Mab1281

Manufactured by Merck Group

Sourced in United States, United Kingdom, Germany

MAB1281 is a laboratory product manufactured by Merck Group. It is a monoclonal antibody used in research and diagnostic applications. The core function of MAB1281 is to serve as a detection and quantification tool for specific target analytes in various sample types.

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Lab products found in correlation

Tween 20, by Merck Group (3 mentions) Alexa fluor 488, by Thermo Fisher Scientific (3 mentions) Ab7349, by Abcam (2 mentions) Tissue tek oct, by Sakura Finetek (2 mentions) Smi 21, by Fortrea (2 mentions) Lsm 700, by Zeiss (2 mentions) Normal goat serum (ngs), by Merck Group (2 mentions) Normal goat serum (ngs), by Thermo Fisher Scientific (2 mentions) Hoechst 33258 bisbenzimideh 33258 trihydrochloride, by Merck Group (2 mentions) Fluoromount, by Merck Group (2 mentions) Ab28364, by Abcam (2 mentions) Cy3 fitc conjugated anti mouse rabbit anti igg, by Thermo Fisher Scientific (2 mentions) Picric acid, by Merck Group (2 mentions) Alexa fluor 568, by Thermo Fisher Scientific (2 mentions) Alexa fluor 594, by Thermo Fisher Scientific (2 mentions) Scid mice, by Jackson ImmunoResearch (2 mentions) Matrigel, by Merck Group (2 mentions) Matrigel, by BD (2 mentions) Lionheart fx automated live cell imager, by Agilent Technologies (1 mentions) Ab18723, by Abcam (1 mentions)

51 protocols using mab1281

Hemangioblast-Derived Endothelial Cell Assay

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Example 4

Hemangioblast-derived endothelial cells or pure hemangioblasts (1×10⁶cells) derived from H1-GFP human ES cells were purified and resuspended in 700 μl of Matrigel (BD Biosciences™). These cells were injected subcutaneously in the dorsal region of 4-week-old SCID mice (Jackson Laboratory™). Five weeks after injection, the Matrigel plugs were removed from the animals and frozen sections were prepared from the plugs. Cross sections of the Matrigel plugs were fixed, and stained by standard hematoxylin and eosin (H&E) staining, as well as immunohistochemistry using an anti-human specific nuclei antibody (MAB1281, Chemicon™) and human specific anti-vWF antibody, followed by fluorescent dye labeled corresponding secondary antibodies (goat anti-mouse IgG-Texas Red secondary antibody for MAB1281). FIG. 10 demonstrates vessels in cross sections of Matrigel plugs with H&E staining; and FIG. 11 demonstrates that the vessels in cross sections of Matrigel plugs are cells derived from human hemangioblasts, which are positive for human specific nuclei antibody.

US09938500B2. Hemangio-colony forming cells (2018-04-10). Astellas Institute for Regenerative Medicine [US]. Inventors: Robert Lanza [US], Shi-Jiang Lu [US].

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Hemangioblast-Derived Endothelial Cell Transplantation

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Example 4

US11566228B2. Hemangio-colony forming cells (2023-01-31). Astellas Institute for Regenerative Medicine [US]. Inventors: Robert Lanza [US], Shi-Jiang Lu [US].

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Quantitative Immunofluorescence Microscopy

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Mouse anti-human nuclei monoclonal antibody (MAB1281 Millipore Merck) was used for immunofluorescence staining procedures, according to the manufacturer’s instructions.
Ten randomly selected fields per each slide were examined, and the areas of interest were interactively selected by two independent observers, using proper optical threshold and microscope filter combinations. Computer morphometry was performed using Lionheart FX Automated Live Cell Imager (BioTek VT, USA). The system included a digital color camera and software package for immunofluorescence evaluation.

Zilberman-Itskovich S., Abu-Hamad R., Zarura R., Sova M., Hachmo Y., Stark M., Neuman S., Slavin S, & Efrati S. (2019). Human mesenchymal stromal cells ameliorate complement induced inflammatory cascade and improve renal functions in a rat model of ischemia-reperfusion induced acute kidney injury. PLoS ONE, 14(9), e0222354.

