50-0452-80

antibody from Invitrogen Antibodies

Targeting: PTPRC CD45, GP180, LCA, T200

Immunocytochemistry (Recommended by provider)

Immunohistochemistry (Data presented on provider website)

Flow cytometry (Supportive data in Antibodypedia)

Antibody Data

Product number

50-0452-80

Provider

Invitrogen Antibodies

Product name

Anti-CD45R (B220) Monoclonal Antibody (RA3-6B2), eFluor 660, eBioscience™

Antibody type

Monoclonal

Antigen

Other

Description

Description: The RA3-6B2 monoclonal antibody reacts with exon A-restricted isoform of mouse CD45, a 220 kDa surface molecule. CD45R/B220 epitope is mainly expressed by the B cell lineage from early Pro-B to mature B cells. However, some activated T cells, lymphokine activated killer cells (LAK), NK cell progenitors in the bone marrow, and T cells of the \I{lpr/lpr\I} mutant mouse also express this antigen. Applications Reported: This RA3-6B2 antibody has been reported for use in flow cytometric analysis, and immunohistochemical staining of frozen tissue sections. Applications Tested: This RA3-6B2 antibody has been tested by flow cytometric analysis of mouse splenocytes. This can be used at less than or equal to 0.25 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. This RA3-6B2 antibody has been tested by immunohistochemistry on frozen mouse spleen and can be used at less than or equal to 10 µg/mL. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. eFluor® 660 is a replacement for Alexa Fluor® 647. eFluor® 660 emits at 659 nm and is excited with the red laser (633 nm). Please make sure that your instrument is capable of detecting this fluorochome. Excitation: 633-647 nm; Emission: 668 nm; Laser: Red Laser. Filtration: 0.2 µm post-manufacturing filtered.

Reactivity

Human, Mouse

Host

Rat

Isotype

IgG

Antibody clone number

RA3-6B2

Vial size

25 µg

Concentration

0.2 mg/mL

Storage

4° C, store in dark, DO NOT FREEZE!

References

Submitted references

Tpl2 Protects Against Fulminant Hepatitis Through Mobilization of Myeloid-Derived Suppressor Cells.

Xu J, Pei S, Wang Y, Liu J, Qian Y, Huang M, Zhang Y, Xiao Y
Frontiers in immunology 2019;10:1980

---

Anti-commensal IgG Drives Intestinal Inflammation and Type 17 Immunity in Ulcerative Colitis.

Castro-Dopico T, Dennison TW, Ferdinand JR, Mathews RJ, Fleming A, Clift D, Stewart BJ, Jing C, Strongili K, Labzin LI, Monk EJM, Saeb-Parsy K, Bryant CE, Clare S, Parkes M, Clatworthy MR
Immunity 2019 Apr 16;50(4):1099-1114.e10

---

Proapoptotic BIM Impacts B Lymphoid Homeostasis by Limiting the Survival of Mature B Cells in a Cell-Autonomous Manner.

Liu R, King A, Bouillet P, Tarlinton DM, Strasser A, Heierhorst J
Frontiers in immunology 2018;9:592

---

ATF3 Sustains IL-22-Induced STAT3 Phosphorylation to Maintain Mucosal Immunity Through Inhibiting Phosphatases.

Glal D, Sudhakar JN, Lu HH, Liu MC, Chiang HY, Liu YC, Cheng CF, Shui JW
Frontiers in immunology 2018;9:2522

---

Microbial symbionts regulate the primary Ig repertoire.

Chen Y, Chaudhary N, Yang N, Granato A, Turner JA, Howard SL, Devereaux C, Zuo T, Shrestha A, Goel RR, Neuberg D, Wesemann DR
The Journal of experimental medicine 2018 May 7;215(5):1397-1415

---

Host Tumor Suppressor p18^INK4c Functions as a Potent Cell-Intrinsic Inhibitor of Murine Gammaherpesvirus 68 Reactivation and Pathogenesis.

Niemeyer BF, Oko LM, Medina EM, Oldenburg DG, White DW, Cool CD, Clambey ET, van Dyk LF
Journal of virology 2018 Mar 15;92(6)

---

Adrenergic nerve degeneration in bone marrow drives aging of the hematopoietic stem cell niche.

