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Anti human cd45 clone hi30

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Anti-human CD45 (clone HI30) is a laboratory reagent used to detect and quantify human CD45-positive cells. CD45 is a pan-leukocyte marker expressed on the surface of most human hematopoietic cells. This monoclonal antibody can be used in various applications, such as flow cytometry, to identify and characterize different types of white blood cells.

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Lab products found in correlation

Anti mouse cd45 clone 30 f11, by BD (3 mentions) Anti ki67 clone 8d5, by Cell Signaling Technology (2 mentions) Alexa fluor 568 goat anti rabbit igg h l, by Thermo Fisher Scientific (2 mentions) Alexa fluor 488 goat anti mouse igg h l, by Thermo Fisher Scientific (2 mentions) Anti human mitochondria clone 113 1, by Merck Group (2 mentions) Annexin 5 fitc and, by BD (1 mentions) Anti human cd3, by BioLegend (1 mentions) Fc500, by Beckman Coulter (1 mentions) Rbc lysis buffer, by BioLegend (1 mentions) Anti human cd8 clone rpa t8, by BD (1 mentions) Anti human cd38 clone hit2, by BioLegend (1 mentions) Cd45ra clone hi100, by Thermo Fisher Scientific (1 mentions) Antihuman cd3 clone ucht1, by BD (1 mentions) Affinipure f ab 2 fragment goat anti mouse igg, by Jackson ImmunoResearch (1 mentions) Lsrfortessa, by BD (1 mentions)

5 protocols using anti human cd45 clone hi30

1

Immunofluorescence Staining of Cells

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Cells were fixed, permeabilized, blocked against nonspecific adhesion, immunostained for target proteins, and then imaged on an inverted Eclipse Ti epifluorescence microscope (Nikon). Primary antibodies were administered at manufacturer recommended concentration: anti-human CD45 (clone HI30, Becton Dickinson), anti-mouse CD45 (clone 30-F11, Becton Dickinson), anti-human mitochondria (clone 113–1, MilliporeSigma), anti-Ki67 (clone 8D5, Cell Signaling Technology), and anti-phospho-histone H2A.X (Ser139, clone 20E3, Cell Signaling Technology). The secondary antibodies used were Alexa Fluor 488 goat anti-mouse IgG (H+L) (Invitrogen) and Alexa Fluor 568 goat anti-rabbit IgG (H+L) (Invitrogen).
Yankaskas C.L., Thompson K.N., Paul C.D., Vitolo M.I., Mistriotis P., Mahendra A., Bajpai V.K., Shea D.J., Manto K.M., Chai A.C., Varadarajan N., Kontrogianni-Konstantopoulos A., Martin S.S, & Konstantopoulos K. (2019). A microfluidic assay for the quantification of the metastatic propensity of breast-cancer specimens. Nature biomedical engineering, 3(6), 452-465.
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2

Immunofluorescence Staining of Cells

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Cells were fixed, permeabilized, blocked against nonspecific adhesion, immunostained for target proteins, and then imaged on an inverted Eclipse Ti epifluorescence microscope (Nikon). Primary antibodies were administered at manufacturer recommended concentration: anti-human CD45 (clone HI30, Becton Dickinson), anti-mouse CD45 (clone 30-F11, Becton Dickinson), anti-human mitochondria (clone 113–1, MilliporeSigma), anti-Ki67 (clone 8D5, Cell Signaling Technology), and anti-phospho-histone H2A.X (Ser139, clone 20E3, Cell Signaling Technology). The secondary antibodies used were Alexa Fluor 488 goat anti-mouse IgG (H+L) (Invitrogen) and Alexa Fluor 568 goat anti-rabbit IgG (H+L) (Invitrogen).
Yankaskas C.L., Thompson K.N., Paul C.D., Vitolo M.I., Mistriotis P., Mahendra A., Bajpai V.K., Shea D.J., Manto K.M., Chai A.C., Varadarajan N., Kontrogianni-Konstantopoulos A., Martin S.S, & Konstantopoulos K. (2019). A microfluidic assay for the quantification of the metastatic propensity of breast-cancer specimens. Nature biomedical engineering, 3(6), 452-465.
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3

