Log In Sign up
The largest database of trusted experimental protocols

Cd45r b220

Manufactured by BD
Visit Supplier
Sourced in Germany, United States

CD45R/B220 is a cell surface marker that is expressed on B lymphocytes. It is commonly used in flow cytometry for the identification and quantification of B cells.

Automatically generated - may contain errors

Lab products found in correlation

Cd11b, by BD (4 mentions) Ter119, by BD (3 mentions) Ter119, by BioLegend (2 mentions) Lsrfortessa, by BD (2 mentions) Microm hm 450, by Thermo Fisher Scientific (2 mentions) Abc vector elite, by Vector Laboratories (2 mentions) 3 3 diaminobenzidine kit, by Vector Laboratories (2 mentions) Cd138 syndecan 1, by BioLegend (2 mentions) Bz x700 light and fluorescent microscope, by Leica (2 mentions) Sp5 2 confocal laser scanning microscope, by Leica (2 mentions) Alexa fluor secondary antibody, by Thermo Fisher Scientific (2 mentions) Cd16 32, by BD (1 mentions) Il 5rα, by BD (1 mentions) Fix perm, by Thermo Fisher Scientific (1 mentions) Influx flow cytometer, by BD (1 mentions) Percp cy5, by Thermo Fisher Scientific (1 mentions) Efluor 450, by Thermo Fisher Scientific (1 mentions) Fluorescein isothiocyanate (fitc), by Thermo Fisher Scientific (1 mentions) Efluor 780, by Thermo Fisher Scientific (1 mentions) Lsr 2 flow cytometer, by BD (1 mentions)

Close spelling variants of this product

CD45R/B220 biotin (2 citations) APC-labeled anti-mouse CD45R/B220 (4 citations) Anti-CD45R/B220 (11 citations) PE-Rat-Anti-Mouse-CD45R/B220-(RA3-6B2) (2 citations) Anti-CD45R/B220 (clone RA3-6B2; (6 citations) Rat anti-mouse B220/CD45R biotinylated (2 citations) Pacific Blue-labeled rat IgG2aκ anti-mouse CD45R (B220) (clone RA3–6B2; (1 citations) B220/CD45R-APC-Cy7 (1 citations) Anti-CD45R (B220)-FITC (clone RA3-6B2; (2 citations) PE Rat Anti-Mouse CD45R/B220 (5 citations)

26 protocols using cd45r b220

1

Multiparametric Flow Cytometry of Tumor Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
Tumor cell suspensions of representative tumors of each genotype were stained as described30 . The antibodies used were B220/CD45R (BD PharMingen, #553092), IgG1 (BD PharMingen #560089) conjugated with APC, B220/CD45R (BD PharMingen, #553090), CD19 (BD PharMingen, # 557399), IgM (PharMingen, #553409), Thy1/CD90 (Cedarlane, #CL8610PE), CD8 (PharMingen, #553032), Sca-1 (PharMingen, #553108), IgD (BD PharMingen #558597), GL7 (BD PharMingen #561530) conjugated with phycoerythrin and analyzed with BD LSRFortessa™ cell analyzer. Analysis was performed with FlowJo software (Tree Star).
Ortega-Molina A., Boss I.W., Canela A., Pan H., Jiang Y., Zhao C., Jiang M., Hu D., Agirre X., Niesvizky I., Lee J.E., Chen H.T., Ennishi D., Scott D.W., Mottok A., Hother C., Liu S., Cao X.J., Tam W., Shaknovich R., Garcia B.A., Gascoyne R.D., Ge K., Shilatifard A., Elemento O., Nussenzweig A., Melnick A.M, & Wendel H.G. (2015). The histone lysine methyltransferase KMT2D sustains a gene expression program that represses B cell lymphoma development. Nature medicine, 21(10), 1199-1208.
+ Open protocol
+ Expand
2

Analytical Flow Cytometry of Immune Cells

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
All analytical flow cytometry was performed on a BD LSR II flow cytometer. Dead cells were identified by DAPI (4’,6-diamidino-2-phenylindole) and excluded from analysis where appropriate. Primary antibodies specific for the following markers were directly conjugated to FITC, PE, APC, PerCP-Cy5.5, eFluor-780, eFluor-450, or PE-Cy7 and purchased from eBioscience (CD3, CD4, CD8α, CD11b, CD16/32, CD34, CD45.1, CD45.2, CD45R(B220), CD49b(DX5), FcεR1 and Gr1) or from BD Biosciences (c-kit, IL-5Rα, Siglec F, CCR3, and Annexin V). For intracellular staining, surface-labeled cells were fixed in isotonic 3% paraformaldehyde and incubated with affinity-purified anti-CF (Hamilton et al., 2008 (link)) in Fix & Perm (Invitrogen) according to product instructions. Staining was detected with highly cross-adsorbed donkey anti-rabbit-FITC (Jackson Immunochemicals, Stratech, UK). For cell sorting experiments, eosinophil populations were sorted on a BD Influx flow cytometer.
Matthews S.P., McMillan S.J., Colbert J.D., Lawrence R.A, & Watts C. (2016). Cystatin F Ensures Eosinophil Survival by Regulating Granule Biogenesis. Immunity, 44(4), 795-806.
+ Open protocol
+ Expand
3

