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Authors: Angel Justiz-Vaillant, Belkis Ferrer-Cosme
Group: University of the West Indies, [email protected]
Summary: Granulocyte macrophage-colony stimulating factor (GM-CSF) is a monomeric glycoprotein. It is a cytokine secreted by macrophages, T cells, natural killer cells, mast cells, endothelial cells and fibroblasts. It acts as a growth factor. [1] Reference1. Egea L, Hirata Y, Kagnoff MF. GM-CSF: a role in immune and inflammatory reactions in the intestine. Expert Rev Gastroenterol Hepatol. 2010 Dec;4(6):723-31. doi: 10.1586/egh.10.73. PMID: 21108592; PMCID: PMC3291482.
Proper citation: Angel Justiz-Vaillant, Belkis Ferrer-Cosme 2020. ELISA for quantification of granulocyte macrophage-colony stimulating factor (GM-CSF) in tissue culture supernatant, human serum or plasma.. protocols.io dx.doi.org/10.17504/protocols.io.bktykwpw Copy
Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Summary: Interleukins (IL) are a type of cytokine first thought to be expressed by leukocytes alone but have later been found to be produced by many other body cells. They play essential roles in the activation and differentiation of immune cells, as well as proliferation, maturation, migration, and adhesion. They also have pro-inflammatory and anti-inflammatory properties. The primary function of interleukins is, therefore, to modulate growth, differentiation, and activation during inflammatory and immune responses. Interleukins consist of a large group of proteins that can elicit many reactions in cells and tissues by binding to high-affinity receptors in cell surfaces. Phagocytes mainly make IL-36. It acts on T lymphocytes and NK cells regulating the IFN-γ synthesis. It stimulates the hematopoiesis and expression of both MHC class I and II molecules as well as intracellular adhesion molecules (ICAM)-1. [1]Reference1. Justiz Vaillant AA, Qurie A. Interleukin. In:StatPearls. Treasure Island (FL): StatPearls Publishing; June 12, 2019.
Proper citation: Angel Justiz-Vaillant 2020. ELISA for quantification of IL-36 in human serum.. protocols.io dx.doi.org/10.17504/protocols.io.bj38kqrw Copy
Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Summary: Keyhole limpet hemocyanin (KLH) is a cooper-containing protein comprising of subunits with MW of 400 kDa. This protein is found in the hemolymph of the sea mollusk Megathura crenulata. It has the ability to enhance the host’s immune response by interacting with monocytes, T cells and macrophages. KLH has been used primarily as a carrier for vaccines and antigens [1]. It was found that chicken immunized with KLH bound peptide raised an anti-KLH immunoresponse [2]. This can be tested by a single method such as the Ouchterlony technique.Reference1. Aarntzen EH, de Vries IJ, Göertz JH, et al. Humoral anti-KLH responses in cancer patients treated with dendritic cell-based immunotherapy are dictated by different vaccination parameters.Cancer Immunol Immunother. 2012;61(11):2003-2011. doi:10.1007/s00262-012-1263-z2. Justiz Vaillant AA, Anderson MF, Smikle M, Wisdom B, Mohammed W, et al. (2013) Development of Anti HIV Gp120 and HIV Gp41 Peptide Vaccines. J Vaccines Vaccin 4: 206. doi: 10.4172/2157-7560.1000206
Proper citation: Angel Justiz-Vaillant 2020. Detection of anti- SpA antibodies in egg white tested by double immunodiffusion (Ouchterlony) technique.. protocols.io dx.doi.org/10.17504/protocols.io.bjsqkndw Copy
Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Proper citation: Angel Justiz-Vaillant 2020. Immunoblot analyses for investigating SpLA binding to purified mammalian and avian immunoglobulins.. protocols.io dx.doi.org/10.17504/protocols.io.bjq7kmzn Copy
Authors: Angel Justiz-Vaillant, Monica F. Smikle
Group: University of the West Indies, [email protected]
Summary: Referencesst protein LA (SpLA) is an immunoglobulin-binding protein that interacts with the Fab and Fc regions of many mammalian immunoglobulins [1].References1.Vaillant AJ, McFarlane-Andersonv N, Wisdom B, Mohammed W, Vuma S, et al. (2013) Immunoglobulin-binding Bacterial Proteins (IBP) Conjugates and their Reactivity with Immunoglobulin in Enzyme-Linked Immunosorbent Assays (ELISA). J Anal Bioanal Tech 4: 175. doi:10.4172/2155-9872.1000175
Proper citation: Angel Justiz-Vaillant, Monica F. Smikle 2020. Direct ELISA for investigating the binding of recombinant Protein-LA to immunoglobulins.. protocols.io dx.doi.org/10.17504/protocols.io.bjxukpnw Copy
Authors: Angel Justiz-Vaillant, Monica F. Smikle
Group: University of the West Indies, [email protected]
Proper citation: Angel Justiz-Vaillant, Monica F. Smikle 2020. Direct ELISA for investigating the binding of Protein-A to immunoglobulins. . protocols.io dx.doi.org/10.17504/protocols.io.bjxqkpmw Copy
Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Summary: Interleukins (IL) are a type of cytokine first thought to be expressed by leukocytes alone but have later been found to be produced by many other body cells. They play essential roles in the activation and differentiation of immune cells, as well as proliferation, maturation, migration, and adhesion. They also have pro-inflammatory and anti-inflammatory properties. The primary function of interleukins is, therefore, to modulate growth, differentiation, and activation during inflammatory and immune responses. Interleukins consist of a large group of proteins that can elicit many reactions in cells and tissues by binding to high-affinity receptors in cell surfaces. It is strongly associated inflammatory activity with IL-26. Th17 cells produce this interleukin. It acts on epithelial cells and intestinal epithelial cells. It induces IL-10 expression, stimulates the production of IL-1-beta, IL-6, and IL-8 and causes Th17 cell generation. [1]Reference1. Justiz Vaillant AA, Qurie A. Interleukin. In:StatPearls. Treasure Island (FL): StatPearls Publishing; June 12, 2019.