Maryanovich M, Zahalka AH, Pierce H, Pinho S, Nakahara F, Asada N, Wei Q, Wang X, Ciero P, Xu J, Leftin A, Frenette PS
Nature medicine 2018 Jun;24(6):782-791

---

Anti-inflammatory Effects of the Octapeptide NAP in Human Microbiota-Associated Mice Suffering from Subacute Ileitis.

Escher U, Giladi E, Dunay IR, Bereswill S, Gozes I, Heimesaat MM
European journal of microbiology & immunology 2018 Jun 25;8(2):34-40

---

miR-143/145 differentially regulate hematopoietic stem and progenitor activity through suppression of canonical TGFβ signaling.

Lam J, van den Bosch M, Wegrzyn J, Parker J, Ibrahim R, Slowski K, Chang L, Martinez-Høyer S, Condorelli G, Boldin M, Deng Y, Umlandt P, Fuller M, Karsan A
Nature communications 2018 Jun 20;9(1):2418

---

Oxysterol Sensing through the Receptor GPR183 Promotes the Lymphoid-Tissue-Inducing Function of Innate Lymphoid Cells and Colonic Inflammation.

Emgård J, Kammoun H, García-Cassani B, Chesné J, Parigi SM, Jacob JM, Cheng HW, Evren E, Das S, Czarnewski P, Sleiers N, Melo-Gonzalez F, Kvedaraite E, Svensson M, Scandella E, Hepworth MR, Huber S, Ludewig B, Peduto L, Villablanca EJ, Veiga-Fernandes H, Pereira JP, Flavell RA, Willinger T
Immunity 2018 Jan 16;48(1):120-132.e8

---

Tanshinone IIA and Astragaloside IV promote the angiogenesis of mesenchymal stem cell-derived endothelial cell-like cells via upregulation of Cx37, Cx40 and Cx43.

Li Z, Zhang S, Cao L, Li W, Ye YC, Shi ZX, Wang ZR, Sun LX, Wang JW, Jia LT, Wang W
Experimental and therapeutic medicine 2018 Feb;15(2):1847-1854

---

Inactivation of mTORC1 Signaling in Osterix-Expressing Cells Impairs B-cell Differentiation.

Wang Y, Xiao M, Tao C, Chen J, Wang Z, Yang J, Chen Z, Zou Z, Liu A, Cai D, Jiang Y, Ding C, Li M, Bai X
Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research 2018 Apr;33(4):732-742

---

The Interleukin (IL)-1R1 pathway is a critical negative regulator of PyMT-mediated mammary tumorigenesis and pulmonary metastasis.

Dagenais M, Dupaul-Chicoine J, Douglas T, Champagne C, Morizot A, Saleh M
Oncoimmunology 2017;6(3):e1287247

---

Multiple functional therapeutic effects of TnP: A small stable synthetic peptide derived from fish venom in a mouse model of multiple sclerosis.

Komegae EN, Souza TA, Grund LZ, Lima C, Lopes-Ferreira M
PloS one 2017;12(2):e0171796

---

DNA-binding of the Tet-transactivator curtails antigen-induced lymphocyte activation in mice.

Ottina E, Peperzak V, Schoeler K, Carrington E, Sgonc R, Pellegrini M, Preston S, Herold MJ, Strasser A, Villunger A
Nature communications 2017 Oct 18;8(1):1028

---

EZH2 enables germinal centre formation through epigenetic silencing of CDKN1A and an Rb-E2F1 feedback loop.

Béguelin W, Rivas MA, Calvo Fernández MT, Teater M, Purwada A, Redmond D, Shen H, Challman MF, Elemento O, Singh A, Melnick AM
Nature communications 2017 Oct 12;8(1):877

---

Differential cytokine contributions of perivascular haematopoietic stem cell niches.

Asada N, Kunisaki Y, Pierce H, Wang Z, Fernandez NF, Birbrair A, Ma'ayan A, Frenette PS
Nature cell biology 2017 Mar;19(3):214-223

---

Irgm1 coordinately regulates autoimmunity and host defense at select mucosal surfaces.

