WB
Western blot analysis using Anti-IL27a-Mouse antibody against Recombinant IL27a-Mouse protein.
15% SDS-PAGE gel.Sample loading: 10ng /lane. Transfer the proteins onto a PVDF membrane (
OM790002), 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 1:5000, 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-Rabbit IgG(H&L)-HRP (
OM643487), at a ratio of 1:10000 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: 30S.
ELISA
Microtiter plate wells were coated with 100 ng/well coating antigen and incubated with serial dilutions of the antibody. Following HRP-conjugated secondary antibody and substrate development, OD450 was measured. The absorbance decreased progressively with increasing antibody dilution, yielding a sigmoidal titration curve that demonstrates the high antigen-binding activity of the polyclonal antibody.
ICC/IF
Immunofluorescence analysis of Hela cells using IL27a antibody (green). Blue: DAPI fluorescent DNA dye. Red: Actin filaments have been labeled with Omnimabs® 594-Phalloidin.Cells are fixed in 4% paraformaldehyde at room temperature for 15 minutes. Then, they are permeabilized with a PBS (OM750003) solution containing 0.1% Triton X-100(OM750021) at room temperature for 15 minutes. Subsequently, the cells are blocked with 10% non - immune goat serum(OM760028) at room temperature for 1 hour.The cells are incubated overnight at 4°C with the primary antibody diluted 1:100 in PBS. The secondary antibody, Omnimabs® 488 Goat Ant-Rabbit IgG(H&L) (Green,OM643486), is diluted at a ratio of 1:400 and incubated with the cells for 1 hour.Nuclear DNA is labeled with DAPI (Blue,OM643160). F-actin is stained with Omnimabs® 594-Phalloidin (Red,OM750007) diluted 1:100 for 30 minutes.