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Immunohistochemical Labeling of Human Nuclei

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Sections were fixed in formaldehyde. A monoclonal mouse antibody was used for labeling human nuclei (anti-human nuclei, HuNu, MAB 1281, 1:200, EMD Millipore, UK). Sections were incubated overnight at 4 °C. Biotin block solution (Vector Laboratories Inc., Burlingame, CA, USA) was added to the primary antibody dilution. Furthermore, antibody conjugates were adsorbed with normal rabbit serum to minimize cross-reactivity. An avidin-conjugated secondary antibody was used. For analysis of adverse effects of the transplantation, such as thromboembolic complications, hematoxylin and eosin staining was performed according to Mayer's protocol.¹⁹

Arnberg F., Lundberg J., Kenne E., Jaff N., Müller P., Nava S., Kaipe H., Ringdén O, & Holmin S. (2014). Superselective intra-arterial umbilical cord blood administration to BM in experimental animals. Bone Marrow Transplantation, 49(12), 1486-1491.

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Cellular and Tissue Immunofluorescence Staining

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The cellular immunofluorescence staining for Lamin B1 (1:100, CL594-66095, Proteintech, China) was performed to detect cell senescence. For tissue immunofluorescence staining, after blocking, the frozen sections were incubated with primary antibodies against NeuN (1:1000, ab177487, Abcam, USA), Aβ (mouse anti-Aβ (1:200, SIG-39320, Covance), DCX (1:100, ab18723, Abcam, USA), MAB1281 (1:100, MAB1281, Merck, USA), GFAP (1:500, 16825-1-AP, Proteintech, China) and Iba1 (1:100, 10904-1-AP, Proteintech, China) overnight at 4 °C, and then incubated with Cy3 or FITC-conjugated anti-mouse or ant-rabbit IgG (1:200, AB0122, Abways, China) for 2 h. For EdU/DCX, EdU/NeuN, or EdU/Nestin double immunofluorescence staining, EdU solution (5 mg/kg) was intraperitoneally injected daily for three consecutive days before the mice were sacrificed, according to the instructions of the Cell-Light EdU Apollo567Kit (RiboBio, China). After Apollo staining, the slices were respectively incubated with DCX, NeuN or Nestin antibody (1:1000, 19483-1-AP, Proteintech, USA) overnight at 4 °C and incubated with FITC-conjugated IgG antibody (1:500, SA00003-2, Proteintech, USA) for 2 h. After DAPI (1:100) counterstaining, the sections were examined under a microscope (Leica, German), and the positive cells were analyzed using Image J software (Bethesda, USA).

Ma S., Zhou X., Wang Y., Li Z., Wang Y., Shi J, & Guan F. (2022). MG53 protein rejuvenates hUC-MSCs and facilitates their therapeutic effects in AD mice by activating Nrf2 signaling pathway. Redox Biology, 53, 102325.

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Tracking Transplanted Cells in Spinal Cord

Cited 1 time

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The pigs were euthanized 21 days after transplantation. Transcardiac perfusion with 0.9% NaCl solution followed by 4% paraformaldehyde was performed. The fixed spinal cord was excised and frozen. The cord was sectioned axially at 50 μm intervals and stained with Prussian Blue (PB) reagent for microscopic Iron and counter-stained with Eosin. Immunohistochemical staining for detection of grafted human cells using a primary mouse monoclonal anti-human nucleus (HuNu) antibody (MAB1281; EMD Millipore; 1/250) and detection of grafted pig cells using a primary mouse monoclonal anti-GFP antibody (AB3080; EMD Millipore; 1/250) was performed on every 6^th section with cresyl violet background stain. Images were captured using a Nikon E400 microscope supplied with NIS-Elements imaging software (Nikon Instruments, Inc.). The current study was not designed to investigate the therapeutic efficacy or biological properties of transplanted cell grafts, which has been established in previous studies[29 (link)–31 (link)].

Lamanna J.J., Urquia L.N., Hurtig C.V., Gutierrez J., Anderson C., Piferi P., Federici T., Oshinski J.N, & Boulis N.M. (2017). Magnetic Resonance Imaging-Guided Transplantation of Neural Stem Cells into the Porcine Spinal Cord. Stereotactic and functional neurosurgery, 95(1), 60-68.

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Nuclei Immunostaining and FISH Analysis

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Immunostaining for human cell nuclei was carried out by using anti-nuclei antibodies (MAB1281, Merck Millipore) and FISH analysis by using the CEP X SpectrumOrange/Y SpectrumGreen Direct Labeled Fluorescent DNA Probe Kit (Abbott Molecular), both following the manufacturer’s protocols.

Su L.J., Wu M.S., Hui Y.Y., Chang B.M., Pan L., Hsu P.C., Chen Y.T., Ho H.N., Huang Y.H., Ling T.Y., Hsu H.H, & Chang H.C. (2017). Fluorescent nanodiamonds enable quantitative tracking of human mesenchymal stem cells in miniature pigs. Scientific Reports, 7, 45607.

