Serum components binding to AdV ELISAs; goat anti-human IgM HRP, goat anti-human IgA HRP, goat anti-human IgG HRP, mouse anti-human IgD HRP, and mouse anti-human IgE HRP were obtained from Abcam. Detection of human C3b, C4b, Mannan-binding lectin, and C-reactive protein were modified from ELISA detection kits from Abcam. Pentraxin 3 detection was by modified ELISA kits from Hycult Biotech. Immunoblot of CD11b, CD11c, CD18, CD35, CD46, CD55, p62 and Complement C3 was by antibodies from Abcam. Immunoblot of RIG-I, MDA5, phosphorylated IKK, total IKK, phosphorylated IκB, total IκB, phopshorylated NF-κB p65, total p65, TRAF2, TRAF3, TRAF5, TRAF6 and GAPDH (loading control) used antibodies from Cell Signaling Technologies. Immunoblot for STING and MAVS used antibodies from Santa Cruz Biotechnology. Immunoblot and immunofluorescence for AdV hexon used polyclonal goat anti-adenovirus 5 antibody from Millipore. Immunofluoresence used MAVS and TOM20 antibodies from Santa Cruz.
Goat anti human igg hrp
Manufactured by Abcam
Sourced in United States
Goat anti-human IgG-HRP is a secondary antibody conjugated with horseradish peroxidase (HRP). It is designed to detect and measure human immunoglobulin G (IgG) in various immunoassays and experiments.
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Lab products found in correlation
Mouse anti human ige hrp, by Abcam (1 mentions)
Elisa detection kits, by Abcam (1 mentions)
3 3 diaminobenzidine (dab), by Tiangen Biotech (1 mentions)
Elisa microplate reader, by Agilent Technologies (1 mentions)
Sulfuric acid, by Thermo Fisher Scientific (1 mentions)
Ultra tmb, by Thermo Fisher Scientific (1 mentions)
Blocker bsa, by Thermo Fisher Scientific (1 mentions)
Xmark spectrophotometer, by Bio-Rad (1 mentions)
Sars cov 2 surrogate virus neutralization test, by GenScript (1 mentions)
Human igg isotype control, by Abcam (1 mentions)
1 step ultra tmb elisa substrate solution, by Thermo Fisher Scientific (1 mentions)
96 well flat bottom plate, by Corning (1 mentions)
Superblock blocking buffer, by Thermo Fisher Scientific (1 mentions)
Polyvinylidene fluoride pvdf membrane, by Merck Group (1 mentions)
Oligonucleotide primers, by Genewiz (1 mentions)
8 protocols using goat anti human igg hrp
1
Multiplexed AdV Serum Profiling
2
EphA2 Immunohistochemical Staining Protocol
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Immunohistochemistry was performed as previously described [58 (link)]. Adenocarcinoma and adjacent normal lung tissues were fixed, dehydrated, paraffin-embedded, and sectioned. The dewaxed sections were placed in hydrogen peroxide (containing 3% methanol) for 10 min at room temperature and washed with 1 × PBS. The tissue sections were immersed in 0.01 M citrate buffer solution (pH 6.0) and heated to boiling. Following cooling, the sections were washed with 1 × PBS. Blocking solution (goat serum) was added dropwise to block for 20 min at room temperature. The purified bivalent recombinant antibody, EphA2-scFv-Fc, and the full-length antibody, EphA2-IgG1 was separately added dropwise as a primary antibody. Moreover, the human IgG1 (Shanghaiyuanye Biotechnology, Shanghai, China) was used as an isotypic control and incubated overnight at 4°C. Goat-anti-human-IgG-HRP (Abcam, Cambridge, USA) was added dropwise as a secondary antibody and incubated for 90 min at 37°C and washed with 1 × PBS at the end of the incubation. A DAB chromogenic reagent kit (Zhongshan Jinqiao, Beijing, China) was used to stain at room temperature. After hematoxylin counterstaining and dehydration, the sections were sealed with neutral gum.
3
Immunoblotting of Recombinant CD40 Proteins
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His-tagged recombinant CD40 proteins were run on 18% polyacrylamide gels and transferred onto nitrocellulose membranes. The membranes were blocked in PBS 5% non-fat dried milk 0.05% Tween 20 and then incubated with various anti-CD40 mAb at 4°C overnight before detection using secondary polyclonal goat anti-human IgG-HRP (Abcam, UK). The membranes were washed with PBS before the addition of ECL and the signals were captured on a UVP Biospectrum Imaging System.
4
Indirect ELISA for SLE Autoantibody Detection
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Serum samples were collected from all SLE patients and HCs. 100 µl EP (10 µg/mL) was coated in each microplate. 1:50 diluted serum sample was used as primary antibody and sealed with 5% skim milk, while 1:10,000 diluent goat anti-human IgG-HRP (Abcam, USA) was applied as secondary antibody to carry out indirect ELISA. Post being incubated with DAB (TIANGEN, China), OD was subsequently measured at 450 nm by using a Bio-Tek ELISA microplate reader. All samples were independently analyzed in triplicate, and the mean value of OD was used for analysis in this study.
5
SARS-CoV-2 Antibody Quantification
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Total SARS-CoV-2 Spike-binding or RBD:ACE2 inhibiting antibody levels were quantified in plasma using ELISA kits: SCoV-2 Detect™ IgG (InBios) or SARS-CoV-2 Surrogate Virus Neutralization Test (GenScript), respectively, according to the manufacturer’s instructions. Peptide-specific ELISAs were performed using chemically synthesized N-terminally biotinylated 20-mer peptides (Sigma) with sequences from SARS-CoV-2 epitopes SH ([Btn]_LQPELDSFKEELDKYFKNHT) and SD2 ([Btn]_EVPVAIHADQLTPTWRVYSTGSNVFQTRAG). NeutrAvidin-coated plates (Pierce) were coated with peptide diluted to 5 μg/mL in PBS and incubated overnight at 4°C. Next, plates were blocked with 1X Blocker BSA (Thermo) in PBS for 1 h at room temperature (RT). Plasma samples diluted in Superblock TBST (Pierce) were added to wells and incubated for 1 h at 37°C. Goat anti-human IgG-HRP (Abcam) was applied to allwells and incubated for 30 min at 37°C. Ultra TMB (Thermo) was added and plates were incubated at RT in darkness for 12–15 min; the colorimetric reaction was stopped with sulfuric acid (Thermo). Plates were read on a BioRad xMark spectrophotometer at 450 nm. Plates were washed 6X with 1X TBST20 (Pierce) following each step save after TMB addition. Assay background was subtracted from sample wells prior to analysis.
6
Quantitative ELISA for Total IgG
7
Antigen-specific IgG Quantification by ELISA
8
Glycosylation Knockout in Arabidopsis
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Col-0 A. thaliana (plant-specific glycosylation knockout mutant TKO Arabidopsis), pPhasBar plasmid, and EHA105 Agrobacterium tumefaciens strains were supplied by Jilin Agriculture University Ministry of Education Engineering Research Center of Bioreactor and Pharmaceutical Development. Escherichia coli DH5α was obtained from Genewiz Biotechnology (Suzhou, China). Restriction endonucleases HindIII, NcoI, and XbaI were purchased from New England Biolabs (Ipswich, MA, USA). Polyvinylidene fluoride (PVDF) membranes were purchased from Millipore. Goat anti-human IgG-HRP was purchased from Abcam (USA). Oligonucleotide primers were acquired from Genewiz Biotechnology (Suzhou, China). All chemicals were of the highest purity available.
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