Ki67 pe clone b56

Manufactured by BD

Sourced in United States

Ki67-PE (clone B56) is a fluorescently-labeled antibody that binds to the Ki-67 protein, a marker of cellular proliferation. This product is intended for research use only.

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Lab products found in correlation

Foxp3 staining kit, by Thermo Fisher Scientific (2 mentions) Anti cd4 percp cy5.5 clone rm4 5, by BD (1 mentions) Anti cd44 apc cy7 clone im7, by BD (1 mentions) Fortessa flow cytometer, by Tree Star (1 mentions) 40 μm gauze, by BD (1 mentions) Foxp3 alexafluor clone fjk 16a, by Thermo Fisher Scientific (1 mentions) Dispase 1, by Roche (1 mentions) Dnase 1, by Merck Group (1 mentions) Collagenase d, by Roche (1 mentions) Streptavidin apc, by BD (1 mentions) Cd158b ch l, by BD (1 mentions) Lsrii flow cytometer, by BD (1 mentions) Staphylococcal enterotoxin b, by Toxin Technology (1 mentions) Cd3 apc cy7 clone sk7, by BioLegend (1 mentions) Facsaria flow cytometer, by BD (1 mentions)

4 protocols using ki67 pe clone b56

Multi-parameter flow cytometry analysis of T cells

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Blood was collected by retro-orbital puncture. Spleen cells were obtained by mechanical disruption. After Fc receptor blocking (clone 2.4G2, 1:1,000; BD Biosciences), cells were stained with anti-CD4 PerCP-Cy5.5 (clone RM4-5; 1:800; BD Biosciences) or CD4 BV785 (clone RM4-5; 1:800; BioLegend), anti-TCR-β APC (clone H57-597; 1:200; BD Biosciences) or PE (clone H57-597; 1:100; BD Biosciences), anti-NKp46 FITC (clone 29A1.4; 1:300; eBiosciences), anti-CD8 Alexa700 (clone 53-6.7; 1:1,600; BioLegend), anti-CD25 PE-Cy7 (clone PC61; 1:300; eBiosciences), anti-CD62L BV605 (clone MEL-14; 1:800; BioLegend) and anti-CD44 APC-Cy7 (clone IM7; 1:400; BD Biosciences) in PBS/3% FCS. Then anti-Foxp3 eFluor450 (clone FJK-16 s; 1:150; eBioscience) and Ki67-PE (clone B56; 1:50; BD Biosciences) or Ki67-Alexa647 (clone B56; 1:50; BD Biosciences) staining was performed using the eBioscience Foxp3 staining kit according to the manufacturer's instructions. Cells were acquired on a Fortessa flow cytometer and analysed with FlowJo software (TreeStar).

Pérol L., Lindner J.M., Caudana P., Nunez N.G., Baeyens A., Valle A., Sedlik C., Loirat D., Boyer O., Créange A., Cohen J.L., Rogner U.C., Yamanouchi J., Marchant M., Leber X.C., Scharenberg M., Gagnerault M.C., Mallone R., Battaglia M., Santamaria P., Hartemann A., Traggiai E, & Piaggio E. (2016). Loss of immune tolerance to IL-2 in type 1 diabetes. Nature Communications, 7, 13027.

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Isolation and Characterization of Intestinal T Cells

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Lungs were digested as above. Colons were incubated with PBS containing EDTA at 37°C for 15 min then digested twice for 35 min with a cocktail of collagenase D (1 mg/ml, Roche), dispase I (0.1 mg/ml, Roche), and DNase I (333 μg/ml, Sigma), smashed and filtered through a 40 μm gauze (BD Biosciences). Single-cell suspensions were then stained for flow cytometry with the following antibodies: CD4 Pacific blue (clone GK1.5), CD103 biotin (clone M290 or 2E7), CD25 PE-cy7 (clone PC61), PD1 PE (clone RMP1-30), GITR FITC (clone DTA-1), and streptavidin APC were purchased from BD Biosciences or Biolegend. FoxP3 AlexaFluor (clone FJK-16a, eBioscience) and Ki-67 PE (clone B56, BD Biosciences) were used in combination with a FoxP3 staining kit (eBioscience).

