Ki67 alexa fluor 647

Manufactured by BD

Ki67 Alexa Fluor 647 is a fluorescently-labeled antibody used for the detection and quantification of the Ki67 protein, a cellular marker of proliferation. The Alexa Fluor 647 dye is used to label the antibody, enabling its visualization through fluorescence microscopy or flow cytometry techniques.

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Lab products found in correlation

Cd34 fitc, by BD (1 mentions) Cytofix buffer, by BD (1 mentions) Lsr 2, by BD (1 mentions) Foxp3 staining buffer set, by Miltenyi Biotec (1 mentions) Cd45 percp cy5.5 clone 30 f11, by Thermo Fisher Scientific (1 mentions) Cd31 fitc clone 390, by Thermo Fisher Scientific (1 mentions) Gp38 pe clone 8, by Thermo Fisher Scientific (1 mentions) Epcam pe cy7 clone g8, by BioLegend (1 mentions) Cyan adp, by Agilent Technologies (1 mentions)

Spelling variants of this product

Alexa Fluor 647 (58 citations) Alexa 647 (7 citations) Alexa Fluor 647 Mouse anti-Ki67 (2 citations)

4 protocols using ki67 alexa fluor 647

Multiparametric Flow Cytometry Protocol

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Flow cytometry for surface marker analysis was performed using similar procedures as cell sorting (see “Cell preparation”) and analyzed on a BD Cytoflex or BD LSRII. Fluorochromeconjugated antibodies are listed in the Key Resources Table. Cell cycle analysis was performed as follows: (1) stain MUTZ-3 cells with CD14-PE-Cy7 (Coulter A22331) and CD34-FITC (BD Pharmingen 348053) antibodies; (2) spin and permeabilize using Phosphoflow Perm 2 solution (BD 347692 diluted 10× in H₂O); (3) wash with PBS 2% FBS; (4) stain with Ki67 Alexa Fluor 647 (BD Pharmingen 561126); (4) wash; (5) resuspend in Cytofix buffer (BD 554655 diluted 4× in PBS) with DAPI at 1 μg / ml; (5) analysis on an BD LSRII analyzer within 30 minutes. Data was analyzed using FlowJo software (Tree Star, Inc.).

van Galen P., Hovestadt V., Wadsworth M I.I., Hughes T., Griffin G.K., Battaglia S., Verga J.A., Stephansky J., Pastika T.J., Lombardi Story J., Pinkus G.S., Pozdnyakova O., Galinsky I., Stone R.M., Graubert T.A., Shalek A.K., Aster J.C., Lane A.A, & Bernstein B.E. (2019). Single-cell RNA-seq reveals AML hierarchies relevant to disease progression and immunity. Cell, 176(6), 1265-1281.e24.

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PD-1 Blockade Enhances PBMC Proliferation

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Stimulated PBMC were cultured in the presence of PD-1 blocking or isotype control antibodies and rhIL-2 for 72 h as previously described (24 (link)). The proliferation was accessed by intracellular staining for the cell cycle related nuclear antigen Ki67 Alexa Fluor 647 (BD Bioscience) that was performed with Foxp3 Staining Buffer Set (Miltenyi Biotec) and analyzed by flow cytometric analysis.

Garcia de Moura R., Covre L.P., Fantecelle C.H., Gajardo V.A., Cunha C.B., Stringari L.L., Belew A.T., Daniel C.B., Zeidler S.V., Tadokoro C.E., de Matos Guedes H.L., Zanotti R.L., Mosser D., Falqueto A., Akbar A.N, & Gomes D.C. (2021). PD-1 Blockade Modulates Functional Activities of Exhausted-Like T Cell in Patients With Cutaneous Leishmaniasis. Frontiers in Immunology, 12, 632667.

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Multiparametric Flow Cytometry Analysis

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Single-cell suspensions were stained for 30 min in PBS (with 0.5% BSA and 2 mM EDTA) with cocktails of the following Abs: CD31-FITC clone 390, gp38-PE clone 8.1.1, CD45 PerCP/Cy5.5 clone 30-F11 (from eBioscience), epithelial cell adhesion molecule (EPCAM) PE/Cy7 clone G8.8 (from BioLegend), Ki67–Alexa Fluor 647, and BrDU–Alexa Fluor 647 (BD Pharmingen). Afterwards cells were washed twice, resuspended, and then analyzed using a CyAn ADP (Dako) with forward/side scatter gates set to exclude nonviable cells. Data were analyzed with FlowJo software (Tree Star).

Nayar S., Campos J., Chung M.M., Navarro-Núñez L., Chachlani M., Steinthal N., Gardner D.H., Rankin P., Cloake T., Caamaño J.H., McGettrick H.M., Watson S.P., Luther S., Buckley C.D, & Barone F. (2016). Bimodal Expansion of the Lymphatic Vessels Is Regulated by the Sequential Expression of IL-7 and Lymphotoxin α1β2 in Newly Formed Tertiary Lymphoid Structures. The Journal of Immunology Author Choice, 197(5), 1957-1967.

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Proliferation of Activated NK Cells

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Sorted NK cells were cultured in the presence of rhIL-2 (100 U/ml) for 48 hr, followed by intracellular staining for ª 2020 John Wiley & Sons Ltd, Immunology, 159, 429-440 the cell-cycle-related nuclear antigen Ki67 Alexa Fluor 647 (B56; BD Bioscience) and analysed by flow cytometry. Supplemented culture medium without rIL-2 was used as control.

Covre L.P., Devine O.P., Garcia de Moura R., Vukmanovic-Stejic M., Dietze R., Ribeiro-Rodrigues R., Guedes H.L.M., Lubiana Zanotti R., Falqueto A., Akbar A.N, & Gomes D.C.O. (2020). Compartmentalized cytotoxic immune response leads to distinct pathogenic roles of natural killer and senescent CD8⁺ T cells in human cutaneous leishmaniasis. Immunology, 159(4).

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