Azzam KM, Madenspacher JH, Cain DW, Lai L, Gowdy KM, Rai P, Janardhan K, Clayton N, Cunningham W, Jensen H, Patel PS, Kearney JF, Taylor GA, Fessler MB
JCI insight 2017 Aug 17;2(16)

---

Epithelial Fli1 deficiency drives systemic autoimmunity and fibrosis: Possible roles in scleroderma.

Takahashi T, Asano Y, Sugawara K, Yamashita T, Nakamura K, Saigusa R, Ichimura Y, Toyama T, Taniguchi T, Akamata K, Noda S, Yoshizaki A, Tsuruta D, Trojanowska M, Sato S
The Journal of experimental medicine 2017 Apr 3;214(4):1129-1151

---

Combined antiangiogenic and anti-PD-L1 therapy stimulates tumor immunity through HEV formation.

Allen E, Jabouille A, Rivera LB, Lodewijckx I, Missiaen R, Steri V, Feyen K, Tawney J, Hanahan D, Michael IP, Bergers G
Science translational medicine 2017 Apr 12;9(385)

---

Dendritic Cells Are Dispensable for T Cell Priming and Control of Acute Lymphocytic Choriomeningitis Virus Infection.

Hilpert C, Sitte S, Matthies A, Voehringer D
Journal of immunology (Baltimore, Md. : 1950) 2016 Oct 1;197(7):2780-6

---

Bmp signaling in colonic mesenchyme regulates stromal microenvironment and protects from polyposis initiation.

Allaire JM, Roy SA, Ouellet C, Lemieux É, Jones C, Paquet M, Boudreau F, Perreault N
International journal of cancer 2016 Jun 1;138(11):2700-12

---

Antigen presenting cell abnormalities in the Cln3(-/-) mouse model of juvenile neuronal ceroid lipofuscinosis.

Hersrud SL, Kovács AD, Pearce DA
Biochimica et biophysica acta 2016 Jul;1862(7):1324-36

---

Postnatal lethality and abnormal development of foregut and spleen in Ndrg4 mutant mice.

Qu X, Li J, Baldwin HS
Biochemical and biophysical research communications 2016 Feb 12;470(3):613-619

---

Murine iPSC-Derived Macrophages as a Tool for Disease Modeling of Hereditary Pulmonary Alveolar Proteinosis due to Csf2rb Deficiency.

Mucci A, Kunkiel J, Suzuki T, Brennig S, Glage S, Kühnel MP, Ackermann M, Happle C, Kuhn A, Schambach A, Trapnell BC, Hansen G, Moritz T, Lachmann N
Stem cell reports 2016 Aug 9;7(2):292-305

---

Resident T Cells Are Unable To Control Herpes Simplex Virus-1 Activity in the Brain Ependymal Region during Latency.

Menendez CM, Jinkins JK, Carr DJ
Journal of immunology (Baltimore, Md. : 1950) 2016 Aug 15;197(4):1262-75

---

The immunosenescence-related gene Zizimin2 is associated with early bone marrow B cell development and marginal zone B cell formation.

Matsuda T, Yanase S, Takaoka A, Maruyama M
Immunity & ageing : I & A 2015;12:1

---

Mitochondrial ATP transporter Ant2 depletion impairs erythropoiesis and B lymphopoiesis.

Cho J, Seo J, Lim CH, Yang L, Shiratsuchi T, Lee MH, Chowdhury RR, Kasahara H, Kim JS, Oh SP, Lee YJ, Terada N
Cell death and differentiation 2015 Sep;22(9):1437-50

---

Endothelial Gata5 transcription factor regulates blood pressure.

Messaoudi S, He Y, Gutsol A, Wight A, Hébert RL, Vilmundarson RO, Makrigiannis AP, Chalmers J, Hamet P, Tremblay J, McPherson R, Stewart AFR, Touyz RM, Nemer M
Nature communications 2015 Nov 30;6:8835

---

Limited miR-17-92 overexpression drives hematologic malignancies.

Danielson LS, Reavie L, Coussens M, Davalos V, Castillo-Martin M, Guijarro MV, Coffre M, Cordon-Cardo C, Aifantis I, Ibrahim S, Liu C, Koralov SB, Hernando E
Leukemia research 2015 Mar;39(3):335-41

---

Disruption of p21-activated kinase 1 gene diminishes atherosclerosis in apolipoprotein E-deficient mice.