Zaiss M.M., Rapin A., Lebon L., Dubey L.K., Mosconi I., Sarter K., Piersigilli A., Menin L., Walker A.W., Rougemont J., Paerewijck O., Geldhof P., McCoy K.D., Macpherson A.J., Croese J., Giacomin P.R., Loukas A., Junt T., Marsland B.J, & Harris N.L. (2015). The Intestinal Microbiota Contributes to the Ability of Helminths to Modulate Allergic Inflammation. Immunity, 43(5), 998-1010.

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Multiparameter Immune Cell Profiling

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All rested samples were stained with viability dye and surface antibodies for 30 minutes at 4°C. Then cells were washed and permeabolized using the FOXp3 transcription Factor Staining Set for staining with Ki67 (00-5523, eBioscience, San Diego, CA). The following mouse anti-human antibodies were used: CD3-PE-eFlour 610 (clone UCHT1), CD19-PE-eFlour 610 (clone 61D3), CD14-PE-eFlour 610 (clone HIB19), and CD57-eFlour 450 (clone TBO1) from eBioscience; CD56-BV605 (clone NCAM) and CD117-BV650 (clone 104D2) from Biolegend (San Diego, CA); NKG2A-APC (clone Z199) from Beckman Coulter (Indianapolis, IN); Near-IR LIVE/DEAD Fixable dead cell stain from Life Technologies (San Deigo, CA); and Ki67-PE (clone B56) as part of Transcription Factor Binding set and cocktail of FITC-conjugated KIR antibodies against NKB1 (clone DX9), CD158a (clone HP-3E4), and CD158b (CH-L) from BD Biosciences (San Jose, CA). All flow cytometry was collected on a LSR II flow cytometer (BD Biosciences) and analyzed using Flowjo v10 (Flowjo LLC, Ashland, OR).

Bergerson R.J., Williams R., Wang H., Shanley R., Colbenson G., Kerber A., Cooley S., Curtsinger J.M., Felices M., Miller J.S, & Verneris M.R. (2016). Fewer circulating natural killer cells 28 days after double cord blood transplantation predicts inferior survival and IL-15 response. Blood Advances, 1(3), 208-218.

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T-cell Proliferation and Chemokine Expression

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MSCs were seeded onto 96-well plates with appropriate densities. PBMCs from random donor were added into each well with the final concentration of 0.1 × 10⁶ cells per well. 500 ng/mL staphylococcal enterotoxin B (SEB) (Toxin Technology, Sarasota, FL) was used to stimulate the T cells in the PBMCs. Ki67 proliferation assay was performed to measure T-cell proliferation after 4 days. For Ki67 proliferation assay, intracellular flow cytometry staining was performed with BD Cytofix and Cytoperm procedure with the antibodies CD3APCCy7 (Clone SK7) (Biolegend, USA) and Ki67 PE (Clone B56) (BD Biosciences, USA) according to manufacturer instructions (BD Biosciences, San Jose, CA). Samples were acquired in BD FACS Aria flow cytometer, and the results were analyzed in FlowJo software. For non-contact cocultures, MSCs are cultured on the bottom of the well and SEB-activated PBMCs were cultured on the transwell membrane (0.4 µM). Three days later MSCs were lysed and RNA was isolated. Total RNA is converted into cDNA (Qiagen, USA). Quantitative real-time PCR was performed to identify the expression of chemokines.

Lipat A.J., Cottle C., Pirlot B.M., Mitchell J., Pando B., Helmly B., Kosko J., Rajan D., Hematti P, & Chinnadurai R. (2022). Chemokine Assay Matrix Defines the Potency of Human Bone Marrow Mesenchymal Stromal Cells. Stem Cells Translational Medicine, 11(9), 971-986.

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