WB
Western blot analysis using 14-3-3 eta antibody against Hela(1),A549(2),Mcf-7(3),Mouse liver tissue(4),Rat liver tissue(5),Mouse lung tissue(6),Rat lung tissue(7),Mouse brain tissue(8),Mouse kidney tissue(9),Mouse spleen tissue(10) lysate .
12% SDS-PAGE gel.Sample loading: 20μg /lane. Transfer the proteins onto a PVDF membrane (
OM790003), and block it with TBST (
OM750016) plus skimmed milk powder(OM750084) for one hour. Dilute the primary antibody with the antibody diluent (
OM750012) at a ratio of 1:1000, and incubate it overnight at 4°C. Wash the membrane three times with TBST (
OM750016), 5 minutes each time. At room temperature, dilute the secondary antibody, Goat Anti-Mouse IgG (H&L) - HRP(
OM644366), at a ratio of 1:20000 and incubate for one hour. Wash the membrane three times with TBST (
OM750016) again, 5 minutes each time. Use ECL (
OM625701) for luminescence.staining time: 60S
IHC
Immunohistochemical analysis of paraffin-embedded cervical cancer tissues using 14-3-3 eta antibody with DAB staining.Pre-treat the sections with heat-mediated antigen retrieval using sodium citrate buffer (pH 6.0) (OM750020) for 2 minutes. Wash the sections with ddH₂O and PBS (OM750003). Block the tissue with 10% non-immune goat serum(OM760028) at room temperature for 30 minutes. Incubate the tissue with the primary antibody diluted at a ratio of 1:500 at 4°C overnight. At room temperature, dilute the secondary antibody, Goat Anti-Mouse IgG(H&L)-HRP (OM644366), at a ratio of 1:200 and incubate for one hour. Use DAB(OM760029)as the chromogenic agent. Counterstain the tissue with hematoxylin, and mount the tissue sections with neutral gum.
IHC
Immunohistochemical analysis of paraffin-embedded muscle tissues using 14-3-3 eta antibody with DAB staining.Pre-treat the sections with heat-mediated antigen retrieval using sodium citrate buffer (pH 6.0) (OM750020) for 2 minutes. Wash the sections with ddH₂O and PBS (OM750003). Block the tissue with 10% non-immune goat serum(OM760028) at room temperature for 30 minutes. Incubate the tissue with the primary antibody diluted at a ratio of 1:500 at 4°C overnight. At room temperature, dilute the secondary antibody, Goat Anti-Mouse IgG(H&L)-HRP (OM644366), at a ratio of 1:200 and incubate for one hour. Use DAB(OM760029)as the chromogenic agent. Counterstain the tissue with hematoxylin, and mount the tissue sections with neutral gum.
IHC
Immunohistochemical analysis of paraffin-embedded prostate cancer tissues using 14-3-3 eta antibody with DAB staining.Pre-treat the sections with heat-mediated antigen retrieval using sodium citrate buffer (pH 6.0) (OM750020) for 2 minutes. Wash the sections with ddH₂O and PBS (OM750003). Block the tissue with 10% non-immune goat serum(OM760028) at room temperature for 30 minutes. Incubate the tissue with the primary antibody diluted at a ratio of 1:500 at 4°C overnight. At room temperature, dilute the secondary antibody, Goat Anti-Mouse IgG(H&L)-HRP (OM644366), at a ratio of 1:200 and incubate for one hour. Use DAB(OM760029)as the chromogenic agent. Counterstain the tissue with hematoxylin, and mount the tissue sections with neutral gum.
ELISA
Blue line: Antibody; Orange line: Isotype control antibody