Comprehensive Phenotyping of PIKfyvefl/fl Pf4-Cre Mice

Check if the same lab product or an alternative is used in the 5 most similar protocols
Necropsies were performed on approximately 50 control mice and 50 PIKfyvefl/fl Pf4-Cre mice between 3 weeks and 28 weeks of age. They included macroscopic observations, body and organ weight measurements, complete blood counts, serum chemistry analysis, and microscopic examination of the brain, thymus, lymph nodes, heart, lung, small intestine, large intestine, liver, spleen, pancreas, kidney, ovaries, testis, skin, muscle, fat, and bone marrow. Fresh postmortem tissues were fixed overnight in 10% formalin, paraffin-embedded, sectioned, and stained with hematoxylin/eosin, PAS, Alcian Blue, and Trichrome Mason. Frozen tissue sections were fixed with formalin for 5 minutes and stained for oil red O. For immunohistochemistry, frozen tissue sections were fixed with 4% paraformaldehyde for 5 minutes, and stained with F4/80 (Serotec; 1:100), CD41 (BD Pharmingen; MWReg30; 1:50), and CD45R/B220 (BD Pharmingen; RA3–6B2; 1:500). For X-gal staining, frozen tissue sections were fixed with 0.5% glutaraldehyde for 10 minutes, and stained overnight with the chromogenic substrate for β-galactosidase, X-gal (Cell Center, University of Pennsylvania).
Min S.H., Suzuki A., Stalker T.J., Zhao L., Wang Y., McKennan C., Riese M.J., Guzman J.F., Zhang S., Lian L., Joshi R., Meng R., Seeholzer S.H., Choi J.K., Koretzky G., Marks M.S, & Abrams C.S. (2014). Loss of PIKfyve in platelets causes a lysosomal disease leading to inflammation and thrombosis in mice. Nature communications, 5, 4691.
+ Open protocol
+ Expand
4

Wnt Signaling Activation in Hematopoietic Stem Cells

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Mononuclear BM cells from WT or Fancc−/− mice (CD45.2) were labeled with the lineage marker (Lin) biotinylated Abs mixture (BD Biosciences) including: anti- CD3ε (Clone 145-2C11), CD11b (Clone M1/70), CD45R/B220 (Clone RA3-6B2), Ly-76 (Clone Ter119), Ly6G, and Ly-6C (Clone RB6-8C5). Cells were then labeled with anti-Sca1-PeCy7 (D7) and anti-cKit-APC (2B8) (BD Biosciences). After washing, biotinylated Abs were revealed by Streptavidin Percp-Cy5.5 (BD Biosciences). Lin−Sca1+Ckit+ (LSK) cells were sorted using FACS Aria (BD Biosciences) from the CCHMC Flow Cytometry Core. Sorted LSKs (≥98%) were activated in StemSpan medium (Stemcell technologies) in presence of 25ng/ml murine rTpo (Preprotech) and 50ng/ml murine rScf (Preprotech). After 12h, LSKs were transduced by the 7TGC-eGFP Wnt reporter lentivirus (23 (link)) with 2 hits at MOI=5. Lentiviral particles were produced by the CCHMC Viral Vector core using 293T cells. At 72h, transduction efficiency was evaluated by mCherry detection by Flow cytometry (Transduction efficiency 70-80%). Five thousand LSK cells were transplanted into lethally irradiated Boy/J recipient mice (CD45.1). GFP expression of naïve splenic B cells was detected at 10 weeks post transplantation by Flow cytometry.
Sertorio M., Amarachintha S., Wilson A, & Pang Q. (2016). Loss of Fancc impairs antibody secreting cell differentiation in mice through deregulating Wnt signaling pathway. Journal of immunology (Baltimore, Md. : 1950), 196(7), 2986-2994.
+ Open protocol
+ Expand
5

