Log In Sign up
The largest database of trusted experimental protocols

Cd45ra apc h7

Manufactured by BD
Visit Supplier
Sourced in United States

CD45RA-APC-H7 is a fluorescently-labeled monoclonal antibody that binds to the CD45RA isoform of the CD45 protein, which is expressed on the surface of certain types of immune cells. This product is intended for use in flow cytometry applications to identify and quantify CD45RA-positive cells within a sample.

Automatically generated - may contain errors

Lab products found in correlation

Cd8 percp, by BD (2 mentions) Cd4 apc, by BD (2 mentions) Cd19 bv510, by BioLegend (2 mentions) Facscanto 2, by BD (2 mentions) Cd69 apc, by BD (1 mentions) Cd45ro pe, by BD (1 mentions) Cytotox 96 non radioactive cytotoxicity assay, by Promega (1 mentions) Anti cd3 fitc, by BD (1 mentions) Live dead fixable aqua, by Thermo Fisher Scientific (1 mentions) Ccr7 pe cf594, by BD (1 mentions) Cd8 bv711, by BD (1 mentions) Cd14 bv510, by BD (1 mentions) Cd57 pe, by BioLegend (1 mentions) Pd 1 bv605, by BioLegend (1 mentions) Cd69 bv650, by BioLegend (1 mentions) Cd38 bv421, by BioLegend (1 mentions) Tigit pe cy7, by BioLegend (1 mentions) Dnam bb515, by BD (1 mentions) Caspase 3 7 green flow cytometry assay kit, by Thermo Fisher Scientific (1 mentions) Cd4 buv395, by BD (1 mentions)

Close spelling variants of this product

CD45RA-APC (16 citations) Anti-CD45RA-APC-H7 (6 citations) CD45RA APC-H7 (clone HI100; (3 citations) APC-H7 anti-CD45RA (cloneHI100; (2 citations) Anti‐CD45RA‐APC‐H7 (HI100 (2 citations) CD45RA APC-H7 clone 5H9 (2 citations)

12 protocols using cd45ra apc h7

1

Characterization of Human and Rhesus HSCs

Check if the same lab product or an alternative is used in the 5 most similar protocols
Freshly thawed human and rhesus CD34+ cells were incubated with human Fc Block™ (Becton Dickinson (BD), Franklin Lakes, NJ, USA, 564219) for 10 minutes at room temperature (RT) to minimize non-specific antibody interactions. These cells were then subjected to a 30-minute staining procedure on ice (all antibodies used in 1:25 dilution), utilizing two distinct antibody panels with or without PE-anti-cMPL (Miltenyi Biotech) or PE-anti-cKIT (BioLegend, San Diego, CA, USA, 105807). The human HSC panel included FITC-anti-CD34 (BD, 560942), APC-anti-CD38 (BD, 340439), PE-Cy7-anti-CD90 (BD, 561558), APC-H7-CD45RA (BD, 560674), and PE-Cy5-anti-CD49f (BD, 551129) antibodies. The rhesus HSC panel comprised BV421-anti-CD34 (BD, 740081), APC-anti-CD38 (NHP Resources), PE-Cy7-anti-CD90 (BD, 561558), APC-H7-CD45RA (BD, 561212) and PE-Cy5-anti-CD49f (BD, 551129) antibodies. In a specified set of experiments, antibodies underwent conjugation employing a PE conjugation lightning-link kit (Abcam, Waltham, MA, USA, ab102918). A detailed inventory of the antibodies utilized is available in Key Resources Table. Stained cells were subjected to a single wash with FACS buffer and then filtered through 40 μm cell strainers. Quantitative and qualitative analyses of surface marker expression were then conducted using an LSR II Fortessa flow cytometer (BD).
Araki D., Hong S., Linde N., Fisk B., Redekar N., Salisbury-Ruf C., Krouse A., Engels T., Golomb J., Dagur P., Magnani D.M., Wang Z, & Larochelle A. (2024). cMPL-Based Purification and Depletion of Human Hematopoietic Stem Cells: Implications for Pre-Transplant Conditioning. bioRxiv.
+ Open protocol
+ Expand
2

