Abn78a4

Manufactured by Merck Group

Sourced in United States

The ABN78A4 is a laboratory equipment designed for general scientific applications. It serves as a versatile tool for researchers and technicians in various fields. The core function of this product is to provide a reliable and efficient platform for conducting a range of laboratory experiments and analysis.

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Lab products found in correlation

Ab183830, by Abcam (1 mentions) Hb050 inverted microscope system, by Zeiss (1 mentions) Olig2, by Abcam (1 mentions) Alexa fluor 488 goat anti, by Thermo Fisher Scientific (1 mentions) Alexa fluor 594 goat anti mouse igg, by Thermo Fisher Scientific (1 mentions) Neuronal nuclei (neun), by Merck Group (1 mentions) Clsm780 microscope, by Zeiss (1 mentions) Dapi fluoromount g, by Southern Biotech (1 mentions) Goat serum, by Southern Biotech (1 mentions) Sm2010r, by Leica (1 mentions) 4 6 diamidino 2 phenylindole fluoromount g, by Southern Biotech (1 mentions) Ab73593, by Abcam (1 mentions) Alexa fluor 488, by Thermo Fisher Scientific (1 mentions) Cm3050s cryostat, by Leica (1 mentions) Slowfade gold antifade mountant, by Thermo Fisher Scientific (1 mentions) Oct media, by Thermo Fisher Scientific (1 mentions) Ix71 inverted fluorescence microscope, by Olympus (1 mentions)

4 protocols using abn78a4

Immunofluorescence Staining of Brain and Cell Samples

Cited 1 time

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Immunofluorescence staining was performed according to our previously published protocols [41 (link), 42 (link)]. Briefly, the brain sections (6 μm thick) were incubated with primary antibodies against ALX/FPR2 (1:200, cat. no. ab203129, Abcam), GFAP (1:200, cat. no. MAB3402X, EMD Millipore, USA), Iba-1 (1:200, RRID: AB_2224402), and NeuN (1:200, cat. no. ABN78A4, EMD Millipore, USA) overnight at 4 °C. Then, the brain sections were incubated with the corresponding secondary antibodies conjugated with Alexa Fluor 488 and/or Alexa Fluor 594 (Jackson ImmunoResearch Incorporation, West Grove, PA, USA) for 1 h at room temperature, after which a ZEISS HB050 inverted microscope system was used for fluorescence detection.
For in vitro cell staining, primary antibodies against Iba-1 (1:200, RRID: AB_2224402) and p65 (1:200, 1:1000, cat. no. D14E12, CST) were used for microglia staining, while a primary antibody against MAP2 (1:500, cat. no. ab183830, Abcam) was used to stain neurons. The protocols and the secondary antibodies used for in vitro cell staining were the same as those used to stain brain sections.

Liu G.J., Tao T., Wang H., Zhou Y., Gao X., Gao Y.Y., Hang C.H, & Li W. (2020). Functions of resolvin D1-ALX/FPR2 receptor interaction in the hemoglobin-induced microglial inflammatory response and neuronal injury. Journal of Neuroinflammation, 17, 239.

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Immunohistochemical Profiling of Prefrontal Cortex

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Prefrontal tissue sections from a naive adult rhesus monkey were processed for immunohistochemistry with NeuN (Millipore, 1:1,000) and Olig2 (Abcam, 1:2,000) antibodies, in conjunction with Alexa Fluor 488 goat anti-rabbit and Alexa Fluor 594 goat anti-mouse IgGs (Invitrogen), using standard procedures. In a second, independent set of immunohistochemistry experiments, small tissue blocks (surface area <0.25 cm²) were dissected from the dorsolateral prefrontal cortex (Area9/46), immersion-fixed in 4 % paraformaldehyde solution for 15–18 hours, dehydrated in 70 %, 90 % and 100 % ethanol (3 × 30 min each) followed by xylene immersion (3 × 20 min) and paraffin-embedded using a Tissue TEK embedding center. Paraffin embedded tissue blocks were cut in 8 µm thick sections and mounted on slides for immunohistochemistry. Sections of three clozapine-exposed animals with elevated proportions of NeuN⁺ nuclei in white matter were de-paraffinized using xylazine and ethanol. The sections were rehydrated with water, permeabilized with 0.1 % Triton X-100, followed by NeuN staining (1:100, ABN78A4, EMD Millipore). After mounting the tissue with DAPI Fluoromount-G (0100-200, SouthernBiotech), images of NeuN⁺ subcortical white matter neurons were taken using a Carl Zeiss CLSM780 microscope. Image processing was done with ImageJ (NIH).

