Normal goat serum ngs

Manufactured by Merck Group

Sourced in United States

Normal goat serum (NGS) is a laboratory reagent derived from the blood of healthy goats. It is a complex mixture of proteins, antibodies, and other biomolecules that can be used as a blocking or dilution agent in various immunoassay and cell culture applications.

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Lab products found in correlation

Bovine serum albumin (bsa), by Merck Group (3 mentions) Axio observer z1, by Zeiss (3 mentions) Lsm 710 confocal microscope, by Zeiss (3 mentions) Microplate reader, by Bio-Rad (2 mentions) Polysorp 96 well plate, by Thermo Fisher Scientific (2 mentions) Vectashield mounting medium, by Vector Laboratories (2 mentions) Vectashield, by Vector Laboratories (2 mentions) Glial fibrillary acidic protein (gfap), by Merck Group (2 mentions) Normal goat serum (ngs), by Thermo Fisher Scientific (2 mentions) Neuronal nuclei (neun), by Merck Group (2 mentions) Digitonin, by Merck Group (1 mentions) Sodium pyruvate, by Thermo Fisher Scientific (1 mentions) N hydroxysuccinimide nhs, by Thermo Fisher Scientific (1 mentions) 1 ethyl 3 3 dimethylaminopropyl carbodiimide edc, by Thermo Fisher Scientific (1 mentions) Penicillin streptomycin, by Corning (1 mentions) L glutamine, by Thermo Fisher Scientific (1 mentions) Dulbecco s modified eagle medium dmem, by Merck Group (1 mentions) Fluoromount mounting medium, by Merck Group (1 mentions) Lsm510 meta system confocal microscope, by Zeiss (1 mentions) Aim 4, by Zeiss (1 mentions)

28 protocols using normal goat serum ngs

HeLa Cell Culture Conditions

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HeLa cells were cultured at 37°C and 5% CO₂ in Dulbecco’s modified Eagle medium (DMEM, Sigma Aldrich) supplemented with 5% fetal bovine serum (FBS, Sigma Aldrich), 1% penicillin-streptomycin (Cellgro), 2 mM L-glutamine (Thermo Fisher Scientific), and 1 mM sodium pyruvate (Thermo Fisher Scientific).
Bovine serum albumin (BSA), 2-(N-morpholino) ethane sulfonic acid (MES) buffer, normal goat serum (NGS), and digitonin were purchased from Sigma-Aldrich Corporation. N-hydroxysuccinimide (NHS), 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC), and all other chemicals were purchased from Thermo Fisher Scientific.

Sadr Karimi S, & Pante N. (2019). Carbon nanotubes as molecular transporters to study a new mechanism for molecular entry into the cell nucleus using actin polymerization force. PLoS ONE, 14(8), e0221562.

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Primate Brain Immunohistochemistry of TAT2A Peptide

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5(6)-FAM-conjugated TAT2A peptide was administrated to adult male non-human primates (i.p.) 30 min before sacrifice. Control monkeys were injected with the vehicle. The brain was dissected and post-fixed in 4% paraformaldehyde (PFA) in 0.1 M tris buffered saline (TBS) followed by sequential sinking in 20% and 30% sucrose in TBS. The tissue was then sectioned using a microtome and 40 μm-thick slices were stored in a cryoprotective solution at −20°C until use. Antigens were retrieved with 10 mM sodium citrate pH 6. Tissue sections were permeabilized in TBS containing 0.4% Triton X-100 (0.4% T-TBS) for 30 min at 4°C. After blocking in 5% Normal Goat Serum (NGS, #G9023, Sigma-Aldrich) in 0.1% T-TBS for 2 h at RT, slices were incubated with anti-HIV1 TAT and anti-microtubule associated protein 2 (MAP2) antibodies in 3% NGS-0.1% T-TBS overnight at 4°C. Tissue sections were then incubated with the appropriate AlexaFluor secondary antibodies in 3% NGS-TBS for 2 h at RT. Nuclei were stained with the fluorescent dye 4′,6-diamidino-2-phenylindole (DAPI, Life Technologies, Carlsbad, CA, USA). Slices were finally mounted with Fluoromount mounting medium (Sigma-Aldrich) onto glass slides. Labeling was visualized with LSM510 Meta system confocal microscope (Zeiss, Oberkochen, Germany) and Aim 4.2 software (Zeiss).

Mellone M., Stanic J., Hernandez L.F., Iglesias E., Zianni E., Longhi A., Prigent A., Picconi B., Calabresi P., Hirsch E.C., Obeso J.A., Di Luca M, & Gardoni F. (2015). NMDA receptor GluN2A/GluN2B subunit ratio as synaptic trait of levodopa-induced dyskinesias: from experimental models to patients. Frontiers in Cellular Neuroscience, 9, 245.