T-cell Stimulation and Cytotoxicity Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
On day 0, 7, and 14, the PBMCs from the T‐cell stimulation assay were analyzed in flow‐cytometry by anti‐CD3‐FITC, CD8‐PerCP, CD45RO‐PE, CD45RA‐APC‐H7, and CD69‐APC antibodies (BD, Heidelberg, Germany) with a FACS Canto II.
Cytotoxicity of effector cells that were stimulated for 14 days was tested by measuring LDH release with the CytoTox 96 Non‐Radioactive Cytotoxicity Assay (Promega, Madison, Wisconsin) following incubation with peptide loaded T2/mHLA‐A*24:02 cells at a ratio of 5:1. The cells were incubated for 4 hours at 37°C and LDH release was measured according to kit instructions.
Pump W.C., Schulz R., Huyton T., Kunze‐Schumacher H., Martens J., Hò G.T., Blasczyk R, & Bade‐Doeding C. (2019). Releasing the concept of HLA‐allele specific peptide anchors in viral infections: A non‐canonical naturally presented human cytomegalovirus‐derived HLA‐A*24:02 restricted peptide drives exquisite immunogenicity. Hla, 94(1), 25-38.
+ Open protocol
+ Expand
3

Multiparameter Flow Cytometry Phenotyping

Check if the same lab product or an alternative is used in the 5 most similar protocols
Standard extracellular staining (20 mins at 24°C) was performed using the following fluorophore-labeled antibodies: CD3-BUV737 (BD Biosciences), CD4-BUV395 (BD Biosciences), CD8-BV711 (BD Biosciences), CD14-BV510 (BD Biosciences), CD19-BV510 (BioLegend), Live/Dead Fixable Aqua (ThermoFisher), CCR7-PE-CF594 (BD Biosciences), CD45RA-APC-H7 (BD Biosciences), CD28-APC-R700 (BD Biosciences), CD57-PE (BioLegend), PD-1-BV605, BV421 (BioLegend), CD69-BV650 (BioLegend), CD38-BV421 (BioLegend), HLA-DR-PerCP-Cy5.5 (BD Biosciences), TIGIT-PE-Cy7 (BioLegend), DNAM-BB515 (BD Biosciences), FcγRIIB-BV786 (BD Biosciences). Fluorescence minus one (FMO) controls were used to determine negative expression of FcγRIIB. Apoptosis was measured with Caspase-3/7 Green Flow Cytometry Assay Kit (ThermoFisher) or FITC Annexin V with 7-AAD Viability Staining Solution (BioLegend), according to the manufacturer’s instructions.
Sun H., Hartigan C.R., Chen C.W., Sun Y., Tariq M., Robertson J.M., Krummey S.M., Mehta A.K, & Ford M.L. (2021). TIGIT regulates apoptosis of risky memory T cell subsets implicated in belatacept-resistant rejection. American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons, 21(10), 3256-3267.
+ Open protocol
+ Expand
4

Multicolor Flow Cytometry Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cell surface marker expression analysis was conducted by flow cytometry using either a FACSCalibur (for 3-color panels, Becton Dickinson, East Rutherford, NJ) or a BD FACSCanto flow cytometer (for 6-color panels) after fluorescent antibody labeling. All antibodies were provided by BD Biosciences (San Jose, CA) as follows: CD31-APC Cy7 (clone WM59), CD31-FITC (WM59), CD34-PErCP Cy5.5 (clone 8G12), CD34-PE (clone 563), CD34-FITC (clone 581), CD38-APC (Clone HIT2), CD43-APC (clone 1G10), CD45-APC (Clone HI30), CD45-APC Cy7 (Clone 2D1), CD45RA-APC H7 (Clone HI100), CD49f-PE (Clone GoH3), CD73-APC (clone M-A712), CD90-PE Cy7 (Clone 5E10), CD144-FITC (Clone 55-7H1), CD235a-PE Cy7 (clone GA-R2), CD235a-PE (clone GA-R2), and DLL4-PE (Clone MHD4-46). Sorting experiments were performed using a BD FACSAria II instrument.
Demirci S., Haro-Mora J.J., Leonard A., Drysdale C., Malide D., Keyvanfar K., Essawi K., Vizcardo R., Tamaoki N., Restifo N.P., Tisdale J.F, & Uchida N. (2020). Definitive hematopoietic stem/progenitor cells from human embryonic stem cells through serum/feeder-free organoid-induced differentiation. Stem Cell Research & Therapy, 11, 493.
+ Open protocol
+ Expand
5