Flow Cytometry Protocol for Murine Immune Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
Flow cytometry analysis was performed as described (35 (link)). The BD LSR Fortessa (BD Biosciences, Heidelberg, Germany) was used for analysis. Antibodies used to stain cell surface proteins were anti-mouse CD4 (GK1.5), CD8a (53-6.7), CD11b (Mac-1, M1/70), CD25 (PC61.5), CD44 (IM7), CD45R/B220 (RA3-6B2), CD45 (30-F11), CD90.2 (THY1.2, 53-2.1), CD117 (c-KIT, 2B8), CD127 (IL-7Ra, A7R34), GR1 (Ly-6G, RB6-8C5), SCA1 (D7), and TER119 (TER119) and the corresponding isotypes, which were obtained from BD Biosciences or eBiosciences (Frankfurt am Main, Germany).
Kreutmair S., Lippert L.J., Klingeberg C., Albers-Leischner C., Yacob S., Shlyakhto V., Mueller T., Mueller-Rudorf A., Yu C., Gorantla S.P., Miething C., Duyster J, & Illert A.L. (2022). NIPA (Nuclear Interaction Partner of ALK) Is Crucial for Effective NPM-ALK Mediated Lymphomagenesis. Frontiers in Oncology, 12, 875117.
+ Open protocol
+ Expand
6

Comprehensive Flow Cytometry Immunophenotyping

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were stained using Abs specific for mouse Ags: CD45R (B220) (various fluorochromes) (BD Pharmingen, BioLegend, San Diego, CA and Tonbo Biosciences); IgM (various fluorochromes, Jackson ImmuoResearch Laboratories, West Grove, PA); CD19 eFlour450, CD25 APC, MHC II APC (Tonbo Biosciences); CD43 PE, BP-1 PE, CD117 PE, CD24 (HSA) PE-Cy7, IgD FITC (BD Pharmingen); CD21 PerCP/Cy5.5 (BioLegend). IC staining was performed with IC fixation and permeabilization buffer (eBiosciences). Methanol fixation was performed for IC phospho (p)-S6R detection. Abs used for IC staining were p-ribosomal S6 protein (S6R) S235/236 PE (eBiosciences); p-AMPKα T172, c-Myc AF488 and p-Ulk1 S555 (Cell Signaling Technology, Danvers, MA). Donkey anti-rabbit Alexa-Fluor 647 (Life Technologies, Carlsbad CA) secondary Ab was used to detect unlabeled primary Abs. Data was collected using FACS Canto II or LSR II flow cytometers (BD Biosciences) and analyses were performed using FlowJo software (TreeStar, Ashland, OR).
Ramίrez J.A., Iwata T., Park H., Tsang M., Kang J., Cui K., Kwong W., James R.G., Baba M., Schmidt L.S, & Iritani B.M. (2019). Folliculin Interacting Protein-1 Maintains Metabolic Homeostasis During B Cell Development by Modulating AMPK, mTORC1, and TFE3. Journal of immunology (Baltimore, Md. : 1950), 203(11), 2899-2908.
+ Open protocol
+ Expand
7

Multiparametric Immune Cell Profiling

Check if the same lab product or an alternative is used in the 5 most similar protocols
Blood cells were collected from the eye vein of anesthetized mice, and splenocytes from the spleen after filtration through cell strainers (70 µm). Ammonium-chloride-potassium lysing buffer was used for the lysis of red blood cells. Blood cells were labeled for 15 min at 4°C with fluorochrome-conjugated monoclonal antibodies: CD45-FITC BD Biosciences Clone 30-F11 (1:100), Ly6G BV421 BioLegend Clone 1A8 (1:500), Ly6C BV605 BD Biosciences Clone AL-21 (1:500), CD3e BV786 BD Biosciences Clone 5OOA2 (1:500), CD45R (B220) PE BD Biosciences Clone RA3-6B2 (1:500) and CD11b PE-Cy7 eBioscience Clone M1/70 (1:500). Splenocytes were labeled for 15 min at 4°C with fluorochrome-conjugated monoclonal antibodies: CD3 BV786 BD Biosciences (1:500), CD11b BV510 BD Biosciences (1:500), CD49b PE eBioscience (1:500), CD4 BV650 BD Biosciences (1:500), CD25 FITC BioLegend (1:500) and Foxp3 PerCP-Cy5.5 eBioscience (1:100). Samples were acquired with a BD LSRII Fortessa (BD Biosciences) and analyzed with FlowJo software.
Cordero-Sanchez C., Riva B., Reano S., Clemente N., Zaggia I., Ruffinatti F.A., Potenzieri A., Pirali T., Raffa S., Sangaletti S., Colombo M.P., Bertoni A., Garibaldi M., Filigheddu N, & Genazzani A.A. (2019). A luminal EF-hand mutation in STIM1 in mice causes the clinical hallmarks of tubular aggregate myopathy. Disease Models & Mechanisms, 13(2), dmm041111.
+ Open protocol
+ Expand
8