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Flow Cytometric Analysis of Cell Surface and Intracellular Markers

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Cells were kept at 4°C in 2% (v/v) paraformaldehyde (PFA, from Sigma). For surface staining, approximately 5x10⁵ cells were resuspended in 100 μL of FACS buffer (3% (v/v) Fetal Bovine Serum (FBS, from Invitrogen) in PBS) with the diluted primary antibody, and incubated for 15 min at room temperature in the dark. Cells were washed twice with PBS and resuspended in 300 μL of PBS to be analysed by flow cytometry (FACSCalibur, Becton Dickinson). For negative controls, cells were incubated with the appropriate isotypes. For intracellular staining, the protocol used is described by Miranda et al. [29 (link)]. For the negative controls, cells were incubated only with 3% (v/v) Normal Goat Serum (NGS, from Sigma) in PBS. The CellQuest software (Becton Dickinson) was used for all acquisition/analyses.

Badenes S.M., Fernandes T.G., Cordeiro C.S., Boucher S., Kuninger D., Vemuri M.C., Diogo M.M, & Cabral J.M. (2016). Defined Essential 8™ Medium and Vitronectin Efficiently Support Scalable Xeno-Free Expansion of Human Induced Pluripotent Stem Cells in Stirred Microcarrier Culture Systems. PLoS ONE, 11(3), e0151264.

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Serum IgM Antibody Detection by ELISA

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Serum IgM antibodies to PGL-I were detected by ELISA performed as described previously [31 (link)]. PolySorp 96-well plates (Nunc, Roskilde, Denmark) were coated with 0.01 μg/mL of the semi-synthetic analogue of PGL-I (NT-P-BSA; batch: Nara XVI-61, Dr Fujiwara, Japan), or BSA and blocked with PBS-Tween containing 1% BSA. Serum samples diluted 1/300 in PBS-T containing 10% normal goat serum/NGS (Sigma-Aldrich, St. Louis, USA) were incubate and washing, horse radish peroxidase/HRP-conjugated anti-human IgM (Immuno Chemicals, St. Louis, Missouri, USA) was added. After incubation and washing, peroxidase color substrate (TMB, Sigma Aldrich, St. Louis, USA), was added and the reaction was quenched by the addition of 2.5 N H₂SO₄, when the optical density/OD at 450 nm from reference serum reached an OD value of 0.6 (Bio-Rad microplate reader, Life Science, Hercules, CA, USA). The final OD was calculated by subtracting the OD values of BSA coated wells from OD values of NT-P-BSA wells. The cut-off was defined as OD > 0.25 as previously described [16 (link)].

Hungria E.M., Bührer-Sékula S., de Oliveira R.M., Aderaldo L.C., Pontes A.D., Cruz R., Gonçalves H.D., Penna M.L., Penna G.O, & Stefani M.M. (2017). Leprosy reactions: The predictive value of Mycobacterium leprae-specific serology evaluated in a Brazilian cohort of leprosy patients (U-MDT/CT-BR). PLoS Neglected Tropical Diseases, 11(2), e0005396.

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Microscopy of Recombinant GluN1-1b in HEK293T

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HEK293T cells (50,000) cultured at 37 °C/8% CO₂ in DMEM (high glucose, Life Technologies, Carlsbad, USA) were seeded on a 35 -mm dish, grown for 3 days, and transfected with 3 µg of myc-His-tagged GluN1-1b cloned into pcDNA4/TO/myc-His A (Invitrogen, Carlsbad, USA) using Metafectene-Pro (Biontex, Munich, Germany) [10 (link)]. One day post transfection, cells were split onto five poly-D-lysine-coated coverslips in a 35 -mm dish and 1 day later, they were fixed with 5% paraformaldehyde (PFA) for 20 min, washed 5× (PBS), permeabilized with 0.1% Triton X-100 for 5 min, again washed 5× (PBS), and blocked with 5% normal goat serum (NGS; Sigma-Aldrich, Munich, Germany) for 1 h. After five PBS washes, cells were incubated with serum and monoclonal mouse anti-myc IgG (clone 9E10, Hollmann-Lab, Bochum) for 1 h, washed with 10× (PBS), incubated for 1 h with fluorescein-labeled goat anti-monkey IgM (072-11-031; KPL, Gaithersburg, USA) and AlexaFluor®594-labeled goat anti-mouse IgG (A11005; Thermo Fisher) secondary AB, and PBS washed 5×. Cells were mounted in Fluoromount-G (Southern Biotech, Birmingham, USA) and analyzed via TCS-SP2-AOBS confocal microscope (63× oil immersion objective; Leica-Microsystems, Wetzlar, Germany). The results were independently assessed by three investigators.