Proper citation: Angel Justiz-Vaillant 2020. ELISA for quantification of IL-26 in human serum.. protocols.io dx.doi.org/10.17504/protocols.io.bj3vkqn6 Copy
Authors: Angel Justiz-Vaillant
Group: Carbon
Summary: This ELISA was used to study the interactions between SpG) and SpLG) with different immunoglobulin preparations from mammalian and avian species.
Proper citation: Angel Justiz-Vaillant 2021. Streptococcal Protein-G and Protein-LG sandwich ELISA. protocols.io dx.doi.org/10.17504/protocols.io.btaznif6 Copy
Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Summary: Interleukins (IL) are a type of cytokine first thought to be expressed by leukocytes alone but have later been found to be produced by many other body cells. They play essential roles in the activation and differentiation of immune cells, as well as proliferation, maturation, migration, and adhesion. They also have pro-inflammatory and anti-inflammatory properties. The primary function of interleukins is, therefore, to modulate growth, differentiation, and activation during inflammatory and immune responses. Interleukins consist of a large group of proteins that can elicit many reactions in cells and tissues by binding to high-affinity receptors in cell surfaces. Different cells in both innate and acquired immunities produce IL-22, but the primary sources are T cells. Th22 cell is a new line of CD4+ T cells, which differentiated from naive T cells in the presence of various pro-inflammatory cytokines including IL-6. IL-22 inhibits IL-4 production. It also has essential functions in mucosal surface protection and tissue repairy. [1]Reference1. Justiz Vaillant AA, Qurie A. Interleukin. In:StatPearls. Treasure Island (FL): StatPearls Publishing; June 12, 2019.
Proper citation: Angel Justiz-Vaillant 2020. ELISA for quantification of IL-22 in human serum.. protocols.io dx.doi.org/10.17504/protocols.io.bj3rkqm6 Copy
Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Summary: Peroxidase labeled anti-human IgG is a conjugated secondary antibody that can be used to detect specific antibodies against millions of antigens in humans. This reagent is usually used in enzyme-linked immunosorbent assay, immunoblot analysis and dot blot [1,2]. Reference1. Stubenrauch K, Wessels U, Essig U, Kowalewsky F, Vogel R, Heinrich J. Characterization of murine anti-human Fab antibodies for use in an immunoassay for generic quantification of human Fab fragments in non-human serum samples including cynomolgus monkey samples.J Pharm Biomed Anal. 2013;72:208-215. doi:10.1016/j.jpba.2012.08.0232.Justiz Vaillant AA, McFarlane-Anderson N, Akpaka PE, Smikle MP, Ramirez N, et al. (2013) Use of Dot Blots Analysis in the Separation of Anti-HIV Antibodies in Animals. J Chromat Separation Techniq 4: 181. doi:10.4172/2157- 7064.1000181
Proper citation: Angel Justiz-Vaillant 2020. Protocol of preparation of horseradish peroxidase (HRP) conjugated to anti-human IgG to be used as secondary antibody in immunoassays. . protocols.io dx.doi.org/10.17504/protocols.io.bjmxkk7n Copy
Authors: Angel Justiz-Vaillant
Group: Carbon
Summary: This ELISA was used to study the interactions between protein-LAG (PLAG) and streptococcal protein-G (SpG) with different immunoglobulin preparations of mammalian and avian species.
