Anti cd45

Manufactured by Beckman Coulter

Sourced in United States

Anti-CD45 is a laboratory reagent used to detect and identify cells expressing the CD45 antigen, which is present on the surface of most hematopoietic cells. It is a widely used marker for the identification and enumeration of leukocytes in various clinical and research applications.

Automatically generated - may contain errors

Lab products found in correlation

Spelling variants of this product

Anti-CD45-APC (6 citations) Anti-CD45-Krome orange (6 citations) FITC anti-human CD45 (5 citations) Anti-CD45-PC7 (5 citations) Anti-CD45-PeCy7 (3 citations) Anti-CD45-FITC antibodies (2 citations) Anti-CD45 PE (2 citations) Anti-human CD45 monoclonal antibody (2 citations) FITC conjugated anti-CD45 (2 citations) Anti-human-CD45-KO (J33, (2 citations)

9 protocols using anti cd45

Identification and Characterization of MDSC Subsets

Check if the same lab product or an alternative is used in the 5 most similar protocols

The MDSC subsets, G-MDSCs (CD15⁺CD33⁺CD11b⁺CD14⁻HLA⁻DR^−/low) and M-MDSCs (CD14⁺CD15⁻CD11b⁺CD33⁺HLA⁻DR^−/low), were characterized by FACS using a CytoFLEX (Beckman Coulter) with a panel of the following fluorescent-labeled monoclonal antibodies: anti-CD15, anti-CD33, anti-CD14, anti-CD45, anti-CD11b, and anti-HLA-DR. All antibodies were purchased from eBioscience (Invitrogen). The frequencies of cells expressing PD-L1 and PD-L2 within the MDSC subsets were determined using fluorescent-labeled monoclonal antibodies CD274-APC and CD273-PE. This allowed for the identification of PD-L1⁺ G-MDSCs, PD-L1⁺ M-MDSCs, PD-L2⁺ G-MDSCs, and PD-L2⁺ M-MDSCs. The flow cytometric analyses strategies for identifying G-MDSCs and M-MDSCs in the peripheral blood are presented in Figures 2 and 3.
Figure 2

The strategies of flow cytometric analyses to identify G-MDSCs in the circulation. G-MDSCs, granulocytic myeloid-derived suppressor cells.

Figure 3

The strategies of flow cytometric analyses to identify M-MDSCs in the circulation. M-MDSCs, monocytic myeloid-derived suppressor cells.

Liu Z., Zhang M., Shi X., Zhao W., Cao C., Jin L., Wang Y, & Xiao J. (2023). Decreased programmed cell death ligand 2-positive monocytic myeloid-derived suppressor cells and programmed cell death protein 1-positive T-regulatory cells in patients with type 2 diabetes: implications for immunopathogenesis. Endocrine Connections, 12(9), e230218.

+ Open protocol

+ Expand

Multiparametric Flow Cytometry Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Surface phenotypes were determined using an Epics XL MCL (Beckman Coulter, Brea, CA, USA). The following monoclonal antibodies (mAbs) were purchased from Beckman Coulter: anti‐CD3, anti‐CD4, anti‐CD8, anti‐Vγ9TCR, anti‐CD14, anti‐CD25, anti‐CD45, anti‐CD54, anti‐CD56, anti‐HLA‐DR, anti‐CD40, anti‐CD80, anti‐CD86, anti‐CD11c, anti‐CD36, mouse immunoglobulin (Ig)G1, mouse IgG2 and mouse IgG2b mAbs. Anti‐HLA‐class 1 and anti‐CCR7 mAbs were purchased from Beckton Dickinson (San Jose, CA, USA) and R&D Systems (Minneapolis, MN, USA), respectively. Anti‐TCR Vα24TCR and anti‐TCR Vβ11 mAbs were purchased from Beckman Coulter (Villepinte, France). Anti‐human CD273 (PD‐L2) and CD274 (PD‐L1) mAbs were purchased form eBioscience (San Diego, CA, USA). Anti‐human CD152 (CTLA‐4) and CD279 (PD‐1) mAbs were purchased from BioLegend (San Diego, CA, USA). Anti‐FoxP3 mAb for intracellular staining was purchased from BD Biosciences (Tokyo, Japan). All mAbs were conjugated with fluorescein isothiocyanate (FITC), phycoerythrin (PE), antigen‐presenting cells (APC), extracellular domain (ECD), proprotein convertase (PC)5 or PC7. Z was purchased from Novartis Pharmaceuticals (Basel, Switzerland) and G from Funakoshi Co. Ltd (Tokyo, Japan).