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Immunohistochemical Profiling of Cryopreserved Brains

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Brains were cryopreserved, embedded in OCT (Tissue-Tek OCT, Sakura Finetek, Torrance, CA) and sectioned at 20 μm, either sagittally or coronally, on a cryostat. Human cells were identified with mouse anti-human nuclei, clone 235–1 at 1:800 (MAB1281; EMD Millipore, Billerica, MA). Oligodendrocytes were labeled with myelin basic protein with rat anti-MBP at 1:25 (Ab7349; Abcam, Cambridge, MA), astrocytes with anti-human-specific GFAP (SMI 21 at 1:1000, Covance, Princeton, NJ), and axons with mouse anti-neurofilament at 1:5000 (SMI-311) or 1:1000 (SMI-312; Covance, Princeton, NJ). Alexa Fluor secondary antibodies, goat anti-mouse and anti-rat 488, 568, 594, and 647 were used at 1:400 (Life Technologies, Carlsbad, CA).

Osipovitch M., Asenjo-Martinez A., Mariani J.N., Cornwell A., Dhaliwal S., Zou L., Chandler-Militello D., Wang S., Li X., Benraiss S.J., Agate R., Lampp A., Benraiss A., Windrem M.S, & Goldman S.A. (2018). Human ESC-derived chimeric mouse models of Huntington disease reveal cell-intrinsic defects in glial progenitor cell differentiation. Cell stem cell, 24(1), 107-122.e7.

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Cryosectioning and Immunolabeling of Human Brains

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Brains were cryopreserved, embedded in OCT (Tissue-Tek OCT, Sakura Finetek, Torrance, CA) and sectioned at 20 μm, either sagittally or coronally, on a cryostat. Human cells were identified with mouse antihuman nuclei, clone 235-1 at 1:800 (MAB1281, EMD Millipore, Billerica, MA). Myelin basic protein was labeled with rat anti-MBP at 1:25 (Ab7349, Abcam, Cambridge, MA), oligodendrocyte progenitors with anti-human-specific PDGFRa (D13C6, XP® rabbit mAb 5241, 1:300, Cell Signaling Tech), oligodendrocytes with mouse anti-human-specific transferrin (clone HT1/13.6.3, 08691231, MP Biomedicals), astrocytes with anti-human-specific GFAP (SMI 21 at 1:1000, Covance, Princeton, NJ). Alexa Fluor secondary antibodies, goat anti- mouse, rat, and rabbit 488, 568, 594, and 647 were used at 1:400 (Life Technologies, Carlsbad, CA).

Windrem M.S., Osipovitch M., Liu Z., Bates J., Chandler-Militello D., Zou L., Munir J., Schanz S., McCoy K., Miller R.H., Wang S., Nedergaard M., Findling R.L., Tesar P.J, & Goldman S.A. (2017). Human iPSC glial mouse chimeras reveal glial contributions to schizophrenia. Cell stem cell, 21(2), 195-208.e6.

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Immunohistochemical Analysis of Brain Tissue

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After perfusion, brains were post-fixed for 24 h in 4% PFA and then cryopreserved in a 30% sucrose solution before being sectioned coronally on a freezing sledge microtome at a thickness of 35 μm in series of 1:8 or 1:12.
Immunohistochemistry was performed on free-floating sections and all washing steps were carried out with 0.1 M phosphate buffered saline with potassium (KPBS, pH 7.4). The sections were washed three times and then incubated in Tris-EDTA pH 8 for 30 min at 80 °C for antigen retrieval. After washing an additional three times, the sections were incubated in blocking solution (5% serum, 0.25% Triton X-100) for 1 h, before adding the primary antibody solution. Primary antibodies used were: mouse anti-HuNu (1:200, Merck Millipore, #MAB1281), rabbit anti-TH (1:1000, Merck Millipore, #AB152) and chicken anti-GFP (1:1000, Abcam, #ab13970). After incubation with primary antibodies overnight at room temperature, the sections were washed twice and incubated in blocking solution for 30–45 min. For fluorescent immunolabeling, the sections were then incubated with fluorophore-conjugated secondary antibodies (1:200, Jackson ImmunoResearch Laboratories) for 2 h at room temperature, washed three times and then mounted on gelatin-coated slides and cover-slipped with PVA-DABCO containing DAPI (1:1000).

Fiorenzano A., Birtele M., Wahlestedt J.N, & Parmar M. (2021). Evaluation of TH-Cre knock-in cell lines for detection and specific targeting of stem cell-derived dopaminergic neurons. Heliyon, 7(1), e06006.

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