All sera included in this study were individually tested through the Outcherlony double immunodiffusion test using histoplasmin as an antigen (Guimarães et al., 2006 (link)). In addition, a Western blot assay (Pizzini et al., 1999 (link)) was performed to check the individual reactivity of the 30 included serum samples, as well as the three pool of sera described previously, against the protein extract prepared from the yeast cells of G217B H. capsulatum strain. Moreover, to check whether inespecific reactions could occur between the secondary antibody (alkaline phosphatase-conjugated AffiniPure goat anti-human IgG, Fc fragment specific, Jackson ImmunoResearch, West Grove, PA, USA) and the protein extract, a conjugate control was performed, where only the secondary antibody was added to the nitrocellulose strip with the antigen.
Affinipure goat anti human igg fc fragment specific
Manufactured by Jackson ImmunoResearch
Sourced in United States
AffiniPure Goat anti-human IgG Fc fragment specific is a laboratory reagent used to detect and quantify human immunoglobulin G (IgG) antibodies. It is an affinity-purified antibody that specifically binds to the Fc fragment of human IgG.
Automatically generated - may contain errors
Lab products found in correlation
96 well half area microplates, by Thermo Fisher Scientific (2 mentions)
Ma1 21315 1mg, by Thermo Fisher Scientific (2 mentions)
Alkaline phosphatase substrate pnpp tablets, by Merck Group (2 mentions)
Softmax version 5, by Molecular Devices (2 mentions)
Versamax microplate reader, by Molecular Devices (2 mentions)
3 protocols using affinipure goat anti human igg fc fragment specific
1
Immunoassay Validation for Histoplasma
2
ELISA for His-tagged Protein Detection
3
ELISA-based Protein Binding Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
ELISAs were performed on 96-well half-area microplates (ThermoFisher Scientific) as described previously15 (link). The plate was coated with 2 μg/mL mouse anti-His antibody (Invitrogen cat. #MA1-21315-1MG, ThermoFisher Scientific) overnight at 4°C. The following day, plates were washed three times with PBST (PBS + 0.05% Tween20) and incubated for 1h with blocking buffer (3% bovine serum albumin (BSA)). Following removal of blocking buffer, plates were treated with His-tagged proteins (5 μg/mL in PBST + 1% BSA) for 1.5h at room temperature. Plates were washed and serum was added at threefold dilutions (beginning at 1:30) and allowed to incubate for 1.5h. Following washes, secondary antibody (AffiniPure Goat anti-human IgG Fc fragment specific, Jackson ImmunoResearch Laboratories cat. #109-055-008) was added for an additional 1h. Secondary antibody was washed, and staining substrate (alkaline phosphatase substrate pNPP tablets, Sigma) was added. Absorbance at 405 nm was measured after 8, 20, and 30 min using VersaMax microplate reader (Molecular Devices) and analyzed using SoftMax version 5.4 (Molecular Devices).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!