Eiraku Y., Terunuma H., Yagi M., Deng X., Nicol A.J, & Nieda M. (2018). Dendritic cells cross‐talk with tumour antigen‐specific CD8+ T cells, Vγ9γδT cells and Vα24NKT cells in patients with glioblastoma multiforme and in healthy donors. Clinical and Experimental Immunology, 194(1), 54-66.

+ Open protocol

+ Expand

Monocyte Immunophenotyping by Flow Cytometry

Check if the same lab product or an alternative is used in the 5 most similar protocols

For surface immunophenotype marker staining, the cells we resuspended in autoMACS^® Rinsing Solution (1 × 10⁵ cells in 100 µL) with 1% BSA and stained with anti-CD16 (A07766, Beckman Coulter), anti-CD14 (130-110-518, Miltenyi Biotec), anti-CD45 (A07785, Beckman Coulter), anti-CD206 (130-095-131, Miltenyi Biotec), anti-CD86 (130-116-160, Miltenyi Biotec), anti-CD163 (130-097-630, Miltenyi Biotec), anti-HLA-DR (130-111-790, Miltenyi Biotec), CD40 (130-110-947, Miltenyi Biotec), CD80 (130-117-683, Miltenyi Biotec) and anti-CX3CR1 (12-6099-42, eBioscience). The analysis was carried out on the FACScan flow cytometer (Becton Dickinson, USA) with the CellQuest software. In each measurement, 10,000 cells were analyzed. The gating strategy included selection of the monocyte population on dot-plot of forward (FSC) and side scatter (SSC), as well as gating on SSC vs CD45 flow dot-plot with further gating on CD14 vs HLA-DR dot-plot to exclude B lymphocytes^{21 (link)}.

Vishnyakova P., Kuznetsova M., Poltavets A., Fomina M., Kiseleva V., Muminova K., Potapova A., Khodzhaeva Z., Pyregov A., Trofimov D., Elchaninov A., Sukhikh G, & Fatkhudinov T. (2022). Distinct gene expression patterns for CD14++ and CD16++ monocytes in preeclampsia. Scientific Reports, 12, 15469.

+ Open protocol

+ Expand

Evaluating Tumor Immune Response to Oncolytic Virus and BiTE

Check if the same lab product or an alternative is used in the 5 most similar protocols

Raji lymphoma cells were implanted intradermally into the right flanks of NSG mice (5 × 10⁶ cells). Mice bearing Raji subcutaneous tumor were treated intratumorally with PBS, OVV, OVV-CD19BiTE (2 × 10⁷ pfu/tumor), or blinatumomab (0.25 mg/kg). Twenty-four hours later, 1 × 10⁷ preactivated T cells were intravenously injected into treated mice. Tumors were harvested 3 days after the injection with forceps and surgical scissors and were weighed. They were then minced before incubation with Liberase (1.67 Wünsch U/mL) and DNase (0.2 mg/mL) in serum-free RPMI for 30 min at 37 °C. Cell suspensions were generated by mashing through a 70 μm nylon filter and then washed with complete RPMI. For tumor-infiltrating T-cells analysis, single-cell suspensions were generated and processed for surface labeling with anti-CD3, anti-CD45, anti-CD4, anti-CD8, anti-HLA-DR, anti-CD45RA, anti-CCR7, anti-CXCR3, and anti-CCR6 antibodies (Beckman). Live cells were distinguished from dead cells by using fixable dye eFluor506 (eBioscience). All flow data were acquired using either an LSRII flow cytometer (BD Biosciences). Data were analyzed with FlowJo software (Treestar).