Singh NK, Kotla S, Dyukova E, Traylor JG Jr, Orr AW, Chernoff J, Marion TN, Rao GN
Nature communications 2015 Jun 24;6:7450

---

Comparison of tamoxifen and letrozole response in mammary preneoplasia of ER and aromatase overexpressing mice defines an immune-associated gene signature linked to tamoxifen resistance.

Dabydeen SA, Kang K, Díaz-Cruz ES, Alamri A, Axelrod ML, Bouker KB, Al-Kharboosh R, Clarke R, Hennighausen L, Furth PA
Carcinogenesis 2015 Jan;36(1):122-32

---

Aging-associated inflammation promotes selection for adaptive oncogenic events in B cell progenitors.

Henry CJ, Casás-Selves M, Kim J, Zaberezhnyy V, Aghili L, Daniel AE, Jimenez L, Azam T, McNamee EN, Clambey ET, Klawitter J, Serkova NJ, Tan AC, Dinarello CA, DeGregori J
The Journal of clinical investigation 2015 Dec;125(12):4666-80

---

MHC-mismatched mixed chimerism augments thymic regulatory T-cell production and prevents relapse of EAE in mice.

Wu L, Li N, Zhang M, Xue SL, Cassady K, Lin Q, Riggs AD, Zeng D
Proceedings of the National Academy of Sciences of the United States of America 2015 Dec 29;112(52):15994-9

---

B-cell-intrinsic hepatitis C virus expression leads to B-cell-lymphomagenesis and induction of NF-κB signalling.

Kasama Y, Mizukami T, Kusunoki H, Peveling-Oberhag J, Nishito Y, Ozawa M, Kohara M, Mizuochi T, Tsukiyama-Kohara K
PloS one 2014;9(3):e91373

---

Transcription elongation factor ELL2 drives Ig secretory-specific mRNA production and the unfolded protein response.

Park KS, Bayles I, Szlachta-McGinn A, Paul J, Boiko J, Santos P, Liu J, Wang Z, Borghesi L, Milcarek C
Journal of immunology (Baltimore, Md. : 1950) 2014 Nov 1;193(9):4663-74

---

Rhof promotes murine marginal zone B cell development.

Kishimoto M, Matsuda T, Yanase S, Katsumi A, Suzuki N, Ikejiri M, Takagi A, Ikawa M, Kojima T, Kunishima S, Kiyoi H, Naoe T, Matsushita T, Maruyama M
Nagoya journal of medical science 2014 Aug;76(3-4):293-305

---

IκBε is a key regulator of B cell expansion by providing negative feedback on cRel and RelA in a stimulus-specific manner.

Alves BN, Tsui R, Almaden J, Shokhirev MN, Davis-Turak J, Fujimoto J, Birnbaum H, Ponomarenko J, Hoffmann A
Journal of immunology (Baltimore, Md. : 1950) 2014 Apr 1;192(7):3121-32

---

TDP2-dependent non-homologous end-joining protects against topoisomerase II-induced DNA breaks and genome instability in cells and in vivo.

Gómez-Herreros F, Romero-Granados R, Zeng Z, Alvarez-Quilón A, Quintero C, Ju L, Umans L, Vermeire L, Huylebroeck D, Caldecott KW, Cortés-Ledesma F
PLoS genetics 2013;9(3):e1003226

---

A component of the mir-17-92 polycistronic oncomir promotes oncogene-dependent apoptosis.

Olive V, Sabio E, Bennett MJ, De Jong CS, Biton A, McGann JC, Greaney SK, Sodir NM, Zhou AY, Balakrishnan A, Foth M, Luftig MA, Goga A, Speed TP, Xuan Z, Evan GI, Wan Y, Minella AC, He L
eLife 2013 Oct 15;2:e00822

---

Neuronal and nonneuronal cholinergic structures in the mouse gastrointestinal tract and spleen.

Gautron L, Rutkowski JM, Burton MD, Wei W, Wan Y, Elmquist JK
The Journal of comparative neurology 2013 Nov;521(16):3741-67

---

Natural IgG antibodies provide innate protection against ficolin-opsonized bacteria.

