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Mouse cd45 clone 30 f11

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Mouse CD45 (clone 30-F11) is a cell surface marker commonly used in flow cytometry applications to identify and characterize mouse leukocytes. It is a pan-leukocyte antigen expressed on all cells of hematopoietic origin, including T cells, B cells, monocytes, and granulocytes.

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Lab products found in correlation

Lsrfortessa, by BD (4 mentions) Human cd45 clone hi30, by BioLegend (3 mentions) Zombie yellow fixable viability kit, by BioLegend (3 mentions) Cd3 clone okt3, by BioLegend (2 mentions) Cd8 clone sk1, by BioLegend (2 mentions) Cd45ro clone uchl1, by BioLegend (2 mentions) Cd45ra hi100, by BioLegend (2 mentions) Cd4 clone a161a1, by BioLegend (2 mentions) Pd 1 clone mih4, by BD (2 mentions) Countbright absolute counting beads, by Thermo Fisher Scientific (1 mentions) Anti bcl 2 clone 100, by BioLegend (1 mentions) Lsr 2 fortessa cytometer, by BD (1 mentions) Human cd3 clone okt3, by BioLegend (1 mentions) Cd27 clone m t271, by BioLegend (1 mentions) Tim 3 clone f38 2e2, by BioLegend (1 mentions) Cd62l dreg 56, by BioLegend (1 mentions) Cd19 clone hib19, by BioLegend (1 mentions) Foxp3 fixation permeabilization kit, by Thermo Fisher Scientific (1 mentions) Rbc lysis buffer, by BioLegend (1 mentions) Live dead fixable aqua dead cell stain kit, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

CD45 (30-F11, (72 citations) CD45 (clone 30-F11, (62 citations) Clone 30-F11 (48 citations) Anti-mouse CD45 (clone 30-F11, (9 citations) PE-conjugated anti-mouse CD45 antibody (clone 30-F11, (2 citations) Anti-mouse CD45 antibody clone: 30-F11 (2 citations) Anti-mouse CD45-PerCP-Cy5.5 (clone 30-F11; (2 citations) Anti-mouse CD45-PE monoclonal antibody (clone 30-F11, (2 citations)

5 protocols using mouse cd45 clone 30 f11

1

T Cell Phenotyping Protocol

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All samples were analyzed with an LSR Fortessa (BD Bioscience), and data were analyzed using FlowJo software (FlowJo). T cell phenotype was evaluated via Zombie Yellow Fixable Viability Kit (BioLegend), human CD45 (clone HI30; BioLegend), mouse CD45 (clone 30F11; BioLegend), CD3 (clone OKT3; BioLegend), CD4 (clone A161A1; BioLegend), CD8 (clone SK1; BioLegend), PD-1 (clone MIH4; BD Bioscience), CD45RO (clone UCHL1; BioLegend), and CD45RA (HI100; BioLegend).
Wang Y., Buck A., Grimaud M., Culhane A.C., Kodangattil S., Razimbaud C., Bonal D.M., Nguyen Q.D., Zhu Z., Wei K., O'Donnell M.L., Huang Y., Signoretti S., Choueiri T.K., Freeman G.J., Zhu Q, & Marasco W.A. (2021). Anti-CAIX BBζ CAR4/8 T cells exhibit superior efficacy in a ccRCC mouse model. Molecular Therapy Oncolytics, 24, 385-399.
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2

Multiparametric Flow Cytometry for Cell Phenotyping

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Cells were resuspended in FACS staining buffer (PBS + 2% fetal bovine serum) using the following antibodies: human CD3 (clone OKT3, Biolegend), anti-BCL-2 (clone 100, Biolegend), human CD45 (clone 2D1, Biolegend), mouse CD45 (clone 30-F11, Biolegend). CART19 was detected using PE-conjugated anti-CAR19 idiotype antibody (Novartis). To monitor caspase 3/7 activity, CellEvent™ Caspse3/7 Green Read Flow™ reagent was used by following a manufacturing protocol. To determine the absolute cell numbers (tumor or T cells) acquired during flow cytometry, CountBright absolute counting beads (ThermoFisher) were used. Cell viability was established using Live/Dead Aqua or violet fixable staining kit (ThermoFisher), Propidium iodide (PI) and 7-Aminoacctinomycine D (7-AAD), and data were acquired on an LSRII Fortessa Cytometer (BD). Intracellular staining was performed by using fixation/pemeabilization buffer and following manufacturing protocol. All data analysis was performed using FlowJo 9.0 or 10 software (FlowJo, L.L.C., BD, Ashland, OR).
Lee Y.G., Guruprasad P., Ghilardi G., Pajarillo R., Sauter C.T., Patel R., Ballard H.J., Hong S.J., Chun I., Yang N., Amelsberg K.V., Cummins K.D., Svoboda J., Gill S., Chong E.A., North K., Church S.E., Fraietta J.A., Chang W.J., Lacey S.F., Lu X.M., Zhang Y., Whig K., Schultz D.C., Cherry S., Gerson J., Schuster S.J., Porazzi P, & Ruella M. (2022). Modulation of BCL-2 in both T Cells and Tumor Cells to Enhance Chimeric Antigen Receptor T cell Immunotherapy against Cancer. Cancer discovery, 12(10), 2372-2391.
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3