Lei W., Ye Q., Hao Y., Chen J., Huang Y., Yang L., Wang S, & Qian W. (2022). CD19-targeted BiTE expression by an oncolytic vaccinia virus significantly augments therapeutic efficacy against B-cell lymphoma. Blood Cancer Journal, 12(2), 35.

+ Open protocol

+ Expand

Isolation and Identification of Leukemic Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

PBMCs isolated from bone marrow were stained at room temperature for 15 min in the dark. 7-aminoactinomycon D (7-AAD) and anti-CD45 antibody were used for cells from leukemia patients, 7-AAD and anti-CD19 (all Beckman Coulter) for cells from healthy donors. Sorting was performed in MACS buffer (PBS, 2 mM EDTA, 0.1% BSA) using a FACS Aria (Becton Dickinson). Normal B cells and leukemic blasts were defined as 7-ADD⁻/CD19⁺ and 7-ADD⁻/CD45^low, respectively.

Ziegler N., Cortés-López M., Alt F., Sprang M., Ustjanzew A., Lehmann N., El Malki K., Wingerter A., Russo A., Beck O., Attig S., Roth L., König J., Paret C, & Faber J. (2023). Analysis of RBP expression and binding sites identifies PTBP1 as a regulator of CD19 expression in B-ALL. Oncoimmunology, 12(1), 2184143.

+ Open protocol

+ Expand

Characterization of Human Bone Marrow-Derived Mesenchymal Stem Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Healthy human bone marrow-derived mesenchymal stem cells (BM-MSCs, passage 3) were purchased (Gibco, #A15652) and cultured in low-glucose DMEM (Thermo Fisher Scientific, #11054020) supplemented with 10% Fetal Bovine Serum (FBS - Pan Biotech, #P30-3306), 1% L-glutamine (2 mM, Sigma Aldrich, #G7513) and penicillin/streptomycin antibiotic solution (100 U/0.1 mg/ml, Sigma Aldrich, #P4333) at 37 °C degrees containing 5% CO₂ in humid environment. For the evaluation of cell surface markers, cells were detached with trypsin-EDTA (Thermo Fisher Scientific, #25300054), collected by centrifuging at 300 X G for 5 minutes, and then suspended in Dulbecco’s phosphate buffered saline (DPBS) containing 0.1% sodium azide. Next, the cells were labeled with anti-CD73 FITC (Clone:AD2, Biolegend, #344016), anti-CD90 PE (Clone: 5E10, Biolegend, #328110), anti-CD105 PerCP/Cy5.5 (Clone:43A3, Biolegend, #323216), anti-CD34 PE/Cy7 (Clone:581, Biolegend, #343515), anti-CD11b Alexa Fluor 700 (Clone:ICRF44, Biolegend, #301355), anti-HLA-DR Pacific Blue (Clone:L243, Biolegend, #307633), and anti-CD45 (Clone: J33, Beckman Coulter, #A96416) antibodies by incubating under dark conditions for 15 minutes at room temperature. The cells were immediately acquired on Beckman Coulter DxFLEX flow cytometry system. Analyses were performed on CytExpert software.

Aru B., Akdeniz T., Dağdeviren H., Gürel G, & Yanıkkaya Demirel G. (2023). Testosterone Propionate Promotes Proliferation and Viability of Bone Marrow Mesenchymal Stem Cells while Preserving Their Characteristics and Inducing Their Anti-Cancer Efficacy. Balkan Medical Journal, 40(2), 117-123.

+ Open protocol

+ Expand

Phenotypic Analysis of NK Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

The Monoclonal antibodies (mABs) used for phenotypic analysis included Fluorescein isothiocyanate (FITC)-, Phycoerythrin (PE)-, Phycoerythrin Cyanine 5.5 and allophycocyanin (APC)-labeled anti-CD45, anti-CD3, anti-CD56, NKp30, NKp44 (Beckman Coulter, Brea, CA, USA), anti-CD16, NKp46, NKG2A (R&D System), anti-CD94, anti-CD69, anti-CD18, anti-CD49, anti-CD62L, anti-CD38 and CXCR4 (BD Pharmingen, San Jose, CA, USA). Phenotype evaluation was performed by direct immunofluorescence according to previously reported methods [29 (link)]. NK cells recovered from immunodeficient NOD/SCID gamma (NSG) mice were stained with anti-mouse CD45-APC-Cy7, anti-human CD45-PE-Cy7, CD3-PerCP-Cy5.5, CD56-FITC (Biolegend, San Diego, CA, USA) and CD94-APC (BD).