Peripheral Blood Lymphocyte Engraftment Assessment

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Peripheral blood was collected via saphenous vein using heparin-coated capillary tubes, and was examined for engraftment of human lymphocytes. Erythrocytes were removed by RBC Lysis Buffer (BioLegend, San Diego, CA). Cells were washed with PBS (containing 2% fetal bovine serum [FBS] and 2.5 mM EDTA) and stained for 30 minutes at 4°C using the following mouse monoclonal antibodies: anti-human CD3 (clone HIT3a), CD19 (clone HIB19), CD41 (clone HIP8), anti-mouse CD41 (clone MWReg30) (BioLegend, San Diego, CA), anti-human CD45 (clone HI30) and anti-mouse CD45.1 (clone A20, BD Biosciences, San Jose, CA). Data was acquired on an FC500 (Beckman Coulter) or LSRII (BD Biosciences, San Jose, CA) and was analyzed using FlowJo software (Treestar, Ashland, OR, USA). Donor cells were stained for zombie aqua, anti-human CD3 (BioLegend, San Diego, CA), Annexin V FITC and mouse monoclonal anti-human CD34 (clone 581, BD Biosciences, San Jose, CA) to assess the viability according to the manufacturer’s protocol.
Fujii H., Luo Z.J., Kim H.J., Newbigging S., Gassas A., Keating A, & Egeler R.M. (2015). Humanized Chronic Graft-versus-Host Disease in NOD-SCID il2rγ-/- (NSG) Mice with G-CSF-Mobilized Peripheral Blood Mononuclear Cells following Cyclophosphamide and Total Body Irradiation. PLoS ONE, 10(7), e0133216.
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4

Adoptive Transfer of CD8+ TILs for Melanoma

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NOD-scid IL2Rgammanull (NSG) mice were engrafted with either 5 × 106 MEL 526 MEL tumor cells or 5 × 106 autologous primary melanoma tumor cells. On day 12, either 10 ×106 sorted CD8+BTLA+ or sorted CD8+BTLA− were adoptively transferred into tumor-bearing mice (n= 5 to 8 per group). Recombinant human IL-2 (Proleukin, Prometheus) was administered intraperitoneally at a concentration of 6 ×105 I.U. immediately after TIL transfer and daily for three days. Tumor size was measured every other day. Mice were sacrificed when tumors exceeded 15 mm diameter. Peripheral blood was collected every other day, lysed with ACK lysis buffer, then stained for AQUA (Invitrogen), anti-human CD45 (clone HI30, BD Pharmingen™), and anti-human CD8 (clone RPA-T8, BD Pharmingen™).
Ritthipichai K., Haymaker C.L., Martinez M., Aschenbrenner A., Yi X., Zhang M., Kale C., Vence L.M., Roszik J., Hailemichael Y., Overwijk W.W., Varadarajan N., Nurieva R., Radvanyi L.G., Hwu P, & Bernatchez C. (2017). Multifaceted role of BTLA in the control of CD8+ T cell fate after antigen encounter. Clinical cancer research : an official journal of the American Association for Cancer Research, 23(20), 6151-6164.
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5

Flow Cytometric Characterization of CAR T-Cells

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Flow cytometry was performed on BD LSR Fortessa. Transduction efficiency was measured with an APC-conjugated antibody toward NGFR (CD271; clone ME20.4, BioLegend) and Protein L for CCR, whereas for CARs it was measured in the phycoerythrin (PE) channel to detect dsRed and AffiniPure F(ab’)2 Fragment Goat Anti-Mouse IgG (Jackson ImmunoResearch). The following antibodies were used for flow cytometry staining: Anti-Human CD3 (clone UCHT1, BD Biosciences), Anti-Human CD4 (clone L200, BD Biosciences), Anti-Mouse CD45 (clone 30-F11, BD Biosciences), Anti-Human CD8 (clone SK1, BD Biosciences), Anti-Human CD45 (clone HI30, BD Biosciences), Anti-human CD366 (TIM-3) (clone F38-2E2, BioLegend or clone F38-2E2, Thermo Fisher Scientific), Anti-human CD279 (PD-1) (clone EH12.2H7, BioLegend or clone eBioJ105, Thermo Fisher Scientific), Anti-human CD62L (clone DREG-56, BioLegend), Anti-human CD269 (BCMA) (clone 19F2, BioLegend), Anti-human CD38 (clone HIT2, BioLegend), CD45RA (clone HI100, Thermo Fisher Scientific), LAG-3 (CD223; clone 3DS223H, Thermo Fisher Scientific) (staining 1 μg per mL of sample, incubation for 30 minutes at 4°C in PBS with 1% BSA, 10% FBS and 0.1% NaN3 sodium azide). For the quantification of the CD38, the Quantibrite Phycoerythrin (PE) Fluorescence Quantitation Kit (BD) was used. Flow cytometry data analysis was performed with FCS Express 6 flow software.
Katsarou A., Sjöstrand M., Naik J., Mansilla-Soto J., Kefala D., Kladis G., Nianias A., Ruiter R., Poels R., Sarkar I., Patankar Y.R., Merino E., Reijmers R.M., Frerichs K.A., Yuan H., de Bruijn J., Stroopinsky D., Avigan D., van de Donk N.W., Zweegman S., Mutis T., Sadelain M., Groen R.W, & Themeli M. (2021). Combining a CAR and a chimeric costimulatory receptor enhances T cell sensitivity to low antigen density and promotes persistence. Science translational medicine, 13(623), eabh1962.
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