To quantify serum or supernatant human antibody, diluted samples were added to plates (Corning Incorporated) coated with goat anti-human IgG Fcγ fragment (Jackson) or goat anti-human IgM (Southern Biotech), washed, and blocked with 2% Bovine Serum Albumin (BSA, Fisher Scientific). Bound human Ig was detected using biotin-conjugated mouse anti-human IgG (BD Pharmingen) or biotin-conjugated mouse anti-human IgM (BD Pharmingen) secondary antibodies, followed by streptavidin-horseradish peroxidase (Thermo Scientific). Colorimetric change by 3,3′,5,5′-Tetramethylbenzidine substrate solution (Thermo Scientific) was stopped by 2M sulfuric acid (Sigma), and optical densities determined by spectrophotometer 450nm absorbance. Human serum with known IgM and IgG concentrations (Bethyl) established a standard.
Goat anti human igm
Manufactured by Southern Biotech
Sourced in United States
Goat anti-human IgM is a laboratory reagent used to detect and quantify human immunoglobulin M (IgM) in biological samples. It is a polyclonal antibody produced by immunizing goats with human IgM. The product is designed for research and diagnostic applications where the identification and measurement of human IgM is required.
Automatically generated - may contain errors
Lab products found in correlation
Bovine serum albumin (bsa), by Thermo Fisher Scientific (1 mentions)
Streptavidin horseradish peroxidase, by Thermo Fisher Scientific (1 mentions)
3 3 5 5 tetramethylbenzidine substrate solution, by Thermo Fisher Scientific (1 mentions)
2m sulfuric acid, by Merck Group (1 mentions)
Goat anti human igg fcγ fragment, by Jackson ImmunoResearch (1 mentions)
Nunc maxisorp elisa plate, by Thermo Fisher Scientific (1 mentions)
Goat anti human igm hrp, by Southern Biotech (1 mentions)
Anti jak2 d2e12, by Cell Signaling Technology (1 mentions)
Ibrutinib, by Selleck Chemicals (1 mentions)
Goat f ab 2 anti human igm, by Southern Biotech (1 mentions)
Human cxcl12, by R&D Systems (1 mentions)
Human icam 1 fc, by R&D Systems (1 mentions)
Human e selectin fc, by R&D Systems (1 mentions)
Fluorescein isothiocyanate fitc goat secondary antibody to mouse, by Merck Group (1 mentions)
Sirnas on targetplus smartpool, by Thermo Fisher Scientific (1 mentions)
Kim127 mouse monoclonal antibody, by American Type Culture Collection (1 mentions)
P473akt, by Cell Signaling Technology (1 mentions)
Ethylene glycol tetraacetic acid (egta), by Merck Group (1 mentions)
Phenyl methyl sulfonyl fluoride (pmsf), by Thermo Fisher Scientific (1 mentions)
Sodium chloride (nacl), by Merck Group (1 mentions)
6 protocols using goat anti human igm
1
Quantification of Human Serum Antibodies
2
Quantifying Polyreactive IgM in Transplanted Patients
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Nunc MaxiSorp ELISA plates (Thermo Fisher Scientific 44-2404-21) were coated overnight with goat anti–human IgM (4 μg/mL; SouthernBiotech 2020-01, RRID:AB_2795599). After blocking, the plates were incubated with serum from transplanted patients. Bound IgM was detected by adding HRP-conjugated goat anti–human IgM (4 μg/mL; SouthernBiotech 2020-05, RRID:AB_2795603). Alternatively, polyreactive natural IgM was detected by analyzing the amount of immunoglobulin bound to LPS adapted from the protocol Singh et al. described (59 (link)). Briefly, Nunc MaxiSorp ELISA plates were coated overnight at room temperature with LPS (10 μg/mL; Alpha Diagnostic International LPS12-1). After blocking, human sera were incubated for 2 hours at 37°C. Bound IgM was detected by adding goat anti–human IgM–HRP (4 μg/mL; SouthernBiotech 2020-05, RRID:AB_2795603). The reactions were visualized by subsequent addition of 2,2′-Azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) substrate (SouthernBiotech 0202-01). All readings were recorded at 405 nm.
3
Immune Cell Adhesion Protein Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human E-selectin/Fc, human ICAM-1/Fc, human VCAM-1/Fc and human CXCL12 were from R&D Systems (R&D Systems, Minneapolis, MN, USA); goat anti-human IgM was from SouthernBiotech (SouthernBiotech, Birmingham, AL, USA); fluorescein isothiocyanate (FITC) goat secondary antibody to mouse was from Sigma (Sigma-Aldrich, St. Louis, MO, USA); KIM127 mouse monoclonal antibody was from American Type Culture Collection (ATCC, Rockville, MD, USA); 327A mouse monoclonal antibody was kindly provided by Dr. Kristine Kikly (Eli Lilly and Co., Indianapolis, IN, USA); goat F(ab’)2 anti-human IgM was from Southern Biotech (Southern Biotech, Birmingham, AL, USA); Tyrphostin and rabbit polyclonal anti-actin antibody were from Sigma; Ibrutinib was from Selleck Chemicals (Selleck Chemicals, Houston, TX, USA); PE mouse anti-BTK (pY223) antibody was from BD Biosciences (BD Biosciences, San Jose, CA, USA); rabbit monoclonal anti-JAK2 (D2E12) was from Cell Signaling Technology (Danvers, MA, USA); siRNAs (ON-TARGETplus SMARTpool) were from Thermo Fisher Scientific (Fremont, CA, USA).
4
Ibrutinib Inhibition of BCR Signaling
5
Detection of IgM-Secreting B Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
On the fourth day of primary B cell culture, Immunolon® flat-bottom 96-well microtiter plates (ThermoFisher Scientific) were coated with goat antihuman IgM (Southern Biotech) at a final concentration of 10 µg/ml in HBSS for 1 h at 37°C. Plates were then washed and blocked with culture medium for 1 h at room temperature. Live cells were counted via trypan blue exclusion and seeded as serial 1:2 dilutions in fresh culture medium in the plate in duplicate. Plates were briefly spun to settle cells to the bottom and incubated overnight in a 37°C humidified 5% CO2 incubator. Plates were subsequently washed and biotinylated goat antihuman IgM was added at a final concentration of 1.6 µg/ml. The plates were incubated for 2 h at 37°C. Plates were washed again and alkaline phosphatase (AP)-conjugated streptavidin was added at a dilution of 1:1,000 and incubated for 1 h at 37°C. Plates were washed and developing solution was added: 1 mg/ml BCIP (Sigma) in AMP buffer. Color development took place at room temperature in the dark until spots appeared; plates were then rinsed with dH2O, dried and counted under a light microscope.
6
ZIKV Antibody Detection Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Falcon 96-well plates were precoated with 100 μl of ZIKV (1 × 1010 genome copies/ml) diluted in Tris-buffered saline (TBS;pH 7.4) overnight at 4°C. After blocking with TBS containing 5% skim milk was performed for 1 h at room temperature, 100 μl of plasma or purified IgG samples serially diluted in blocking buffer containing 0.05% Tween 20 was added and incubated for 1 h at room temperature, followed by 1:3,000-diluted horseradish peroxidase-conjugated goat anti-human IgG (Jackson Research Laboratories) or goat anti-human IgM (Southern Biotech) for 1 h. After washing with TBS containing 0.05% Tween 20, the reaction was developed by the use of the peroxidase substrate tetramethylbenzidine (Millipore) for 3 min and stopped by adding the same volume of 2 M phosphoric acid. The absorbance at 450 nm was read in SpectraMax Paradigm plate reader (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!