Pan H., Oliveira B., Saher G., Dere E., Tapken D., Mitjans M., Seidel J., Wesolowski J., Wakhloo D., Klein-Schmidt C., Ronnenberg A., Schwabe K., Trippe R., Mätz-Rensing K., Berghoff S., Al-Krinawe Y., Martens H., Begemann M., Stöcker W., Kaup F.J., Mischke R., Boretius S., Nave K.A., Krauss J.K., Hollmann M., Lühder F, & Ehrenreich H. (2018). Uncoupling the widespread occurrence of anti-NMDAR1 autoantibodies from neuropsychiatric disease in a novel autoimmune model. Molecular Psychiatry, 24(10), 1489-1501.

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Cell Culture Media and Reagents

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Minimum Essential Medium alpha (MEMα), penicillin-streptomycin (P/S), bovine serum albumin (BSA), dimethyl sulfoxide (DMSO), Insulin-Transferrin-Selenium (ITS), and Tween-20, were purchased from Fischer Scientific (Waltham, MA). Perfluorooctanoic acid (PFOA; purity 95%) and normal goat serum (NGS) were purchased from Sigma-Aldrich (St. Louis, MO).

Clark K.L, & Davis J.S. (2022). Perfluorooctanoic acid (PFOA) promotes follicular growth and alters expression of genes that regulate the cell cycle and the Hippo pathway in cultured neonatal mouse ovaries. Toxicology and applied pharmacology, 454, 116253.

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Lectin and Immunohistochemistry Double Staining

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For
lectin and immunohistochemistry double staining, immediately after
the final TBS wash step in the lectin histochemistry method (as above)
and continuing the staining procedures in the dark at room temperature,
the spinal cord sections were rehydrated in 0.01 M PBS and then blocked
with 20% normal goat serum (NGS) (Sigma-Aldrich Co., Dublin, Ireland)
in PBS containing 0.2% Triton X-100 for 20 min. The primary antibodies
rabbit polyclonal anti-GFAP, 1:300 dilution (DakoCytomation, Dublin,
Ireland), rabbit polyclonal antineurogranin (1:100), mouse monoclonal
anti-βI-integrin (1:100), and rabbit polyclonal antigrowth-associated
protein-43 (GAP-43, 1:100) were diluted in PBS containing 2% NGS and
0.02% Triton X-100 and sections were incubated with the primary antibody
for 2 h. The sections were washed three times in PBS and the appropriate
secondary antibody, antirabbit IgG conjugated to rhodamine, or antimouse
conjugated to Alexa Fluor 594 (Life Technologies, Grand Island, NY)
was diluted 1:500 in PBS and incubated for 1 h. The sections were
again washed in PBS and cover slipped with ProLong Gold antifade reagent.
A negative control was carried out for each antibody by substituting
PBS for the primary antibody. Images were captured on an Olympus IX81
fluorescent microscope at 20× and 40× magnifications and
stored digitally for further image analysis.

Kilcoyne M., Patil V., O’Grady C., Bradley C, & McMahon S.S. (2019). Differential Glycosylation Expression in Injured Rat Spinal Cord Treated with Immunosuppressive Drug Cyclosporin-A. ACS Omega, 4(2), 3083-3097.

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Immunohistochemical Analysis of Testis in Cdk2-Deficient Mice

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Testes were collected from wild-type and Cdk2^-/- mice at P15, fixed in 4% paraformaldehyde, and embedded in paraffin. Section (5 μm-thick) were deparaffinized through xylene and graded ethanol dilutions, followed by antigen retrieval by microwaving in PBS buffer (5 mM Tris pH 8.0, 1 mM EDTA). After blocking with 5% normal goat serum (NGS; Sigma) and 0.2% Triton X-100 in PBS for 1 hr at RT, sections were incubated with primary antibodies for 2 hrs in PBS containing 5% NGS and 0.2% Triton-X100 at RT, washed with PBS, then incubated with secondary antibodies (Alexa 568 and Alexa 488; Invitrogen) for 1 hr at RT in PBS with 5% NGS. After rinsing with PBS, sections were mounted with Vectashield mounting medium containing DAPI (Vector Laboratories) and analyzed on a fluorescent microscope (Nikon E600).

