WB
Western blot analysis using HSPA1L antibody against Hela(1),Hepg2(2),K562(3),A549(4),A431(5),Huvec(6),Nih3t3(7),Cos-7(8),C2C12(9),Hek293(10),Mcf-7(11),Pc-12(12),Jurkat(13),Raji(14) cell .
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 1:2000, 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 1:20000 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.
WB
Western blot analysis using HSPA1L antibody against Mouse skin tissue(1),Mouse kidney tissue(2),Mouse brain tissue(3),Mouse liver tissue(4),Mouse lung tissue(5),Mouse spleen tissue(6),Mouse stomach tissue(7),Mouse heart tissue(8),Rat brain tissue(9),Rat liver tissue(10),Rat lung tissue(11),Rat stomach tissue(12),Rat heart tissue(13) .
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 1:2000, 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 1:20000 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.
ICC/IF
Immunofluorescence analysis of C2C12 cells using HSPA1L 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.
ELISA
Coat the plate with immunizing antigen.Blue line: Immune serum; Orange line: Pre-immune serum.