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Mhcii

Manufactured by Agilent Technologies
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The MHCII is a lab equipment product from Agilent Technologies. It is used for the detection and analysis of major histocompatibility complex class II (MHC-II) proteins. The MHCII device enables researchers to study the role of MHC-II molecules in immune responses and other biological processes.

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Lab products found in correlation

Glial fibrillary acidic protein (gfap), by Cell Signaling Technology (1 mentions) Ep1536y, by Abcam (1 mentions) Cd11b, by Abcam (1 mentions) Cd11b, by Agilent Technologies (1 mentions) Target retrieval solution s1699, by Agilent Technologies (1 mentions) Immpress reagent, by Vector Laboratories (1 mentions) Alexa fluor conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Southern Biotech (1 mentions) Bx60 microscope, by Olympus (1 mentions) Neuronal nuclei (neun), by Abcam (1 mentions) Abc peroxidase kit, by Vector Laboratories (1 mentions) Dab substrate kit, by Vector Laboratories (1 mentions)

6 protocols using mhcii

1

Immunohistochemical Analysis of Mouse Brain

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Mice were euthanized by intra-cardiac perfusion and brains were fixed in neutral buffered formalin. Paraffin-embedded brain sections were assessed with the following antibodies: GFAP (Cell Signaling, 1:1000), Iba1 (Wako, 1:1000); pSer129/81A αSyn (1:3000; [13 (link), 36 (link)]); EP1536Y (AbCam, 1:1000); p62 (Protein Tech; 1:1000), cd11b (AbCam; 1:250), MHCII (Novus; 1:50) and NeuN (Abcam; 1:500). For all antibodies except cd11b and MHCII, antigen retrieval was performed by steaming for 25 min in water. For cd11b and MHCII, antigen retrieval was done by steaming in Dako Target Retrieval Solution S1699 (modified citrate buffer, pH 6.1). Colorimetric slides were treated with ImmPress reagents (Vector Labs) and visualized with 3, 3’diaminobenzidine followed by hematoxylin counterstaining. Fluorimetric slides were visualized with Alexa Fluor conjugated secondary antibodies (Invitrogen) and counterstaining with 4’, 6-diamidino-2-phenylindole (DAPI; Southern Biotech). All colorimetric slides were scanned using the Aperio XT whole slide scanner while fluorescent slides were visualized using Olympus BX60 microscope with a color digital camera.
Koller E.J., Brooks M.M., Golde T.E., Giasson B.I, & Chakrabarty P. (2017). Inflammatory pre-conditioning restricts the seeded induction of α-synuclein pathology in wild type mice. Molecular Neurodegeneration, 12, 1.
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2

Immunohistochemical Profiling of Alzheimer's

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Antibodies used included 6E10 for Aβ amyloid plaques; Glial fibrillar acidic protein (GFAP) for astrocytes (Dako) and MHC-II (Dako) for microglia.
Snow A.D., Castillo G.M., Nguyen B.P., Choi P.Y., Cummings J.A., Cam J., Hu Q., Lake T., Pan W., Kastin A.J., Kirschner D.A., Wood S.G., Rockenstein E., Masliah E., Lorimer S., Tanzi R.E, & Larsen L. (2019). The Amazon rain forest plant Uncaria tomentosa (cat’s claw) and its specific proanthocyanidin constituents are potent inhibitors and reducers of both brain plaques and tangles. Scientific Reports, 9, 561.
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3

Reactive Microglia Labeling Protocol

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The protocol used for reactive microglia labelling was as that described for GFAP, with the exception of primary antibody. In this case, CD68 (1/500, 30 min RT; DAKO) and MHCII (1/200, 30 min RT; DAKO) were used.
Garcés M., Guijarro I.M., Ritchie D.L., Badiola J.J, & Monzón M. (2021). Novel Morphological Glial Alterations in the Spectrum of Prion Disease Types: A Focus on Common Findings. Pathogens, 10(5), 596.
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4

Immunohistochemical Analysis of HLA-DR

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Fixed tissue was available for 30 of 94 cases described in Table 1 that had frozen tissue. To increase our sample size, we acquired additional cases to provide sufficient cases with fixed tissue to measure HLA-DR by immunohistochemistry as described in Table 1. Tissue sections from the frontal cortex were sectioned by vibratome at 50 μm and stained by free-floating immunohistochemistry for HLA-DR (MHC-II; Dako North America, Inc. Carpinteria, CA) using previously described methods (Sarsoza, et al., 2009 (link)). Briefly, the protocol consisted of overnight incubation with the primary antibody, incubation with anti-mouse secondary antibody, amplification and detection with an ABC peroxidase kit and visualization with a DAB substrate kit (both from Vector Laboratories., Burlingame, CA). Sections were mounted on glass slides and coverslipped with Depex mounting media. The extent of immunolabeling was determined using “loads” as described previously (Head et al., 2008)
Wilcock D.M., Hurban J., Helman A.M., Sudduth T.L., McCarty K.L., Beckett T.L., Ferrell J.C., Murphy M.P., Abner E.L., Schmitt F.A, & Head E. (2015). Down syndrome individuals with Alzheimer's disease have a distinct neuroinflammatory phenotype compared to sporadic Alzheimer's disease. Neurobiology of aging, 36(9), 2468-2474.
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5

Immunohistochemical Analysis of Iron Deposition

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A tissue block with the most marked SWI hypointensity at the cortical GM-WM junction was resected from each of the 3 subjects with PML and 30-μm free floating sections were cut on a sliding microtome.
For total non-heme iron staining, a diaminobenzidine (DAB)-enhanced Turnbull staining protocol to detect both total ferric and ferrous non-heme iron was used.26 (link)DAB immunohistochemistry was done as previously described.27 For immunofluorescence, double labeling was done using antibodies against ferritin-light chain (ferritin-L, 10727–1-AP, rabbit polyclonal, Proteintech, Rosemont, IL, USA) with one of the following: MHCII, CD68 (Dako KP1 clone, ga60961–2, mouse monoclonal), or Iba-1 (mouse monoclonal, Cleveland Clinic Hybridoma Core); all dilutions 1:500. Secondary antibodies were Alexa Fluor donkey (anti-rabbit 488 [A21206], anti-rabbit 555 [A32794], anti-mouse 488 [A21202], and anti-mouse 555 [A31570]; Invitrogen, ThermoFisher Scientific, Waltham, MA, USA).
Mahajan K.R., Amin M., Poturalski M., Lee J., Herman D., Zheng Y., Androjna C., Howell M., Fox R.J., Trapp B.D., Jones S.E., Nakamura K, & Ontaneda D. (2021). Juxtacortical susceptibility changes in progressive multifocal leukoencephalopathy at the grey-white matter junction correlates with iron-enriched macrophages. Multiple sclerosis (Houndmills, Basingstoke, England), 27(14), 2159-2169.
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6

Immunolabeling Reactive Microglial Cells

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Endogenous peroxidase blocking (H2O2 33%, 5 minutes), followed by incubation for 30 minutes at RT with major histocompatibility complex type II (MHC II) (1/200; DAKO, Denmark) and CD68 (1/500; DAKO) primary antibodies, was used to immunolabel reactive microglial cells. EnVisionTM polymer (mouse, 30 minutes, RT; DAKO) was used as a visualization system.
Monzón M., Hernández R.S., Garcés M., Sarasa R, & Badiola J.J. (2018). Glial alterations in human prion diseases: A correlative study of astroglia, reactive microglia, protein deposition, and neuropathological lesions. Medicine, 97(15), e0320.
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