Proper citation: Angel Justiz-Vaillant 2021. Chimeric Protein-LAG and Streptococcal protein G sandwich ELISA. protocols.io dx.doi.org/10.17504/protocols.io.btbcniiw Copy
Authors: Angel Justiz-Vaillant, Monica F. Smikle
Group: University of the West Indies, [email protected]
Proper citation: Angel Justiz-Vaillant, Monica F. Smikle 2020. Universal sandwich ELISA for investigating the binding of avian immunoglobulins to Streptococcal protein-G (SpG) using anti-IgY-peroxidase conjugate. . protocols.io dx.doi.org/10.17504/protocols.io.bjqvkmw6 Copy
Authors: Angel Justiz-Vaillant, Belkis Ferrer-Cosme
Group: University of the West Indies, [email protected]
Proper citation: Angel Justiz-Vaillant, Belkis Ferrer-Cosme 2020. ELISA for measurement of serum macrophage migration inhibitory factor (MIF). . protocols.io dx.doi.org/10.17504/protocols.io.bj7ekrje Copy
Authors: Angel Justiz-Vaillant
Group: Carbon
Summary: This ELISA was used to study the interactions between protein-LAG (PLAG) and staphylococcal protein-A (SpA) with different immunoglobulin preparations of mammalian and avian species.
Proper citation: Angel Justiz-Vaillant 2021. Chimeric Protein-LAG and Staphylococcal Protein A sandwich ELISA. protocols.io dx.doi.org/10.17504/protocols.io.btbbniin Copy
Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Proper citation: Angel Justiz-Vaillant 2020. Purification of avian egg yolk immunoglobulins using the water dilution method and precipitation with isopropanol. . protocols.io dx.doi.org/10.17504/protocols.io.bjvpkn5n Copy
Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Summary: Interleukin-10 is a pleiotropic cytokine playing a critical role as a regulator of myeloid and lymphoid cell function. Due to the ability of IL-10 to blocking cytokine synthesis and many accessory cell functions of antigen-presenting cells such as macrophages this cytokine is a potent suppressor of the effector functions of macrophages, T-cells and NK cells. IL-10 also participates in regulating proliferation and differentiation of B-lymphocytes, mast cells and thymocytes [1]. Reference1. Ouyang W, O'Garra A. IL-10 Family Cytokines IL-10 and IL-22: from Basic Science to Clinical Translation.Immunity. 2019;50(4):871-891. doi:10.1016/j.immuni.2019.03.020
Proper citation: Angel Justiz-Vaillant 2020. ELISA for quantification of IL-6 in human serum or plasma.. protocols.io dx.doi.org/10.17504/protocols.io.bjy7kpzn Copy
Authors: Angel Justiz-Vaillant
Summary: This reagent can be used in ELISA, Western blotting and Dot blot to detect antigens and antibodies. It is important in the immunodiagnosis of infectious diseases and other problems. I find this useful in the detection of anti-HIV antibodies by ELISA.
Proper citation: Angel Justiz-Vaillant 2020. Preparation of staphylococcal protein-A conjugated to horseradish peroxidate by the periodate method.. protocols.io dx.doi.org/10.17504/protocols.io.bjg8kjzw Copy
Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Proper citation: Angel Justiz-Vaillant 2020. An easy chromatographic method for purification of Immunoglobulin Y (IgY) using HiTrap™ Columns.. protocols.io dx.doi.org/10.17504/protocols.io.bju7knzn Copy
Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Summary: Chemical synthesis facilitates the generation of peptides which are difficult to express in bacteria. The fragment 254-274 of the human immunodeficiency virus (HIV) is critical for infectivity of the virus and induction of antibody neutralization (Ho et al, 1988). Boudet et al, 1995 demonstrated that a synthetic peptide derived from the third variable domain of the HIV-1 gp-120 when used as immunogen was able to induce an antibody response to multiple (up to six) HIV strains. ReferenceHo DD, Kaplan JC, Rackauskas IE, Gurney ME. Second conserved domain of gp120 is important for HIV infectivity and antibody neutralization.Science. 1988;239(4843):1021-1023. doi:10.1126/science.2830667.Boudet F, Keller H, Kieny MP, Thèze J. Single peptide and anti-idiotype based immunizations can broaden the antibody response against the variable V3 domain of HIV-1 in mice.Mol Immunol. 1995;32(7):449-457. doi:10.1016/0161-5890(95)00007-2
Proper citation: Angel Justiz-Vaillant 2020. Conjugation of Keyhole limpet haemocynin to Peptide 254-274 of HIV gp-120 as immunogen.. protocols.io dx.doi.org/10.17504/protocols.io.bjh7kj9n Copy
Authors: Angel Justiz-Vaillant, Belkis Ferrer-Cosme
Group: University of the West Indies, [email protected]
Summary: IgD is a monomer with a molecular weight of 184 Kd. IgD is present in a meager amount in the serum (0.03 mg/mL) and has an unknown function against pathogens. It is regarded as a BCR. IgD may play an essential role in antigen-triggered lymphocyte differentiation. [1]
Proper citation: Angel Justiz-Vaillant, Belkis Ferrer-Cosme 2020. ELISA for quantification of human immunoglobulin D (IgD) in serum or plasma.. protocols.io dx.doi.org/10.17504/protocols.io.bj7wkrpe Copy
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