Home Antibody All anti-SERPINA1 antibodies
Also for SERPINA1 (NM_000295)
|a1-Anti-Trypsin [Human Plasma]|
||ELISA: 1:20,000 - 1:100,000, WB: 1:2,000 - 1:10,000, IHC: 1:500 - 1:2,500
|0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2|
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Goat-anti-Alpha-1-Anti-Trypsin (TA319139 lot 5842, red), Rabbit anti-Transferrin (109-4134 lot 3033), and Mouse-a-GST (200-301-200 lot 24882) were used in a multiplex system to detect target proteins under reducing (R) conditions (+4% BME) in albumin depleted human serum with 320 ng of added GST. Sample was run by SDS-PAGE, transferred to 0.2 um PVDF using the BioRad Trans-Blot Turbo and blocked in 2.5% Blotto, 2.5% BSA, 0.02% Tween over night at 4Â°C. Membrane was probed with three primary antibodies at 1:1000 dilution (in MB-070 over night at 4Â°C). Detection shown was using DyLight549 Donkey anti-Rabbit IgG (611-742-127 lot 21100, shown as green) Â DyLight 488 Donkey anti-Mouse IgG (610-741-124 lot 21095, shown as blue), Â and DyLight 649 Donkey anti-Goat IgG (605-743-125 lot 20834, shown as red) at 1:10000 (in MB-070 30 min RT). Blots were washed, rinsed in methanol, dried and Images were collected using the BioRad VersaDoc System.
Goat anti Alpha-1anti-Trypsin antibody (200-101-147 lot 5854) was used to detect Alpha-1anti-Trypsin under reducing (R) conditions. Reduced sample of purified target protein contained 4% BME and were boiled for 5 minutes. Samples of ~1ug of protein per lane were run by SDS-PAGE. Protein was transferred to nitrocellulose and probed with 1:3000 dilution of primary antibody (ON 4 C in MB-070). Detection shown was using Dylight 488 conjugated Donkey anti goat (605-741-125 lot 21094 1:10K in TBS/MB-070 1 hr RT). Images were collected using the BioRad VersaDoc System.
primary and Dylight conjugated secondary antibodies were used to detect: Human transferrin (1Â° 109-4134, green 2Â° 611-743-127); Alpha 1 anti trypsin (1Â° TA319139, red 2Â° 605-742-125); and Human IgG (1Â° 109-3102, Blue 2Â° 610-741-124 in a multiplex fluorescent western blot of human serum. Each primary antibody was diluted to 1:1000 in IRdye blocking buffer (MB-070) and incubated for 2 hrs at RT. Blot was 3X in TTBS, 1X in TBS and probed with secondary antibodies diluted 1:10000) in IRdye blocking buffer and incubated ~ 1hr at 4 degrees. After wash 2X in TTBS and 2X in TBS, blot was rinsed 2X in MeOH, dried and imaged using the Biorad VersaDoc4000.