Immunohistochemical Analysis of Lung Tissue

Check if the same lab product or an alternative is used in the 5 most similar protocols
For immunohistochemistry, lungs were fixed in paraformaldehyde and embedded into paraffin. After deparaffinizing in xylene and rehydrating in alcohol, the tissue was treated with 1.8% (v/v) H2O2 solution (Sigma-Aldrich, St. Louis, MO) to block endogenous peroxidase. Heat induced epitope retrieval was performed in HIER Citrate Buffer (pH 6.0, Zytomed Systems) in a Decloaking chamber (Biocare Medical, Concord, CA). To inhibit nonspecific binding of antibodies, tissue slides were treated with a rodent blocking antibody (Biocare Medical). After overnight incubation with primary antibodies against MMP12 (Abcam, Cambridge, UK), CD45R/B220 (BD Pharmingen), CD21 (Novus Biologicals, Littleton, CO), CD3 (Sigma Aldrich), p16 (Santa Cruz, CA) or SIRT1 (Millipore, Schwalbach, Germany) tissue slides were incubated with an alkaline phosphatase-labeled secondary antibody (Biocare Medical). Signals were amplified by adding chromogen substrate Vulcan fast red (Biocare Medical). Slides were counterstained with hematoxylin (Sigma-Aldrich) and dehydrated in xylene. Afterwards, coverslips were mounted.
John-Schuster G., Günter S., Hager K., Conlon T.M., Eickelberg O, & Yildirim A.Ö. (2015). Inflammaging increases susceptibility to cigarette smoke-induced COPD. Oncotarget, 7(21), 30068-30083.
+ Open protocol
+ Expand
9

Histological Analysis of Mouse Lung Tissue

Check if the same lab product or an alternative is used in the 5 most similar protocols
Lungs were removed and immersion fixed for 24 h with 4% buffered formaldehyde solution pH 7.0 (Merck KGaA). After embedding in paraffin, tissue sections were cut with a microtome, mounted onto glass slides, and stained.
For histopathological analyses of naïve mice, 3-µm sections were either stained with hematoxylin and eosin (H&E) or immunostained for CD45R/B220 (B cells, monoclonal, 1:1,000, clone RA3-6B2, 553086; BD Biosciences, Heidelberg, Germany) or CD3 (T cells, polyclonal, 1:800; reference A0452; Dako, Santa Clara, CA, USA). Positive immunostaining was visualized using diaminobenzidine, and slides were counterstained with hematoxylin. For analyses of the effects of pulmonary Th2 inflammation, 5-µm tissue sections were either stained with H&E or Masson–Goldner trichrome.
Microscopic analyses were performed (Axiophot; Carl Zeiss Microscopy GmbH, Jena, Germany) in a blinded fashion by a board-certified veterinary pathologist or an anatomist and images were digitized (Color View II camera, CellSens software; Olympus Europa SE Co. KG, Hamburg, Germany).
Tabeling C., González Calera C.R., Lienau J., Höppner J., Tschernig T., Kershaw O., Gutbier B., Naujoks J., Herbert J., Opitz B., Gruber A.D., Hocher B., Suttorp N., Heidecke H., Burmester G.R., Riemekasten G., Siegert E., Kuebler W.M, & Witzenrath M. (2022). Endothelin B Receptor Immunodynamics in Pulmonary Arterial Hypertension. Frontiers in Immunology, 13, 895501.
+ Open protocol
+ Expand
10

Murine Immune Cell Isolation Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols
Adult female, 6–8 week old C3H/HeN mice were obtained from Charles River Laboratories (Wilmington, MA). Animals were maintained in accordance with institutional guidelines. Normal goat IgG and goat polyclonal anti-HSP27 and anti-HSP70 IgG were purchased from Santa Cruz Biotechnology CA, USA. Sheep anti-rat IgG dynabeads were from Life Technologies (Carlsbad,CA). Hybridoma lines GK1.5 (anti-CD4), Lyt-2 (anti-CD8) and HB-32 (anti-Iak) were acquired from ATCC (Manassas, VA). CD45R/B220 and recombinant GM-CSF were obtained from Pharmingen (San Diego, CA). DMBA, TPA, benzo(a)pyrene (B(a)P), and recombinant IL-4 and were purchased from Sigma Chemical Co. (St. Louis, MO).
Yusuf N., Nasti T.H., Ahmad I., Chowdhury S., Mohiuddin H., Xu H., Athar M., Timares L, & Elmets C.A. (2015). In vivo suppression of HSP27 and HSP70 accelerates DMBA-induced skin carcinogenesis by inducing antigenic unresponsiveness to the initiating carcinogenic chemical. Journal of immunology (Baltimore, Md. : 1950), 194(10), 4796-4803.
+ 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!