Evaluating CAR-T Cell Viability

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CAR-T expansion was evaluated using Zombie Yellow™ Fixable Viability Kit (BioLegend), human CD45 (clone HI30, BioLegend), and mouse CD45 (clone 30F11, BioLegend). All samples were analyzed with an LSR Fortessa (BD Bioscience) and data was analyzed using FlowJo software (FlowJo LLC).
Wang Y., Buck A., Piel B., Zerefa L., Murugan N., Coherd C.D., Miklosi A.G., Johal H., Bastos R.N., Huang K., Ficial M., Laimon Y.N., Signoretti S., Zhong Z., Hoang S.M., Kastrunes G.M., Grimaud M., Fayed A., Yuan H.C., Nguyen Q.D., Thai T., Ivanova E.V., Paweletz C.P., Wu M.R., Choueiri T.K., Wee J.O., Freeman G.J., Barbie D.A, & Marasco W.A. (2024). Affinity fine-tuning anti-CAIX CAR-T cells mitigate on-target off-tumor side effects. Molecular Cancer, 23, 56.
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4

Comprehensive T cell Phenotyping Protocol

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All samples were analyzed with an LSR Fortessa (BD Bioscience) and data were analyzed using FlowJo software (FlowJo LLC). T cell phenotype was evaluated via:Zombie Yellow Fixable Viability Kit (BioLegend), human CD45 (clone HI30, BioLegend), mouse CD45 (clone 30F11, BioLegend), CD3 (clone OKT3, BioLegend), CD4 (clone A161A1, BioLegend), CD8 (clone SK1, BioLegend), CD19 (clone HIB19, BioLegend), CD27 (clone M-T271, BioLegend), CD62L (DREG-56, BioLegend), PD-1 (clone MIH4, BD Bioscience), TIM-3 (clone F38-2E2, BioLegend), CD45RO (clone UCHL1, BioLegend), CD45RA (HI100, BioLegend).
Wang Y., Cho J.W., Kastrunes G., Buck A., Razimbaud C., Culhane A.C., Sun J., Braun D.A., Choueiri T.K., Wu C.J., Jones K., Nguyen Q.D., Zhu Z., Wei K., Zhu Q., Signoretti S., Freeman G.J., Hemberg M, & Marasco W.A. (2024). Immune-restoring CAR-T cells display antitumor activity and reverse immunosuppressive TME in a humanized ccRCC mouse model. iScience, 27(2), 108879.
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5

Isolation and Characterization of Pancreatic Tumor Immune Cells

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Spleen and pancreatic tumors were harvested from tumor bearing mice, minced in 3ml lysis buffer (HBSS+ 50U/ml DnaseI + 0.5mg/ml Collagenase D + 2.5mM MgCl2) and incubated at 37 °C for 30 minutes. Tissue chunks were ground with rubber grinder and passed through a 70 μm cell restrainer. Cells were incubated with 5ml 1x RBC lysis buffer (420301, Biolegend, CA) for 4 minutes, then diluted in 2%FBS in 1xPBS to a final volume of 30ml and spun down. Live cells were determined by LIVE/DEAD® fixable aqua dead cell stain kit (Molecular Probes). The cell pellets were re-suspended in PBS with 2% FBS for Fluorescence-activated cell sorting (FACS) analysis. Cells were stained with cell surface markers as indicated followed by fixation/permeabilization using foxp3 fixation/permeabilization kit (eBioscience). Cells were imaged on BD LSRFortessa (BD Biosciences) and analyzed using FlowJo software (Tree Star). Pancreas infiltrating immune cells were stained with different combinations of fluorochrome-coupled antibodies against mouse CD45 (clone 30-F11, Biolegend), F4/80 (clone BM8, Biolegend).
Yang A., Herter-Sprie G., Zhang H., Lin E.Y., Biancur D., Wang X., Deng J., Hai J., Yang S., Wong K.K, & Kimmelman A.C. (2018). Autophagy sustains pancreatic cancer growth through both cell autonomous and non-autonomous mechanisms. Cancer discovery, 8(3), 276-287.
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