optimal concentration of goat polyclonal anti-rat PEDF antibody (Cat No. AF1177; R&D Systems Europe) to antagonise the effects of pre-optimised PEDF and PEDF-34 on RGC survival in vitro was determined , .

Techniques: Western Blot, Immunohistochemistry