M o m kit

Manufactured by Vector Laboratories

Sourced in United States, Canada, United Kingdom

The M.O.M. kit is a laboratory product designed for cell culture applications. It functions as a cell detachment solution, facilitating the separation of adherent cells from culture surfaces. The kit contains the necessary reagents and components to perform this cell dissociation process.

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Lab products found in correlation

Ph 6 sodium citrate buffer, by BioGenex (6 mentions) Dab kit, by Vector Laboratories (6 mentions) Monoclonal mouse anti human cd45, by Agilent Technologies (5 mentions) Hoechst 33342, by Thermo Fisher Scientific (5 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (5 mentions) Mouse anti brdu antibody, by BD (4 mentions) Bromodeoxyuridine (brdu), by Merck Group (4 mentions) Abc kit, by Vector Laboratories (4 mentions) Triton x 100, by Merck Group (4 mentions) Alexa fluor 488, by Thermo Fisher Scientific (4 mentions) Vectastain elite abc kit, by Vector Laboratories (3 mentions) Mouse anti β catenin, by BD (3 mentions) Immpact dab, by Vector Laboratories (3 mentions) Hoechst, by Merck Group (3 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (3 mentions) Lsm 780 confocal microscope, by Zeiss (3 mentions) Lsm 510, by Zeiss (2 mentions) Vectamount, by Vector Laboratories (2 mentions) Cryostat, by Leica (2 mentions) Tcs sp8 system, by Leica (2 mentions)

Spelling variants of this product

M.O.MTM Kit (2 citations) Vector M.O.M. peroxidase or fluoresceinImmunodetection Kits (2 citations)

126 protocols using m o m kit

Immunohistochemistry of Muscle Progenitors

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Antibodies for immunohistochemistry included: Pax7 (a gift from Chen-Min Fan, Carnegie Institute), Ki67 (BD Pharmagen), and MyoD (Santa Cruz). All immunostaining was perfomed with citrate-based antigen retrieval, and M.O.M. Kits (Vector Labs) were used for Pax7 staining.

Dodd R.D., Sachdeva M., Mito J.K., Eward W.C., Brigman B.E., Ma Y., Dodd L., Kim Y., Lev D, & Kirsch D.G. (2015). Myogenic transcription factors regulate pro-metastatic miR-182. Oncogene, 35(14), 1868-1875.

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Immunohistochemistry of Muscle Progenitors

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Immunohistochemical Analysis of Spinal Cord

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After sacrifice, mice were perfused (PBS then paraformaldehyde, PFA, 4%,) and lumbar spinal cord tissues were harvested, left in PFA (3 days) and embedded in paraffin. 5μm sections were deparaffinized, rehydrated, submerged in DAKO–TARGET solution (cat. S-1699, 20min) and incubated in 3% H₂O₂ (5min). Non-specific biotin/avidin (Vectorlabs, Burlingame, CA) and Fc/Fab staining was blocked using kits (cat. 015.000.008 Jackson ImmunoResearch). M.O.M kits (Vectorlabs, Burlingame, CA) were used to dilute primary mouse anti-mouse/human IDO mAb (E7, 1:50; Santa Cruz Biotechnology, cat. 365086), neurons (NeuN rabbit polyclonal, 1:500; Millipore cat. ABN78), microglia (IBA rabbit, 1:500; WAKO, cat. 019-19741) and astrocytes (GFAP rabbit monoclonal, 1:500; Cell Signaling, cat. 123895). After 60 min, sections were incubated with biotinylated anti-mouse IgG antibody from the M.O.M kit and Alexa fluor 488-conjugated AffiniPure F(ab′)2 fragment donkey anti-rabbit IgG (H+L) (cat. 711-546-152, Jackson ImmunoResearch) for 45 min. After washing, streptavidin conjugated with Alexa fluor 555 (1:400, Invitrogen Molecular probes, cat. S32355) was applied for 15 min. Slides were mounted in Fluor Save Reagent (Calbiochem, cat. 345789) and analyzed by confocal microscopy (Zeiss LSM 510).

Lemos H., Huang L., Chandler P.R., Mohamed E., Souza G.R., Li L., Pacholczyk G., Barber G.N., Hayakawa Y., Munn D.H, & Mellor A.L. (2014). Activation of the Stimulator of Interferon Genes (STING) adaptor attenuates experimental autoimmune encephalitis. Journal of immunology (Baltimore, Md. : 1950), 192(12), 5571-5578.