Comprehensive B and T Cell Immunophenotyping

Check if the same lab product or an alternative is used in the 5 most similar protocols
PB was collected in heparin-coated tubes (Venosafe plastic tubes, Terumo Europe N.V., Leuven, Belgium) and PB mononuclear cells (PBMC) were isolated using high density centrifugation (Lympholyte; Cedarlane Laboratories, SanBio B.V., Uden, the Netherlands). PBMC (0.5×106 cells) were stained using anti-human CD19 PerCP-Cy5.5 and CD4 APC to discriminate between B and T cells, respectively (BD Biosciences, Erembodegem, Belgium). B cell subpopulations and surface molecules were defined using following anti-human antibodies: IgD APC-Cy7, CD27 PE-Cy7, HLA-DR/DP/DQ (major histocompatibility complex (MHC)-II) FITC, CD80 PE and CD86 PE-CF594 (all from BD Biosciences, Erembodegem, Belgium). Following anti-human monoclonal antibodies were used for T cell analysis: CD45RA APC-H7, CD45RO PE-CF594, CXCR5 Alexa Fluor 488 and PD-1 PE-Cy7 (all from BD Biosciences, Erembodegem, Belgium), CD25 PerCP-Cy5.5 and CD127 PE (eBioscience, San Diego, USA). Following isotype controls were used: mouse IgG1 PErCP-Cy5.5, IgG1 PE, IgG1 PE-Cy7, IgG2aκ PE-CF594, IgG2bκ APC-H7, IgG1 APC, IgG2aκ FITC, IgG1κ PE-CF594, IgG1 PE-Cy7, IgG2aκ APC-H7 and rat IgG2b Alexa Fluor 488 (all from BD Biosciences, Erembodegem, Belgium). All flow cytometric analyses were performed on a FACSAriaII flow cytometer and analyzed with FACS Diva software (BD Biosciences).
Claes N., Dhaeze T., Fraussen J., Broux B., Van Wijmeersch B., Stinissen P., Hupperts R., Hellings N, & Somers V. (2014). Compositional Changes of B and T Cell Subtypes during Fingolimod Treatment in Multiple Sclerosis Patients: A 12-Month Follow-Up Study. PLoS ONE, 9(10), e111115.
+ Open protocol
+ Expand
6

PBMC Isolation, Sorting, and RNA-Seq

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Cryopreserved PBMC were thawed in media containing 20% fetal bovine serum. Cell counts and viabilities were assessed using Guava ViaCount reagent and a Guava PCA (Millipore Sigma). Cells were washed and stained with Aqua Live/Dead stain (Molecular Probes), washed, and blocked using normal mouse IgG (Caltag). The cells were surface stained for CD3 FITC (clone UCHT1, Becton Dickinson [BD]), CD8 PerCP-eF710 (SK1, eBiosciences), CD14 V500 (M5E2, BD), CD45RO eF650NC (UCHL1, eBiosciences), CD4 APC (RPAT4, BD), CD45RA APC-H7 (HI100, BD), CD19 PE-Cy5 (SJ25-C1, Invitrogen), and CD56 PE-Cy7 (NCAM16.2, BD) (Figure S2). The cells were then washed and sorted/analyzed on the BD FACSAria II SORP Cell Sorter. Total RNA from sorted cell subsets was extracted using the Single Cell RNA Purification kit (Norgen Biotek Corp). RNA was prepared for NGS RNA sequencing (RNAseq) using the SMART-Seq technology as described previously (Ehrenberg et al., 2019 ; Picelli et al., 2014 (link); Ramsköld et al., 2012 (link); Shangguan et al., 2021 ). Briefly, cDNA was generated per manufacturer’s instructions from 2.5 ng of RNA, final exogenous ERCC RNA spike-in dilutions of 1:190,000, and 11 cycles of library PCR amplification. A total of 300 ng of cDNA was input template for final library processing using the Nextera XT kit (Illumina) per manufacturer’s instructions.
Li S.S., Hickey A., Shangguan S., Ehrenberg P.K., Geretz A., Butler L., Kundu G., Apps R., Creegan M., Clifford R.J., Pinyakorn S., Eller L.A., Luechai P., Gilbert P.B., Holtz T.H., Chitwarakorn A., Sacdalan C., Kroon E., Phanuphak N., de Souza M., Ananworanich J., O’Connell R.J., Robb M.L., Michael N.L., Vasan S, & Thomas R. (2022). HLA-B*46 associates with rapid HIV disease progression in Asian cohorts and prominent differences in NK cell phenotype. Cell host & microbe, 30(8), 1173-1185.e8.
+ Open protocol
+ Expand
7

