Fractions were collected from each peak, after measuring the concentrations of their protein, these fractions were performed to SDS-PAGE and immunoblotting
for detecting the allergenic activity.
SDS-PAGE and immunoblotting
was performed using a mini gel system.
HCT-8 cells were stably transfected with human OAZ2 cDNA clone, and OAZ2 overexpression was verified by immunoblotting
One-dimensional SDS-PAGE and IgE and IgG1 immunoblotting
Then, the beads were washed four times with lysis buffer, and the bound proteins were analyzed by immunoblotting
using the indicated antibodies.
Anti-Smith (Sm) antibody, which was first described by Tan and Kunkel,[sup] is specific for adult systemic lupus erythematosus (SLE) according to the American College of Rheumatology.[sup] However, the frequency of anti-Sm antibody in adult SLE patients is low (ranging from 5% to 30%) depending on the serologic tests used and the ethnic origin of the patient population.[sup], More researchers have exploited various techniques, such as immunoblotting
, immunofluorescence, and enzyme-linked immunosorbent assay (ELISA), to increase anti-Sm antibody detection sensitivity.[sup],,,,
To identify the allergen(s) in the cyanobacteria that is binding to IgE, we performed 2D gel electrophoresis, followed by immunoblotting
, using MC(-) lysate and pooled patient sera.
findings were largely divided into three patterns (Figure 1): pattern 1: nonglycosylated form (NG) dominant (corresponding to percentage of glycosylation <40% in Figure 3), pattern 3: glycosylated form (G) dominant (corresponding to percentage of glycosylation >60% in Figure 3), and pattern 2: intermediate between patterns 1 and 3.
. The WCA and SSA of all the isolates separated on 12.5% (w/v) SDS-PAGE slabs  were transferred electrophoretically on nitrocellulose membrane papers (NCP) (Sartorius).
The results of immunoblotting
inhibition revealed that the IgE binding reactivity of the allergenic proteins with 12, 20, 39, and 45 from the A.
revealed that DAT substrates led to increased DAT T53 phosphorylation.
To collect urine, both ureters were exposed and catheterized through a midline incision as described by Topcu and colleagues.[sup.15] Before sacrificing the animals, we drew about 2 mL of artery blood from the abdominal aorta in gas-tight syringes; both kidneys were then quickly removed for immunoblotting
. The blood was centrifuged for 15 minutes at 4000 g to remove the blood cells, and the plasma was subsequently analyzed for sodium, potassium, creatinine and urea (Hitachi 7600-020 automatic biochemical analyzer, Hitachi High-Technologies, Tokyo, Japan).