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Immunohistochemical Analysis of Tau in NSun2 KO Mouse Brain

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Brain embedded sections from 11-month-old NSun2 KO and controls (7 microns thick) were deparaffinized in Histo-Clear II (National Diagnostics, GA) and processed for immunohistochemistry using anti-mouse and rabbit tau antibodies (see below) according to the manufacturer’s protocol for mouse brain sections (MOM kit; Vector Labs, Cat # PK-2200). A 30 min incubation with 3% H2O2/10% methanol/0.25% Triton X-100 was used to block endogenous peroxidase activity. 3,3′-Diaminobenzidine was used as a peroxidase substrate (Vector DAB Substrate Kit for Peroxidase, Cat # SK-4100). Tissue sections were counterstained with hematoxylin (Vector Labs, Cat # H-3404) and mounted using VectaMount (Vector Labs, Cat # H-5000). Images were captured using an Olympus BX53 microscope with an Olympus camera DP-72 (Olympus Lifescience).

Kim Y.A., Siddiqui T., Blaze J., Cosacak M.I., Winters T., Kumar A., Tein E., Sproul A.A., Teich A.F., Bartolini F., Akbarian S., Kizil C., Hargus G, & Santa-Maria I. (2022). RNA methyltransferase NSun2 deficiency promotes neurodegeneration through epitranscriptomic regulation of tau phosphorylation. Acta Neuropathologica, 145(1), 29-48.

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Cryosectioning and Immunostaining of Frozen Muscle Tissue

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Frozen TA muscles embedded in OCT were cryosectioned using a Leica CM3050 S Cryostat. Tissue sections 10 μm thick were mounted onto Superfrost Plus glass microscope slides (Thermo Fisher Scientific) and kept at –20°C until immunostained. When thawed, the sections were fixed with ice-cold acetone for 10 min at –20°C. We employed the ‘mouse-on-mouse’ (MOM) kit (Vector Laboratories) to reduce non-specific antibody staining per the manufacturer’s specifications. Antibodies (key resource table) were used sequentially then slides were incubated with Hoechst block for 10 min. Following 2× for 5 min PBS washes, the slides were dried and coverslips mounted with Fluorogel. Fluorescent images were taken using a Leica DMR fluorescence microscope.

Guo D., Daman K., Chen J.J., Shi M.J., Yan J., Matijasevic Z., Rickard A.M., Bennett M.H., Kiselyov A., Zhou H., Bang A.G., Wagner K.R., Maehr R., King O.D., Hayward L.J, & Emerson CP J.r. (2022). iMyoblasts for ex vivo and in vivo investigations of human myogenesis and disease modeling. eLife, 11, e70341.

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Myosin Expression in Myoblasts

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Myoblasts were grown on glass coverslips coated with rat tail collagen and then treated with either the HERG-encoded or the control virus and allowed to terminally differentiate. These were then immunostained for myosin using the DSHB antibody recognizing myosin and a mouse on mouse (M.O.M.) Kit (Vector Labs, Inc.; Burlingame, CA) per manufacturer’s instructions. The coverslips were then mounted to slides with a mounting substance containing DAPI, and images were acquired using a Leica DM4500 microscope with a Leica DFC 340FX camera. The nuclei of myosin-positive cells were counted in three fields from ten slides (five treated with HERG-encoded virus and five treated with control virus).

Whitmore C., Pratt E.P., Anderson L., Bradley K., Latour S.M., Hashmi M.N., Urazaev A.K., Weilbaecher R., Davie J.K., Wang W.H., Hockerman G.H, & Pond A.L. (2020). The ERG1a potassium channel increases basal intracellular calcium concentration and calpain activity in skeletal muscle cells. Skeletal Muscle, 10, 1.

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Immunohistochemistry of Muscle Sections

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Immunohistochemistry was carried out on 10 μm frozen sections, as previously described [13 (link)]. Muscles to be assayed by immunohistochemistry were harvested and placed in OCT (Sakura Finetek, Torrance, CA, USA) then frozen in isopentane cooled by liquid nitrogen. Muscles were cross-sectioned (10 μm) and stained using the MOM kit (Vector Laboratories, Burlingame, CA, USA) and either the AEC Peroxidase Substrate Kit (Vector Laboratories) or using fluorophore-conjugated antibodies. Antibodies used were developmental myosin heavy chain (1:25, Leica Biosystems, Teban Gardens Crescent, Singapore), CD11b (1:50, BD PharMingen, San Diego, CA, USA), CD68 (1:50, BD PharMingen), and luciferase (1:25, Promega, Madison, WI, USA). Images were acquired using a compound microscope (Zeiss, Ontario, CA, USA), processed using AxioVision software (Zeiss), and displayed using Photoshop CS4 (Adobe, Mountain View, CA, USA).

Martinez L., Ermolova N.V., Ishikawa T.O., Stout D.B., Herschman H.R, & Spencer M.J. (2015). A reporter mouse for optical imaging of inflammation in mdx muscles. Skeletal Muscle, 5, 15.

