Ab195215

Manufactured by Abcam

Sourced in United States, United Kingdom

Ab195215 is a mouse monoclonal antibody that recognizes a specific protein target. It is designed for use in laboratory research applications.

Automatically generated - may contain errors

Lab products found in correlation

3α aminocholestane 3ac, by Echelon Biosciences (1 mentions) El312 bio kinetics microplate reader, by Agilent Technologies (1 mentions) Ab269370, by Abcam (1 mentions) Ab108792, by Abcam (1 mentions) Ab196263, by Abcam (1 mentions) 96 well half area plate, by Corning (1 mentions) Edta collection tube, by BD (1 mentions) Superblock buffer, by Thermo Fisher Scientific (1 mentions) Recombinant ebov gp, by IBT Bioservices (1 mentions) Ab157719, by Abcam (1 mentions)

4 protocols using ab195215

IVIG and SHIP1 Inhibitor Therapy for Intracerebral Hemorrhage

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IVIG (Gammagard Liquid-Baxter) was administrated intraperitoneally (i.p), one hour after ICH. Two doses (0.5 and 2.0 g/kg) were tested. The effectivity of intraperitoneal injection for high molecular weight drug (molecular weight of IVIG is ~ 300 kDa) was tested by ELISA (Abcam, ab 195215). Animals were treated with 0.5 or 2,0 g/kg. 24 hours after drug administration animals were anesthetized and blood were collected using inferior vena cava. The level of human IgG (active component of IVIG) in mice was investigated according to vendor’s recommendation. 6 animals per group and 6 naïve animals were used
SHIP1 inhibitor, 3α-aminocholestane (3AC), (Echelon Biosciences, 30 mg/kg) was administrated 30 minutes post-ICH, intraperitoneally. siRNA FcγRIIB or control (scrambled) RNA (OriGene Technologies, 100 pmol/2 μL) was given via intracerebroventricular (i.c.v) injection 24 hours before ICH.

Akyol G.Y., Manaenko A., Akyol O., Solaroglu I., Ho W.M., Ding Y., Flores J., Zhang J.H, & Tang J. (2017). IVIG activates FcγRIIB-SHIP1-PIP3 Pathway to stabilize mast cells and suppress inflammation after ICH in mice. Scientific Reports, 7, 15583.

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Quantifying Secreted SA-IgG by ELISA

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The level of SA-IgG in cell supernatant was determined using enzyme-linked immunosorbent assay (ELISA) according to the slightly modified procedure (ab195215; Abcam, USA). Assay diluent cell culture supernatants were added into appropriate wells and incubated for 2.5 h at room temperature. After washing with wash solution, the plate was added 100 μL of 1:50 diluted SNA (VECTOR, USA) for 2 h. HRP-Streptavidin solution was incubated for 45 mins. After TMB (3,3′, 5,5;-tetramethylbenzidine) solution incubation for 30 mins, the reaction was stopped with a stop solution. The optical densities (ODs) were determined at 450 nm with an EL312 Bio-Kinetics microplate reader (Bio-TekInstruments, Winooski, VT).

Liu Y., Yu H., Wu S., Yang X., Cao C., Wang F., Jia J, & Yan T. (2021). Plasma ST6GAL1 regulates IgG sialylation to control IgA nephropathy progression. Therapeutic Advances in Chronic Disease, 12, 20406223211048644.

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Cytokine Profiling in Mouse Xenograft Serum

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The mouse serums obtained from xenogeneic transplantation were assayed for 27 cytokines using the Bio-Plex Suspension Array System with Bio-Plex Pro Human Cytokine Screening 27-Plex Panel (Bio-Rad Laboratories Inc., CA, USA). The assay was performed for the following cytokines: IL-1β, IL-1Ra, IL-2, IL-4, IL-5, IL-6, IL-7, IL-9, IL-10, IL-12p70, IL-13, IL-15, IL-17, Eotaxin, FGF basic, GM-CSF, IFN-γ, IP-10, MIP-1a, PDGF-BB, MIP-1b, RANTES, TNF-alpha, and CXCL13, which were measured using a commercially available specific ELISA kit (ab269370, Abcam) according to the manufacturer’s instructions. The values of each cytokine were used to generate heat maps. Human IgG, IgA, IgE and mouse albumin levels in mouse serum were measured using a commercially available ELISA kit (ab195215, ab196263, ab195216, and ab108792, Abcam) according to the manufacturer’s instructions.

Harada T., Kikushige Y., Miyamoto T., Uno K., Niiro H., Kawakami A., Koga T., Akashi K, & Yoshizaki K. (2023). Peripheral helper-T-cell-derived CXCL13 is a crucial pathogenic factor in idiopathic multicentric Castleman disease. Nature Communications, 14, 6959.

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EBOV Antibody Quantification Protocol

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Serum samples were collected by saphenous bleed in EDTA collection tubes (BD, Franklin Lakes, New Jersey) and aliquoted for storage at −80°C. Human IgG and murine IgG concentrations were quantified using commercially available kits (Abcam, Cambridge, United Kingdom, ab195215 and ab157719). Reciprocal antibody titers were determined by coating half-area 96-well plates (Corning, New York) with 1 μg/mL recombinant EBOV GP (IBT Bioservices, Rockville, DC, 0501-001), EBOV VP40 (IBT Bioservices, Rockville, DC, 0564-001), or Influenza A virus HA (SinoBiological Beijing, China, 11,684-V08H) protein overnight at 4°C. Plates were washed four times with 0.2% PBS-Tween20 (PBS-T) and blocked with SuperBlock buffer (Fisher, Waltham, Massachusetts). 2-fold serial dilutions were incubated at 37°C for 1 h and then washed four times with PBS-T. Secondary antibody (Pierce P31430) was incubated at 1:2,000 for 1 h at 37°C, washed four times with PBS-T, and incubated with TMB substrate (Pierce PI34021) for 15 min before reading absorbance values at 650 nm. Reciprocal titer was defined as the highest 2-fold serum dilution that gave an OD₆₅₀ value 2-fold greater than negative control wells.

van Lieshout L.P., Rghei A.D., Cao W., He S., Soule G., Zhu W., Thomas S.P., Sorensen D., Frost K., Tierney K., Thompson B., Booth S., Safronetz D., Kulkarni R.R., Bridle B.W., Qiu X., Banadyga L, & Wootton S.K. (2022). AAV-monoclonal antibody expression protects mice from Ebola virus without impeding the endogenous antibody response to heterologous challenge. Molecular Therapy. Methods & Clinical Development, 26, 505-518.

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