Non-modified FCS and Ca-FCS were coated at 10 µg/mL in 50 µL (diluted in pH 9.6 0.1M carbonate-bicarbonate buffer)(CB) on Nunc Maxisorp plates (Thermo Scientific) overnight. After washing in PBS containing 0.05% tween (Sigma)(PT), plates were blocked by incubating 100 µL PBS/1% bovine serum albumin (BSA)(Sigma) for 6 hours at 4°C. Following additional washing, wells were incubated with 50 µL serum at a 1/50 dilution in PBS/0.05% tween/1% BSA buffer (PTB) on ice overnight. All subsequent incubations were performed in PTB. As a standard, serial dilutions of a pool of positive sera were used. Human IgG was detected using rabbit anti-human IgG antibody (DAKO, Heverlee, Belgium) incubated on ice for 3.5 hours. After washing, wells were incubated on ice for 3.5 hours with horseradish peroxidase (HRP)-labeled goat anti-rabbit IgG antibody (DAKO). The final wash was followed by visualization of HRP enzyme activity using 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid)(ABTS) as substrate. We transformed the absorbance on both Ca-FCS and FCS to aU/mL and subtracted the background signal (aU/mL) of FCS from the signal (aU/mL) of CarP-FCS to analyze the specific anti-CarP-FCS reactivity.
Rabbit anti human igg antibody
Manufactured by Agilent Technologies
Sourced in Belgium, Denmark
The Rabbit anti-human IgG antibody is a laboratory reagent used for the detection and analysis of human immunoglobulin G (IgG) antibodies. It is a polyclonal antibody, raised in rabbits, that specifically binds to human IgG molecules.
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Lab products found in correlation
2 protocols using rabbit anti human igg antibody
1
Anti-Carbamylated Protein Antibody ELISA
2
IgG4-RD Submandibular Gland Analysis
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Specimens of submandibular glands for both IgG4-RD and controls and specimens of lymph nodes for controls were examined by immunohistochemical analysis of IgG, IgG4, IL-4, IL-13, IL-18 and IFN- expression. Three-µm-thick sections were prepared from each block of tissue embedded in paraffin. Deparaffinised sections were treated with 3% hydrogen peroxide for 10 min to inactivate endogenous peroxidase activity. The sections were incubated with protein blocker (Dako, Glostrup, Denmark) for 30 min and incubated at 4°C overnight with rabbit anti-human IgG antibody(1:5000, Dako), mouse anti-human IgG4 antibody (1:2000, Life technologies, Grand Island, NY), rabbit anti-human IL-18 antibody (PM014, 1:2000, Medical and Biological Laboratories CO., LTD., Nagoya, Japan), rabbit anti-human IL-13 antibody (06-1090, 1:200, MILLIPORE, MA, USA), rabbit anti-human IL-4 antibody (ab9622, 1:200, Abcam, Cambridge, UK) and rabbit anti human IFN- antibody (ab9657, 1:500, Abcam) as primary antibodies. The sections were washed three times with phosphate-buffered saline (PBS, pH 7.2). After washing with PBS, the sections were exposed to Envision + secondary antibody (Dako) for 30 min. The reaction products were developed by immersing the sections in a 3′3-diamidobenzidine tetrahydrochloride solution. The sections were counterstained with hematoxylin.
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