Rat anti ki67

Manufactured by BioLegend

Rat anti-Ki67 is a monoclonal antibody that binds to the Ki67 protein, a cellular marker for proliferation. It is commonly used in immunohistochemistry and flow cytometry applications to detect and quantify proliferating cells.

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4 protocols using rat anti ki67

Immunofluorescence Staining of Neural Markers

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Primary antibodies used were: mouse anti-BrdU (G3G4; DSHB), mouse anti-GFAP (Thermo Fisher Scientific), rhodamine-conjugated mouse anti-BrdU (Millipore), mouse anti-nestin (Rat-401), mouse anti-NeuN (Millipore), mouse anti–Nqo-1 (Santa Cruz Biotechnology), mouse anti-p62 (Abcam), mouse anti ubiquitin (Santa Cruz Biotechnology), mouse antivinculin (Sigma-Aldrich), rabbit anti-Atg5 (Novus Biologicals), rabbit anti-Atg16L1 (Abgent), rabbit anti-GFAP (Dako), rabbit anti-Ki67 (Spring Bioscience), rabbit anti-LC3 (Cell Signaling), rabbit anti-NBR1 (Cell Signaling), rabbit anti-Nrf2 (Abcam), rabbit anti-p62 (Enzo), rabbit anti-Sox2 (Millipore), rabbit anti-TAX1BP1 (Abgent), rabbit anti-Fip200 (ProteinTech), rabbit anti-olig2 (Millipore), rat anti-Ki67 (BioLegend), and guinea pig antidoublecortin (anti-DCX; EMD Millipore). Secondary antibodies were goat anti–rabbit IgG-FITC, goat anti–rabbit IgG–Texas red, goat anti–mouse IgG-FITC, goat anti–mouse IgG–Texas red, goat anti–mouse IgG-HRP, and goat anti–rabbit IgG-HRP (Jackson Immunology).
DHE and EUK-8 were purchased from Sigma-Aldrich. EUK-134 was purchased from Cayman Chemical. DCFDA, MitoTracker Red, and MitoTracker Green were purchased from Invitrogen.

Wang C., Chen S., Yeo S., Karsli-Uzunbas G., White E., Mizushima N., Virgin H.W, & Guan J.L. (2016). Elevated p62/SQSTM1 determines the fate of autophagy-deficient neural stem cells by increasing superoxide. The Journal of Cell Biology, 212(5), 545-560.

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Optimized Antibody Staining for Tissue Optical Clearing

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The following antibodies used in tissue optical clearing experiments were diluted in 0.5% bovine serum albumin (BSA), 10% dimethyl sulfoxide (DMSO), and 0.5% Triton-X-100 in PBS: rabbit anti-SARS-CoV N (Rockland Immunochemicals, #200–401-A50, 1:500), mouse anti-SARS-CoV NP (Sino Biological, #40143-MM05, 1:400), rat anti-I-A/I-E (Biolegend, #107601, 1:400), mouse anti-CD68 (Invitrogen, #MA5-13324, 1:400), rat anti-CD68 (BioLegend, #137001, 1:400), rat anti-Ki-67 (Biolegend, #652401, 1:200), rabbit anti-vWF (Dako, #A0082, 1:1,000), rabbit anti-Cleaved Caspase-3 (Cell Signaling, #9661, 1:200), and rabbit anti-Uteroglobin (Abcam, #ab40873, 1:500). Isotype control rabbit (Biolegend, #910801), rat (Biolegend, # 400602), and mouse (Biolegend, #401402) antibodies were used. Secondary antibodies were diluted in 2% donkey serum, 10% DMSO, and 0.5% Triton-X-100 in PBS at 1:1,000. All the following secondary antibodies were purchased from Invitrogen unless stated otherwise. Donkey anti-rabbit Alexa Fluor 568 (#A10042), donkey anti-mouse Alexa Fluor 568 (#A10037), donkey anti-rabbit Alexa Fluor 647 (#A31573), donkey anti-rat Alexa Fluor 647 (Jackson ImmunoResearch, #712–605-153), donkey anti-goat Alexa Fluor 647 (#A21447), donkey anti-rabbit Alexa Fluor 790 (#A11374), donkey anti-rat Alexa Fluor 790 (Jackson ImmunoResearch, #712–655-153).