Tanzi M., Consonni M., Falco M., Ferulli F., Montini E., Pasi A., Cacciatore R., Brugnatelli S., Pedrazzoli P., Zecca M., Boghen S., Dellabona P., Casorati G, & Montagna D. (2021). Cytokine-Induced Memory-Like NK Cells with High Reactivity against Acute Leukemia Blasts and Solid Tumor Cells Suitable for Adoptive Immunotherapy Approaches. Cancers, 13(7), 1577.

+ Open protocol

+ Expand

FACS Sorting of Bone Marrow Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

After thawing and counting the cells from bone marrow samples, cells were stained with 7-aminoactinomycin D (7-AAD) viability dye, anti-CD45, and anti-CD19 (all Beckman Coulter) and incubated at room temperature for 15 minutes. Samples were washed once with 1× PBS (Sigma-Aldrich) and the pellet was resuspended in 500–1000 µL FACS buffer (1× PBS with 2 mM ethylenediaminetetraacetic and 2% fetal calf serum), depending on previously counted cells. FACS sorting was performed with FACS Aria (Becton Dickinson, Heidelberg, Germany). 7-AAD⁻/CD19⁺/CD45^low cells were taken for further analysis.

Fischer J., Paret C., El Malki K., Alt F., Wingerter A., Neu M.A., Kron B., Russo A., Lehmann N., Roth L., Fehr E.M., Attig S., Hohberger A., Kindler T, & Faber J. (2017). CD19 Isoforms Enabling Resistance to CART-19 Immunotherapy Are Expressed in B-ALL Patients at Initial Diagnosis. Journal of Immunotherapy (Hagerstown, Md. : 1997), 40(5), 187-195.

+ Open protocol

+ Expand

Isolation and Characterization of Platelet Microparticles

Check if the same lab product or an alternative is used in the 5 most similar protocols

Peripheral blood was centrifuged at 1000g for 5 min. Plasma was then centrifuged at 1500g for 20 min to obtain platelet poor plasma (PPP). 1 mL of PPP was centrifuged for 30 min at 20,000g to pellet microparticles (MPs). The MP pellet washed once in 500 µL calcium-free HBS complemented with 0.2% bovine serum albumin (pH 7.3) by 45 min centrifugation at 20,000g. MPs were labeled with MitoTracker Deep Red (200 nM; Invitrogen, Breda, The Netherlands) in HBS at room temperature containing calcium and subsequently stained with anti-CD61, anti-CD62p, Annexin-V (all Biolegend), anti-CD45 and anti-CD41 (both Beckman Coulter). MPs were analyzed using a Cytoflex flow cytometer (Beckman Coulter) including the sensitive violet Side Scatter (405 nM; VSSC) and FSC for detection of ultrasmall particles (1 µm)^{54 (link)}. Platelet MPs (PMPs) were selected based on aforementioned markers. Counting beads (Sphero Nano fluorescent, Spherotech, Fulda, Germany) of different sizes were included for reference to correct for concentration and size. All solutions for PMP isolation and staining were centrifuged at 20,000g for 20 min to remove (fluorescent) aggregates. A gating strategy is provided in Supplemental Fig. S2.

van der Heijden W.A., van de Wijer L., Jaeger M., Grintjes K., Netea M.G., Urbanus R.T., van Crevel R., van den Heuvel L.P., Brink M., Rodenburg R.J., de Groot P.G., van der Ven A.J, & de Mast Q. (2021). Long-term treated HIV infection is associated with platelet mitochondrial dysfunction. Scientific Reports, 11, 6246.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!