Pooled complement human serum

Manufactured by Innovative Research

Pooled complement human serum is a laboratory product derived from the blood of multiple human donors. It contains a complex mixture of proteins, including the complement system components, which play a crucial role in the immune response. This product is commonly used in various immunological and biochemical research applications.

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Lab products found in correlation

Medium 199, by Lonza (2 mentions) Nitrocellulose membranes for immunoblotting, by Bio-Rad (2 mentions) β actin, by Cell Signaling Technology (2 mentions) Laemmli sample buffer, by Bio-Rad (2 mentions) Control igg, by Santa Cruz Biotechnology (1 mentions) Total p 70 s6, by Cell Signaling Technology (1 mentions) Dulbecco modified eagle medium (dmem), by GE Healthcare (1 mentions) Egm 2, by Lonza (1 mentions) Fetal bovine serum (fbs), by Welgene (1 mentions) Human umbilical vein endothelial cells huvecs, by Lonza (1 mentions) Chlorpromazine, by Merck Group (1 mentions) Heparin, by Merck Group (1 mentions) Phosphonoformic acid, by Merck Group (1 mentions) Ethylene glycol tetraacetic acid (egta), by Merck Group (1 mentions) Gw4869, by Merck Group (1 mentions) Methyl β cyclodextrin, by Merck Group (1 mentions) Ethylene diamine tetraacetic acid (edta), by Merck Group (1 mentions) Egm 2 bulletkit, by Lonza (1 mentions) Huvecs, by Lonza (1 mentions) Cba human anaphylatoxin kit, by BD (1 mentions)

5 protocols using pooled complement human serum

Antibodies for Runx2, MAPK, and ATPase

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Antibodies against Runx2, Phospho-p44/42 MAPK (p-Erk1/2), total p44/42 MAPK (Erk1/2), Phospho P-70 S6, total P-70 S6, and β-actin were purchased from Cell Signaling, Inc (Beverly, Massachusetts). Antibody against H-K-ATPase is from Lifespan Biosciences Inc. H-K-ATPase neutralization antibody is from Abcam, and control IgG is from Santa Cruz. Pooled complement human serum was from Innovative Research (Novi, Michigan). Medium 199 was purchased from Lonza (Walkersville, Maryland). Laemmli sample buffer and nitrocellulose membranes for immunoblotting were purchased from BioRad (Hercules, California). MAPK inhibitor UO126 is from MCE. Trypsin-EDTA solution (0.25%), H-K-ATPase inhibitor Ouabain and Sch28080, and all other chemicals were purchased from Sigma–Aldrich Chemical Co (St. Louis, Missouri).

Münk C., Löhler J., Prassolov V., Just U., Stockschläder M, & Stocking C. (1997). Amphotropic murine leukemia viruses induce spongiform encephalomyelopathy. Proceedings of the National Academy of Sciences of the United States of America, 94(11), 5837-5842.

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Cell Culture Protocols for Cellular Assays

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U2-OS, HeLa, and T24 cells were obtained from the Korean Cell Line Bank (Seoul, South Korea). The cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM; GE Healthcare, Little Chalfont, UK) supplemented with 10% fetal bovine serum (FBS; Welgene, Seoul, South Korea) and 1% antibiotics (Lonza, Allendale, NJ). Human umbilical vein endothelial cells (HUVECs) were purchased from Lonza and cultured in endothelial cell growth medium-2 (EGM-2; Lonza) bullet kit. The cells were maintained in a humidified atmosphere of 5% CO₂ at 37 °C. Pooled complement human serum was purchased from Innovative Research, Inc (Novi, MI) and used as normal human serum (NHS) in all experiments. Heat-inactivation was performed using this serum at 56 °C for 30 min and used as heat-inactivated human serum (HHS).

Jeon H., Han S.R., Lee S., Park S.J., Kim J.H., Yoo S.M, & Lee M.S. (2018). Activation of the complement system in an osteosarcoma cell line promotes angiogenesis through enhanced production of growth factors. Scientific Reports, 8, 5415.

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Immunoblotting Assay for Bone and Inflammatory Markers

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Antibodies against collagen I, collagen III and MMP-9 were purchased from Boster Biological technology Co (Pleasanton, CA). Antibodies against Runx-2, Phospho-p44/42 MAPK (p-Erk1/2), total p44/42 MAPK (Erk1/2), ICAM 1, and β-actin were purchased from Cell Signaling, Inc (Beverly, MA). Antibody against complement C3 was from St John’s Laboratory (London, UK). Pooled complement human serum was from Innovative Research (Novi, MI). Medium 199 was purchased from Lonza (Walkersville, MD). Recombinant human C3a and C5a are purchased from R&D Systems (Minneapolis, MN). Laemmli sample buffer and nitrocellulose membranes for immunoblotting were purchased from Bio-Rad (Hercules, CA). Trysin-EDTA solution (0.25%) and all other chemicals were purchased from Sigma-Aldrich Chemical Co (St. Louis, MO).

Deng X.S., Meng X., Fullerton D., Stone M, & Jaggers J. (2021). Complement up-regulates Runx-2 to induce pro-fibrogenic change in aortic valve interstitial cells. The Annals of thoracic surgery, 112(6), 1962-1972.

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Studying Complement-Mediated Endothelial Responses

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HUVECs were purchased from Lonza (Allendale, NJ), and cultured with endothelial cell growth medium-2 (EGM-2) bullet kit (Lonza) in a humidified atmosphere of 5% CO₂ at 37°C. Pooled complement human serum was purchased from Innovative Research, Inc (Novi, MI) and used as normal human serum (NHS) in all experiments. Heat-inactivation was performed with this serum at 56°C for 30 min. Factor B-depleted human serum, C3-depleted human serum, C6-depleted human serum, purified properdin, purified C3 and purified factor B were purchased from Quidel Corporation (San Diego, CA). Factor P (properdin)-depleted human serum was purchased from Complement Technology, Inc. (Tyler, Tx). EDTA, EGTA, heparin, GW4869, methyl-β–cyclodextrin, chlorpromazine, and phosphonoformic acid were obtained from Sigma-Aldrich (St. Louis, MO).

Jeon H., Yoo S.M., Choi H.S., Mun J.Y., Kang H.G., Lee J., Park J., Gao S.J, & Lee M.S. (2017). Extracellular vesicles from KSHV-infected endothelial cells activate the complement system. Oncotarget, 8(59), 99841-99860.

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Nanocarrier Complement Activation Assay

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Nanocarrier formulations (10 mg mL⁻¹) were added to pooled complement human serum (Innovative Research) in a 1:20 ratio and incubated at 37 °C, 200 rpm for 1 h. Afterward, samples were treated with 5 × 10⁻³ m EDTA to halt complement activation and were placed on ice. The C3a, C4a, and C5a human anaphylatoxin concentrations were quantitatively assessed using a BD™ CBA Human Anaphylatoxin Kit. The C3a, C4a, and C5a standards and were prepared following the manufacturer’s protocol. Data acquisition was completed using a BD LSRFortessa. Analysis proceeded using Cytobank software.

Vincent M.P., Karabin N.B., Allen S.D., Bobbala S., Frey M.A., Yi S., Yang Y, & Scott E.A. (2021). The Combination of Morphology and Surface Chemistry Defines the Immunological Identity of Nanocarriers in Human Blood. Advanced therapeutics, 4(4), 2100062.

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