Mkb 2213 1

Manufactured by Vector Laboratories

The MKB-2213-1 is a laboratory instrument designed for performing various analytical techniques. It is a multi-functional device that can be utilized for a range of applications within a research or testing environment.

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Lab products found in correlation

Anti apoe, by Merck Group (1 mentions) Anti map2, by Merck Group (1 mentions) Upright microscope, by Nikon (1 mentions) One step hrp polymer, by Abcam (1 mentions) K405 50, by Abcam (1 mentions)

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MKB-2213 (18 citations)

3 protocols using mkb 2213 1

Quantification of Amyloid-Beta Brain Deposition

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The right hemisphere was collected and fixed either by immersion or perfusion. Then the brains were post-fixed in 4% PFA for 24 h and stored in 30% sucrose for at least 24 h. Tissue was stored at +4°C in cryoprotectant buffer (Sucrose 150 g, 0.1 M PB 200 mL, Ethylene glycol 150 mL, add more 0.1 M PB to reach a final volume of 500 mL) and shipped to PsychoGenics (Czechia). A systematic random set of 5 sagittal sections from 5 different mediolateral levels per mouse was labeled for Aβ, using an antibody to residues 1–4 of Aβ (MOAB2, Abcam 126649, dilution 1/1,000). Sections were counterstained with DAPI to visualize nuclei. Antibody binding was visualized using a highly cross-absorbed fluorescently labeled secondary antibody (Abcam 150106, dilution 1/500). All antibodies were diluted in antibody diluent (Dako, S 302283–2), and unspecific endogenous IgG binding was blocked with M.O.M. (Vector MKB 2213-1) before primary incubation. Entire brain sections were imaged on a Zeiss AxioScan.Z1 slide scanner microscope. Quantifications using the stored ROIs were done using Image Pro Premier software (v 9.1 or higher).

Overk C., Fiorini E., Babolin C., Vukicevic M., Morici C., Madani R., Eligert V., Kosco-Vilbois M., Roberts A., Becker A., Pfeifer A, & Mobley W.C. (2023). Modeling Alzheimer’s disease related phenotypes in the Ts65Dn mouse: impact of age on Aβ, Tau, pTau, NfL, and behavior. Frontiers in Neuroscience, 17, 1202208.

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Immunostaining of Primary Neurons

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Primary neurons on coverslips were washed 10min in dPBS, then permeabilized in dPBS + 0.1% Triton-X for 5min at RT. The neurons were then blocked in 10% normal donkey serum + 0.5% Triton in PBS for 1hr, and further blocked with mouse-on-mouse (MOM) blocking buffer (Vector Labs, MKB-2213-1) at 1 drop Mouse IgG block buffer per 4ml PBS. Primary antibodies were diluted in MOM Antibody dilution buffer (80ul protein concentrate to 1ml PBS), and cells were incubated in primary antibody solution overnight at 4°C (anti-ApoE 1:5000, Millipore 178479; anti-MHC-I (OX18) 1:50, Santa Cruz Biotechnologies sc-53074; anti-MAP2 1:500, Millipore ab5622; anti-p-Tau (PHF1) 1:300, from Peter Davis). The next day, cells were washed 3x (15min, 10min, 5min) in PBS with 0.1% Triton-X. Secondary antibodies were diluted in MOM Antibody Dilution Buffer (1:1500), and incubated with the cells for 1hr at RT. Cells were again washed 3x (15min, 10min, 5min) in PBS with 0.1% Triton-X. Coverslips were then mounted to microscope slides (Vectashield, Prolong Gold).

Zalocusky K.A., Najm R., Taubes A.L., Hao Y., Yoon S.Y., Koutsodendris N., Nelson M.R., Rao A., Bennett D.A., Bant J., Amornkul D.E., Xu Q., An A., Cisne-Thomson O, & Huang Y. (2021). Neuronal ApoE Upregulates MHC-I Expression to Drive Selective Neurodegeneration in Alzheimer’s Disease. Nature neuroscience, 24(6), 786-798.

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Immunohistochemical Analysis of Autophagy and Macrophages

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Immunohistochemistry for detection of the LC3 protein and the macrophage marker F4/80 was carried out using standard protocols. Briefly, unstained sections of lungs or tumors were deparaffinized and rehydrated, and then incubated in the peroxidase blocking reagent (BioVision cat #K405-50). Antigen retrieval was perfomed by boiling the sections in sodium citrate for 20 min. To decrease background staining, the slides were incubated for 1 h in the mouse on mouse blocking reagent (Vector Labs MKB-2213-1), followed by overnight incubation with the primary antibodies (LC3B Cell Signaling 3868S rabbit polyclonal, 1:300; or F4/80 Cell Signaling 70076S rabbit monoclonal, 1:200). Next day, the slides were washed and incubated with One-Step HRP polymer (BioVision cat #K405-50) for 30 min at RT. The slides were then washed several times and then incubated with the DAB chromogen for 10 min at RT, followed by several washing steps and quick counterstain with Hematoxylin. Slides were then mounted and visualized under the upright microscope (Nikon).

Zarea A.A., Perez G.I., Broadbent D., Dolgikh B., Bernard M.P., Withrow A., McGill A., Toomajian V., Thampy L.K., Harkema J.R., Walker J.R., Kirkland T.A., Bachmann M.H., Schmidt J.C., & Kanada M. (2021). In vitro and in vivo analysis of microvesicle-mediated metastasis using a bright, red-shifted bioluminescent reporter protein of extracellular vesicles.

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