Targeted single-cell RNA-seq analysis of CD4+ T cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
Fresh whole blood was collected from patients and HDs. PBMCs were isolated by density gradient centrifugation. Leukocytes were labeled with Aqua LIVE/DEAD (Thermo Fisher Scientific) stain for the removal of non-viable cells, and with the following antibodies: CD45RA-APC-H7, CD3-BV650 and CD4-R718 (BD Biosciences). CD45RA-CD3+CD4+ TLs were isolated from PBMCs by flow cytometry sorting. Targeted scRNA-seq analysis was performed with the BD Rhapsody Single-Cell Analysis System (BD Biosciences), according to the manufacturer’s instructions. Each sample was stained with 20 BD AbSeq antibodies (Supplemental Table 5). The BD Human Single-Cell Multiplexing Kit was used to multiplex up to seven samples per Rhapsody cartridge. For library construction, the samples were pooled before cartridge loading. The BD Rhapsody Immune Response Targeted Panel for Humans was used to assess mRNA levels for 399 genes (#633750).
Khelfa M., Leclerc M., Kerbrat S., Boudjemai Y.N., Benchouaia M., Neyrinck-Leglantier D., Cagnet L., Berradhia L., Tamagne M., Croisille L., Pirenne F., Maury S, & Vingert B. (2023). Divergent CD4+ T-cell profiles are associated with anti-HLA alloimmunization status in platelet-transfused AML patients. Frontiers in Immunology, 14, 1165973.
+ Open protocol
+ Expand
8

CD8+ T Cell Phenotypic Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
After 10 days of co-culture, the CD8+ T cells were phenotypically analyzed by flow cytometry. Cells were stained with anti- CD3-PE-Cy7, CD8-PerCP, CD56-FITC, CD45RA-APC-H7 and CD45RO-PE-Cy7 mAb (all from BD Biosciences). Furthermore, we used anti-NKG2A-APC and anti-NKG2C-PE mAb (R&D systems, Minneapolis, MN, USA). Live/Dead discrimination was performed by 7-AAD staining (Biolegend, San Diego, CA, USA) according to manufacturer protocol (Supplementary Figure S5). All samples were analyzed on a FACS Canto II (BD Biosciences).
Pump W.C., Kraemer T., Huyton T., Hò G.G., Blasczyk R, & Bade-Doeding C. (2019). Between Innate and Adaptive Immune Responses: NKG2A, NKG2C, and CD8+ T Cell Recognition of HLA-E Restricted Self-Peptides Acquired in the Absence of HLA-Ia. International Journal of Molecular Sciences, 20(6), 1454.
+ Open protocol
+ Expand
9

Multiparametric Flow Cytometry Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were stained with antibodies against: anti-human CD3-PERCP (BD), anti-human gp91 phox -FITC (anti-flavocytochrome B558, clone 7D5) (MBL, Medical and Biological Laboratories Co., Japan), CD45RA-APCH7 (BD), CD4-APC (BD), CD8-APC (BD). Intracellular staining was performed using cytofix-cytoperm (BD) in accordance with the manufacturer's instructions. FACS Canto II (Becton-Dickinson, USA) and Flowjo (Tree Star, Inc.) were used to collect and analyze the data obtained.
Chiriaco M., Casciano F., Di Matteo G., Gentner B., Claps A., Di Cesare S., Cotugno N., D'Argenio P., Rossi P., Aiuti A, & Finocchi A. (2018). Impaired X-CGD T cell compartment is gp91phox-NADPH oxidase independent. Clinical immunology (Orlando, Fla.), 193.
+ Open protocol
+ Expand
10

Isolation and Analysis of Fallopian Tube Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
Fresh fragments of human fallopian tubes were initially washed with saline solution to eliminate the large majority of circulating mononuclear cells. Then they were finely dissected, minced and digested using 1 mg/mL collagenase IV (Sigma–Aldrich, Dorset, UK) in PBS for 2 h at 37 °C. Digested tissue was filtered through 70 μm and 40 μm filters. Filtered material containing the fallopian tube single cell suspension was centrifuged twice for 10 min at 1500 rpm and stained with a mixture of the following monoclonal antibodies (mABs): iNKT-FITC, CD16-PE, CD8-PerCP-Cy5.5, CD3-PE-Cy7, CD45RA-APC-H7, CD4-V450 and CD45-V500 (all from Becton Dickinson, Franklin Lakes, NJ, USA) and CD56-APC (Miltenyi Biotec, Bergisch Gladbach, Germany). At least 50000 lymphocytes were acquired from each sample using a FACSCanto II system (Becton Dickinson), while analyses were performed with FlowJo software version 8.8.7 (TreeStar, Ashland, OR, USA).
Ardighieri L., Lonardi S., Moratto D., Facchetti F., Shih I.M., Vermi W, & Kurman R.J. (2014). Characterization of the Immune Cell Repertoire in the Normal Fallopian Tube. International journal of gynecological pathology : official journal of the International Society of Gynecological Pathologists, 33(6), 581-591.
+ 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?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!