Bagato O., Balkema-Buschmann A., Todt D., Weber S., Gömer A., Qu B., Miskey C., Ivics Z., Mettenleiter T.C., Finke S., Brown R.J., Breithaupt A, & Ushakov D.S. (2023). Spatiotemporal analysis of SARS-CoV-2 infection reveals an expansive wave of monocyte-derived macrophages associated with vascular damage and virus clearance in hamster lungs. Microbiology Spectrum, 12(1), e02469-23.

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Immunohistochemical analysis of neural markers

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Primary antibodies used were: mouse anti-GFAP (Thermo Fisher Scientific), mouse anti-nestin (Rat-401), mouse anti-p62 (Abcam), rat anti-phosphorylated p62 (MBL: D343-3), mouse anti-ubiquitin (Santa Cruz Biotechnology), rabbit anti-TBK1 (Cell Signaling), rabbit anti- phosphorylated TBK1 (Novus Biologicals), rabbit anti-GFAP (Dako), rabbit anti-Ki67 (Spring Bioscience), rabbit anti-p62 (Enzo), rabbit anti-Sox2 (Millipore), rat anti-Ki67 (BioLegend), and guinea pig anti-doublecortin (anti-DCX; EMD Millipore). Secondary antibodies were goat anti–rabbit IgG-FITC, goat anti–rabbit IgG–Texas red, goat anti–mouse IgG-FITC, goat anti–mouse IgG–Texas red, goat anti–mouse IgG-HRP, and goat anti–rabbit IgG-HRP (Jackson Immunology). Dihydroethidium (DHE) was purchased from Sigma-Aldrich and Amlexanox was purchased from MedChemExpress. Transfections were carried out using Lipofectamine 3000 reagent (Invitrogen).

Liu H., Wang C., Yi F., Yeo S., Haas M., Tang X, & Guan J.L. (2021). Non-canonical function of FIP200 is required for neural stem cell maintenance and differentiation by limiting TBK1 activation and p62 aggregate formation. Scientific Reports, 11, 23907.

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Comprehensive Immunohistochemical Analysis of Intestinal Tissues

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Ileum tissues were fixed with 4% PFA and dehydrated with 15%, and then 30% sucrose in PBS. Dehydrated tissues were formed into a Swiss roll, frozen, and sliced. For staining, collected organoids were permeabilized in PBS containing 0.1% Tween 20 and blocked with 0.5% BSA in PBS for 1 h. For Muc2 staining, ileum tissues containing feces were fixed in Carnoy’s solution and embedded in paraffin. The primary antibodies used were rabbit anti-Muc2 (Abcam), rat anti-Ki67 (Biolegend), rabbit anti-lysozyme (Abcam), goat anti-Wnt3 (Abcam), mouse anti-β-catenin (BD Bioscience), and mouse anti-phospho-ERK1/2 (Thermo Fisher Scientific). Secondary antibodies were Alexa Fluor goat 594 anti-rat IgG (Biolegend), Alexa Fluor 488 goat anti-mouse IgG (Abcam), Alexa Fluor 546 donkey anti-goat IgG (Thermo Fisher Scientific), and Alexa Fluor 488 goat anti-rabbit IgG (Thermo Fisher Scientific).

Kim S., Shin Y.C., Kim T.Y., Kim Y., Lee Y.S., Lee S.H., Kim M.N., O E., Kim K.S, & Kweon M.N. (2021). Mucin degrader Akkermansia muciniphila accelerates intestinal stem cell-mediated epithelial development. Gut Microbes, 13(1), 1892441.

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