Anti-PKC gamma (phospho T655) antibody

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC gamma (phospho T655) antibody (AB5796)

Paraffin embedded human cerebellum tissue (right) stained for PKC gamma using ab5796 at 1/20 dilution in immunohistochemical analysis. Negative control without primary antibody (left).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC gamma (phospho T655) antibody (AB5796)

Paraffin embedded mouse cerebellum tissue (right) stained for PKC gamma using ab5796 at 1/100 dilution in immunohistochemical analysis. Negative control without primary antibody (left).

• WB

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Western blot - Anti-PKC gamma (phospho T655) antibody (AB5796)

Peptide Competition and Phosphatase treatment : Lysates prepared from HeLa cells stimulated with PMA were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. Membranes were either left untreated (1-11) or treated with lambda (ë) phosphatase (12), blocked with a 3% low-fat milk-TBST buffer overnight at 4°C, and incubated with 0.50 μg/mL ab5796 antibody for two hours at room temperature in a 3% BSA TBST buffer, following prior incubation with : no peptide (1, 11, 12), the non-phosphopeptide corresponding to the immunogen (2), a generic phosphothreonine containing peptide (3), the phosphopeptide immunogen (4), or, the phosphopeptide corresponding to the immunogen from other PKC isoforms (5-10). After washing, membranes were incubated with goat F(ab’)2 anti-rabbit IgG alkaline phosphatase and signals were detected using the Tropix WesternStarTM method. The data show that the peptide corresponding to PKC gamma [pT655] blocks the an

All lanes:

Western blot - Anti-PKC gamma (phospho T655) antibody (ab5796)

Predicted band size: 78 kDa

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• WB

CiteAb

Western blot - Anti-PKC gamma (phospho T655) antibody (AB5796)

Western Blotting using Anti-PKC gamma (phospho T655) antibody, ab5796. Publication image from Fujita, K. et al., 2018, Nat Commun, 29382817. Legend direct from paper.

PGRN inhibits downstream signals of Tyro3. a Western blot analyses show elevated phosphorylation of Tyro3 in the whole cerebral cortex sample prepared from PGRN-KI mice. Lower graphs show quantitation of pTyro3/GAPDH and pTyro3/Tyro3 ratios. *p < 0.05; **p < 0.01 (N = 6, Student’s t-test). Averages and s.e.m. are shown. b Western blot analyses show elevated phosphorylation of Shc was elevated in cerebral cortex of PGRN-KI mice. *p < 0.05; **p < 0.01 (N = 6, Student’s t-test). Averages and s.e.m. are shown. c Western blot analyses show elevated phosphorylation of PLCγ is increased in cerebral cortex of PGRN-KI mice. *p < 0.05; **p < 0.01 (N = 6, Student’s t-test). Averages and s.e.m. are shown. d Western blot analyses of Tyro3. Addition of Gas6 to culture medium increased phosphorylation of Tyro3 in cortical neurons (E15) at day 7 in primary culture, whereas co-addition of PGRN suppressed Gas6-induced phosphorylation of Tyro3. ##p < 0.01 (N = 4, Dunnett’s test). Averages and s.e.m. are shown. e Western blot analyses of Shc and B-Raf. Gas6 increased phosphorylation of Shc and B-Raf in mouse primary cortical neurons at day 7, whereas PGRN suppressed the activation induced by Gas6. ##p < 0.01 (N = 4, Dunnett’s test). Averages and s.e.m. are shown. f Western blot analyses of PLCγ and PKCα. Gas6 activated PLCγ and PKCα in mouse primary cortical neurons at day 7, whereas PGRN suppressed Gas6-induced activation of PKCα. PKCγ and PKCδ were not remarkably affected by Gas6. #p < 0.05; ##p<0.01 (N = 4, Dunnett’s test). Averages and s.e.m. are shown

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