Halene T.B., Kozlenkov A., Jiang Y., Mitchell A., Javidfar B., Dincer A., Park R., Wiseman J., Croxson P., Giannaris E.L., Hof P.R., Roussos P., Dracheva S., Hemby S.E, & Akbarian S. (2016). NeuN+ Neuronal Nuclei in Non-Human Primate Prefrontal Cortex and Subcortical White Matter After Clozapine Exposure. Schizophrenia research, 170(0), 235-244.

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Perfusion-Fixed Brain Sectioning for Immunostaining

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Mice were anaesthetized with a terminal i.p. injection of a ketamine/xylazine mixture (IP: 200 and 30 mg kg⁻¹, respectively) 48 h after HSV injection. Intracardial perfusion was performed with 100 ml of 10% sucrose followed by 200 ml of 4% paraformaldehyde in PBS. Brains were removed and placed in 4% paraformaldehyde overnight at 4 °C, followed by incubation in 30% sucrose until isotonic. Brains were cut on a freezing microtome (Leica SM2010 R) into coronal sections (60 μm) and permeabilized and blocked with 0.1% Triton X-100 and 10% goat serum (Southern Biotech), respectively. Sections were incubated with NeuN-488 (1:500, EMD Millipore, ABN78A4) for 2 h, followed by a wash with PBS. Mounting was done using 4,6-diamidino-2-phenylindole Fluoromount-G (SouthernBiotech, 0100-20). Images were taken with a Carl Zeiss CLSM780 microscope and processed using ImageJ (NIH).

Mitchell A.C., Javidfar B., Bicks L.K., Neve R., Garbett K., Lander S.S., Mirnics K., Morishita H., Wood M.A., Jiang Y., Gaisler-Salomon I, & Akbarian S. (2016). Longitudinal assessment of neuronal 3D genomes in mouse prefrontal cortex. Nature Communications, 7, 12743.

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Immunostaining of ZIKV-infected Tissues

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Tissues were fixed in 4% PFA, embedded in OCT media (ThermoFisher, Waltham, MA, USA), and stored at −80 °C until sectioned into 10 µm cryosections using a Leica CM3050-S cryostat (Leica Biosystems, Buffalo Grove, IL, USA), and sections were stored at −80 °C until immunostaining. Sections were immunostained for ZIKV using a mouse-anti-ZIKV Envelope (E) protein primary antibody (FL0006, Kerafast, Boston, MA, USA), and visualized with a donkey-anti-mouse-AlexaFluor 488 secondary antibody (ab150101, Abcam, Cambridge, MA, USA). Sensory neurons were immunostained with anti-PGP9.5 (NB300-675, Novus Biologicals, Centennial, CO, USA) and satellite glial cells with anti-glutamine synthetase (ab73593, Abcam, Cambridge MA, USA) antibodies, followed by species-specific secondary antibodies conjugated to AlexaFluor 488 (Fisher Scientific, Hamptom, NH, USA). CNS neurons were immunostained with anti-NeuN conjugated to AlexaFluor 488 (ABN78A4, EMD Millipore, Burlington, MA, USA). Slides were treated with SlowFade Gold Antifade Mountant (ThermoFisher, Waltham, MA, USA) before cover slipping. Slides were visualized and imaged using an IX71 inverted fluorescence microscope (Olympus Life Sciences, Waltham, MA, USA).

Saver A.E., Crawford S.A., Joyce J.D, & Bertke A.S. (2019). Route of Infection Influences Zika Virus Shedding in a Guinea Pig Model. Cells, 8(11), 1437.

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