Odajima J., Saini S., Jung P., Ndassa-Colday Y., Ficaro S., Geng Y., Marco E., Michowski W., Wang Y.E., DeCaprio J.A., Litovchick L., Marto J, & Sicinski P. (2016). Proteomic Landscape of Tissue-Specific Cyclin E Functions in Vivo. PLoS Genetics, 12(11), e1006429.

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Immunofluorescence Staining of Frozen Skin Lesions

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Frozen ENL skin lesion sections assays were performed in a Leica LM3000 cryostat, fixed in acetone, and hydrated in 0.01 M PBS. Unspecific binding sites were blocked with 10% Normal Goat Serum (NGS; Sigma-Aldrich) and 5% Fetal Calf Serum (FCS, GIBCO, Life Technologies) in 0.01 M PBS for 1 h at room temperature. Mouse IgG1 anti-human CD64 (1:50; Biolegend, 305002) and rabbit IgG anti-human MPO (1:100; Santa Cruz Biotechnology, SC-16128-R) and their respective isotypes were diluted in 1% Bovine Serum Albumin (BSA, Sigma-Aldrich), 1% NGS in 0.01 M PBS and incubated at 4°C overnight. Tissue sections were washed 3 times and incubated with Alexa Fluor 532 goat anti-mouse IgG (1:1000, ThermoFisher Scientific, A-11002) and Alexa Fluor 633 goat anti-rabbit IgG secondary antibodies (1:1000, ThermoFisher Scientific, A-21070) for 1:30 h at room temperature. The nuclei were stained with 4′-6-diamidino-2-phenylindole (DAPI; 1:10000, Molecular Probes, D1306), and slides were mounted with VECTASHIELD Mounting Medium (Vector Laboratories, H-1000). Tissues were imaged using an Axio Observer.Z1 fluorescence microscope equipped with a Colibri.2 illumination system (Carl Zeiss, Oberkochen, Germany) and the EC Plan-Neofluar 20x/0.50 objective and Plan-Apochromat 63x/1.3 oil objective. Images were acquired with a digital camera AxioCam HRm and AxioVision Rel. 4.6 software (Carl Zeiss).

Schmitz V., Prata R.B., Barbosa M.G., Mendes M.A., Brandão S.S., Amadeu T.P., Rodrigues L.S., Ferreira H., Costa F.D., dos Santos J.B., Pacheco F.D., Machado A.D., Nery J.A., Hacker M.D., Sales A.M., Pinheiro R.O, & Sarno E.N. (2016). Expression of CD64 on Circulating Neutrophils Favoring Systemic Inflammatory Status in Erythema Nodosum Leprosum. PLoS Neglected Tropical Diseases, 10(8), e0004955.

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Immunohistochemical Analysis of Spinal Cord

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Animals (n = 5/6 animals per group) were perfused through the heart with ice-cold saline, followed by a solution of 4 % paraformaldehyde in 0.1 M phosphate buffer, pH 7.4, via the ascending aorta. The spinal cords were then removed, and cryoprotected by immersion in 20 % sucrose followed by 30 % sucrose in phosphate buffer. The spinal cords were sectioned transversely at 20 μm on a cryostat (CM 1850; Leica, Wetzlar, Germany) and collected on gelatin-coated slides.
For immunohistochemistry, sections were washed three times with PBS with 0.3 % Triton X-100, incubated with 5 % normal goat serum (NGS; Sigma) in PBS for 30 minutes, and then incubated with the primary antibodies monoclonal mouse anti-NeuN (1:200, #MAB 377; Chemicon, Temecula, CA, USA) and polyclonal rabbit anti-Iba1 (1:400, #019-19741; Wako, Richmond, VA, USA) overnight at 4 °C. After the primary incubations, the appropriate secondary Cy3®-conjugated or Alexa® 488-conjugated antibodies (Jackson ImmunoResearch, West Grove, PA, USA and Invitrogen Inc., Carlsbad, CA, respectively) were used. In some cases, the sections were incubated with TO-PRO®-3 (1:1000; Invitrogen) for nuclei staining. Sections were mounted with VectaShield® (Vector, Burlingame, CA, USA) and analyzed using confocal microscopes (LSM 510 META or LSM 510 META NLO; Zeiss GmbH, Oberkochen, Germany).

Gubert F., Decotelli A.B., Bonacossa-Pereira I., Figueiredo F.R., Zaverucha-do-Valle C., Tovar-Moll F., Hoffmann L., Urmenyi T.P., Santiago M.F, & Mendez-Otero R. (2016). Intraspinal bone-marrow cell therapy at pre- and symptomatic phases in a mouse model of amyotrophic lateral sclerosis. Stem Cell Research & Therapy, 7, 41.

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