Goat anti-Rabbit IgG (H&L), ALP conjugated
AS09 607 Clonality: Polyclonal Host: Goat Reactivity: Rabbit IgG (H&L)
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24 µg of Triticum aestivum L. whole leaf extract (1), 23 µg of Triticum aestivum L. whole leaf extract 37°С,3h (2), 22 µg of Triticum aestivum L. whole leaf extract, 37°С, 24h (3), 20 µg of Triticum aestivum L. whole leaf extract, 37°С,24h+50°C/1h (4), 17 µg of Triticum aestivum L. whole leaf extract, 37°С,24h+50°C, 3h (5), 23 µg of Triticum aestivum L. whole leaf extract, Control+50°C, 1h (6).
700 µg of total protein from spring wheat Triticum aestivum L. green leaves extracted with write exact buffer components 100 mM Tris HCl (pH=7.4), 1 mM beta-mercaptoethanol, 1 mM PMSF and denatured with 65.2 mM Tris HCl (pH=6.8), 1mM EDTA, 1% SDS, 20% glycerol, 5% beta-mercaptoethanol at 97°C for 5 min and 20 μg of total protein were separated on 12.5 % SDS-PAGE and blotted 2h on nitrocellulose membrane (GE Healthcare) using tank transfer. Blots were blocked with a skimmed milk 3 % in TBS for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1 000 for 1h at RT with agitation in TBS. The antibody solution was decanted and the blot washed 2 times for 10 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG, ALP conjugated, AS09 607, from Agrisera) diluted to 1:1000 in a skimmed milk 3 % in TBS for 1h at RT with agitation. The blot was washed 3 times for 5 min in TBS-T at RT with agitation and developed WB. The proteins were detected with 5-bromo-4-chloro-3-indolyl phosphate (Thermo Scientific) and Nitrotetrazolium Blue (Thermo Scientific). Exposure time was 5.20 minutes.
Courtesy of Dr. Olga Borovik, Laboratory of Physiological Genetics Siberian Institute of Plant Physiology and Biochemistry SB RAS Russia
Antibody has been affinity purified on solid phase rabbit IgG (H&L).
AP conjugate is supplied in 30 mM Triethanolamine, pH 7.2, 5 mM Magnesium Chloride, 0.1 mM Zinc Chloride, 1 % (w/v) BSA, Protease/IgG free. 0.05 % (w/v) of sodium azide is added as preservative
Based upon IEP, this antibody binds to:
heavy chains on rabbit IgG
light chains on all rabbit immunoglobulins
No reactivity is observed to non-immunoglobulin rabbit serum proteins based in immunoelectrophoresis.
Goat anti-rabbit IgG (H&L) is a secondary antibody conjugated to AP or ALP (Alkaline phosphatase) which binds to all rabbit immunoglobulins in immunological assays.
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