Mab3418x

Manufactured by Merck Group

Sourced in United States

MAB3418X is a laboratory equipment product manufactured by Merck Group. It is a versatile tool designed for a range of scientific applications. The core function of MAB3418X is to provide accurate and reliable measurements, but a detailed description while maintaining an unbiased and factual approach is not available.

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Lab products found in correlation

Lsm 880, by Zeiss (2 mentions) Ulex europaeus agglutinin 1, by Vector Laboratories (1 mentions) Anti cd31 antibody, by BioLegend (1 mentions) Alexa fluor 488, by Merck Group (1 mentions) Ab150075, by Abcam (1 mentions) P24g 1.0 10 f, by Merck Group (1 mentions) P6407, by Merck Group (1 mentions) D1306, by Thermo Fisher Scientific (1 mentions)

5 protocols using mab3418x

Immunofluorescence Staining of Spheroids

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For immunofluorescence staining, spheroids and cells were fixed in 4% (wt/vol) paraformaldehyde in phosphate‐buffered saline (PBS) for 15 min, permeabilized with 0.1% (vol/vol) Triton X‐100 in PBS for 30 min, and blocked in 3% (wt/vol) bovine serum albumin in PBS for 2 h at room temperature. Endothelial cells were marked using Ulex europaeus agglutinin 1 (Vector Laboratories) or anti‐CD31 antibody (catalog no. 303111; Biolegend). The neural spheroids were stained with Alexa Fluor 488‐conjugated anti‐olig2 antibody (MABN50A4; Sigma‐Aldrich), Alexa Fluor 594‐ conjugated anti‐SOX2 antibody (656106; Biolegend), Alexa Fluor‐488 conjugated anti‐neurofilament H antibody (801706; Biolegend), Alexa Fluor 594‐conjugated anti‐GFAP antibody (837509; Biolegend), Alexa Fluor 488‐conjugated anti‐β‐tubulin Class III antibody (560338; BD Bioscience), and Alexa Fluor 488‐conjugated anti‐MAP2 antibody (MAB3418X; Sigma‐Aldrich).

Shin N., Kim Y., Ko J., Choi S.W., Hyung S., Lee S., Park S., Song J., Jeon N.L, & Kang K. (2021). Vascularization of iNSC spheroid in a 3D spheroid‐on‐a‐chip platform enhances neural maturation. Biotechnology and Bioengineering, 119(2), 566-574.

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Quantifying Traumatic Brain Tissue Loss

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MAP2 staining and quantitation of traumatic brain tissue loss were performed as described previously [11 ]. Five equally spaced brain slices were used: +1.54 to − 3.46 from bregma, with 1000-μm interval. Coronal brain sections were stained with Alexa Fluor® 488-conjugated MAP2 antibody (1:200, MAB3418X, Sigma-Millipore, Burlington, MA, USA) and captured with an image scanner. The brain tissue loss was quantified with ImageJ software and calculated with the following formula: VC − VL (VC is the contralateral hemisphere volume; VL is the volume of residual tissue in the ipsilateral hemisphere).

Liu N., Han J., Li Y., Jiang Y., Shi S.X., Lok J., Whalen M., Dumont A.S, & Wang X. (2021). Recombinant annexin A2 inhibits peripheral leukocyte activation and brain infiltration after traumatic brain injury. Journal of Neuroinflammation, 18, 173.

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Quantification of Traumatic Brain Tissue Loss

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Quantitation of traumatic brain tissue loss was performed as described previously [39 (link)]. Briefly, six equally-spaced slices from bregma: +1.54 to −3.46, with 1000 μm interval. Coronal brain sections were stained with Alexa Fluor^® 488 conjugated MAP2 antibody (1:200, MAB3418X, Sigma-Millipore, Burlington, MA, USA), and the brain tissue loss was quantified with Image J software and calculated with the following formula: V_C–V_L (V_C is the volume of the contralateral hemisphere, V_L is the volume of residual tissue in the ipsilateral hemisphere).

Liu N., Jiang Y., Chung J.Y., Li Y., Yu Z., Kim J.W., Lok J.M., Whalen M.J, & Wang X. (2019). Annexin A2 Deficiency Exacerbates Neuroinflammation and Long-Term Neurological Deficits after Traumatic Brain Injury in Mice. International Journal of Molecular Sciences, 20(24), 6125.

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Quantifying Traumatic Brain Tissue Loss

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At 28 days after neurobehavioral tests, mice were deeply anesthetized and sacrificed for quantitation of traumatic brain tissue loss by following a standard method as we previously described (Liu et al., 2019 (link)). Briefly, mice were anesthetized and transcardially perfused with chilled (4°C) PBS, pH7.4, followed by 4% paraformaldehyde in 0.1M PBS. Frozen brains were sectioned with a microtome, and six equally-spaced slices (50-μm-thick) from bregma: +1.54 to −3.46, with 1000 μm interval. Coronal brain sections were stained with Alexa Fluor® 488 conjugated microtubule-associated protein 2 (MAP2) antibody (1:200, MAB3418X, Sigma-Millipore, Burlington, MA, United States), and the images of brain sections were captured with an image scanner. The brain tissue loss was quantified with a standard computer-assisted image analysis program (Image J), and calculated with the following formula: Brain tissue loss = contralateral hemisphere volume- ipsilateral hemisphere volume.

Cheng C., Wang X., Jiang Y., Li Y., Liao Z., Li W., Yu Z., Whalen M.J., Lok J., Dumont A.S., Liu N, & Wang X. (2021). Recombinant Annexin A2 Administration Improves Neurological Outcomes After Traumatic Brain Injury in Mice. Frontiers in Pharmacology, 12, 708469.

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Immunostaining of Neurite Markers in Cell Cultures

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Cells were grown on No. 1.0 glass bottom culture dishes (MatTek P24G-1.0–10-F) pretreated with poly-d-lysine (Sigma P6407) following the protocol described above for maintenance or differentiation conditions. At predetermined time, cells were washed with PBS, followed by a 30-min fixation period in 4% paraformaldehyde (PFA). Removal of PFA was followed by 3 × PBS wash, and a 1-h incubation in blocking buffer (3% FBS, 0.05% Triton X, PBS). Primary antibodies included NF200 (Abcam ab8135) and conjugated-Map2 (Sigma MAB3418X). Primary antibodies were diluted at a 1:1000 ratio in blocking buffer, added to cells, and incubated overnight at 4 °C in the dark. Cells were then washed 3 × in PBS, and incubated overnight at 4 °C in the dark after the addition of a secondary antibody (Abcam ab150075) diluted at 1:1000 in blocking buffer. Cells were washed 3 × in PBS and DAPI (Thermo D1306) was added for 1 min to stain nuclei. DAPI was removed and each well received ProLong Glass Antifade Mountant (Thermo P36982) per manufacturer’s instructions for imaging. Cells were imaged using a confocal microscope (Carl Zeiss LSM880) and three biological replicates per condition were analyzed at 20 × magnification. Five images were taken at randomized locations within tissue culture wells (n = 3), and representative images are used.

Wamil A.W., Wamil B.D, & Hellerqvist C.G. (1998). CM101-mediated recovery of walking ability in adult mice paralyzed by spinal cord injury. Proceedings of the National Academy of Sciences of the United States of America, 95(22), 13188-13193.

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