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Immunohistochemistry and Immunocytochemistry for Muscle Stem Cells

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For immunohistochemistry, paraffin-embedded muscle sections were deparaffinized in xylene followed by rehydration. The slides were incubated at 95°C in sodium citrate buffer (10 mM Sodium Citrate, 0.05% Tween 20, pH 6.0) for 30 min followed by 20 min incubation at room temperature. Pax7-positive SCs were detected using a M.O.M kit from Vector Laboratories (Burlingame, CA) according to the manufacturer’s instruction. To stain nuclei, DAPI was simultaneously applied with the secondary antibodies at a concentration of 0.1%. Images were acquired using a Zeiss LSM 510 meta confocal microscope. Ten random fields per muscle section corresponding to approximately 1,000 fibers per muscle were imaged and counted. For immunocytochemistry, 3 day differentiated SCs were fixed in 4% paraformaldehyde at room temperature for 10 min followed by permeabilization in 0.2% triton X-100 for 3 min. Cells were blocked in 5% goat serum for 30 min at room temperature before applying anti-myosin primary antibody (MF20).

Shi H., Gatzke F., Molle J.M., Lee H.B., Helm E.T., Oldham J.J., Zhang L., Gerrard D.E, & Bennett A.M. (2015). Mice lacking MKP-1 and MKP-5 Reveal Hierarchical Regulation of Regenerative Myogenesis. Journal of stem cell and regenerative biology, 1(1), 1-7.

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Quantifying Cell Proliferation After Stroke

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The S-phase marker 5-bromo-2-deoxyuridine (BrdU) was used to label cells that underwent proliferation at the time of BrdU injection. BrdU (50 mg/kg body weight, Sigma-Aldrich) was injected intraperitoneally daily beginning 3 days after MCAO and continuing until 10 days after MCAO. Animals were subsequently deeply anesthetized and transcardially perfused with 0.9% NaCl followed by 4% paraformaldehyde in phosphate buffer. Brains were then sequentially cryoprotected in 20% and 30% sucrose, and frozen serial coronal brain sections (25 μm) were prepared on a cryostat (Leica, Bensheim, Germany). Sections were pretreated with 2N HCl for 1 hour at 37°C, followed by 0.1M boric acid (pH 8.5) for 10 minutes at room temperature, and then blocked with the M.O.M kit (Vector, Brulingame, CA, USA), followed by incubation with mouse anti-BrdU antibody (1:1000, BD bioscience, San Jose, CA, USA) for 1 hour at room temperature and overnight at 4°C. After a series of washes, sections were then incubated in 594 or 488-conjugated goat anti-mouse IgGs (1:1000, Jackson ImmunoResearch Laboratories, Inc. West Grove, PA, USA) for 1 hour at room temperature, rinsed, counterstained with DAPI for visualization of nuclei, and coverslipped for microscopic evaluation.

Cai M., Zhang W., Weng Z., Stetler R.A., Jiang X., Shi Y., Gao Y, & Chen J. (2017). Promoting Neurovascular Recovery in Aged Mice after Ischemic Stroke - Prophylactic Effect of Omega-3 Polyunsaturated Fatty Acids. Aging and Disease, 8(5), 531-545.

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Tissue Imaging and Immunostaining Protocol

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Tissues sections were first deparaffinized and subjected to antigen retrieval using a citrate buffer at 95 °C for 20 minutes as previously described^{7 (link)}. Tissue morphology was next assessed by hematoxylin and eosin (H&E) staining as previously described^{7 (link)}. Collagen deposition and localization were visualized by Trichrome as previously described^{7 (link)}. Immunostaining was performed by using antibodies of Col-1 (1310-01, SouthernBiotech), α-SMA (1A4, R&D systems) and KIF5A (ab118534, Abcam) as previously described^{7 (link)}. Briefly, mouse tissue sections were blocked using a proprietary blocking solution from a M.O.M. kit (Vector Laboratories). Primary antibodies were then incubated overnight at 4 °C in kit diluent. First antibodies were visualized with Alexa Fluor 488, 568, and 647 secondary antibody (Liftechnologies), and nuclei were stained with Hoechst 33342 (ThermoFisher Scientific). Tissues were mounted onto slides with ProLong Gold Antifade Reagent (Lifetechnologies). Tissue staining images were taken by ECLIPSE Ti (Nikon) for Trichrome staining and Leica TCS SP8 systems (Leica Microsystems) for immunofluorescence staining.

Kamata H., Tsukasaki Y., Sakai T., Ikebe R., Wang J., Jeffers A., Boren J., Owens S., Suzuki T., Higashihara M., Idell S., Tucker T.A, & Ikebe M. (2017). KIF5A transports collagen vesicles of myofibroblasts during pleural fibrosis. Scientific Reports, 7, 4556.

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