Panda S, Zhang J, Tan NS, Ho B, Ding JL
The EMBO journal 2013 Nov 13;32(22):2905-19

---

Reciprocal effects of rab7 deletion in activated and neglected T cells.

Roy SG, Stevens MW, So L, Edinger AL
Autophagy 2013 Jul;9(7):1009-23

---

Cbx4 regulates the proliferation of thymic epithelial cells and thymus function.

Liu B, Liu YF, Du YR, Mardaryev AN, Yang W, Chen H, Xu ZM, Xu CQ, Zhang XR, Botchkarev VA, Zhang Y, Xu GL
Development (Cambridge, England) 2013 Feb;140(4):780-8

---

Activation-induced cytidine deaminase-initiated off-target DNA breaks are detected and resolved during S phase.

Hasham MG, Snow KJ, Donghia NM, Branca JA, Lessard MD, Stavnezer J, Shopland LS, Mills KD
Journal of immunology (Baltimore, Md. : 1950) 2012 Sep 1;189(5):2374-82

---

Critical role of B cell lymphoma 10 in BAFF-regulated NF-κB activation and survival of anergic B cells.

Yu M, Chen Y, He Y, Podd A, Fu G, Wright JA, Kleiman E, Khan WN, Wen R, Wang D
Journal of immunology (Baltimore, Md. : 1950) 2012 Dec 1;189(11):5185-93

---

PAD4-mediated neutrophil extracellular trap formation is not required for immunity against influenza infection.

Hemmers S, Teijaro JR, Arandjelovic S, Mowen KA
PloS one 2011;6(7):e22043

---

Nuclear export of the NF-κB inhibitor IκBα is required for proper B cell and secondary lymphoid tissue formation.

Wuerzberger-Davis SM, Chen Y, Yang DT, Kearns JD, Bates PW, Lynch C, Ladell NC, Yu M, Podd A, Zeng H, Huang TT, Wen R, Hoffmann A, Wang D, Miyamoto S
Immunity 2011 Feb 25;34(2):188-200

---

A critical role of TAK1 in B-cell receptor-mediated nuclear factor kappaB activation.

Schuman J, Chen Y, Podd A, Yu M, Liu HH, Wen R, Chen ZJ, Wang D
Blood 2009 May 7;113(19):4566-74

---

p27 deficiency cooperates with Bcl-2 but not Bax to promote T-cell lymphoma.

Cheng N, van de Wetering CI, Knudson CM
PloS one 2008 Apr 2;3(4):e1911

---

Phospholipase Cgamma2 contributes to light-chain gene activation and receptor editing.

Bai L, Chen Y, He Y, Dai X, Lin X, Wen R, Wang D
Molecular and cellular biology 2007 Sep;27(17):5957-67

---

Essential role of phospholipase C gamma 2 in early B-cell development and Myc-mediated lymphomagenesis.

Wen R, Chen Y, Bai L, Fu G, Schuman J, Dai X, Zeng H, Yang C, Stephan RP, Cleveland JL, Wang D
Molecular and cellular biology 2006 Dec;26(24):9364-76

---

A role for brain-derived neurotrophic factor in B cell development.

Schuhmann B, Dietrich A, Sel S, Hahn C, Klingenspor M, Lommatzsch M, Gudermann T, Braun A, Renz H, Nockher WA
Journal of neuroimmunology 2005 Jun;163(1-2):15-23

---

Impaired immune responses and B-cell proliferation in mice lacking the Id3 gene.

Pan L, Sato S, Frederick JP, Sun XH, Zhuang Y
Molecular and cellular biology 1999 Sep;19(9):5969-80

---

Identification of monoclonal antibodies for immunohistochemical staining of feline B lymphocytes in frozen and formalin-fixed paraffin-embedded tissues.

Monteith CE, Chelack BJ, Davis WC, Haines DM
Canadian journal of veterinary research = Revue canadienne de recherche veterinaire 1996 Jul;60(3):193-8

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Staining of C57Bl/6 splenocytes with Anti-Mouse CD3e FITC (Product # 11-0031-82) and 0.125 µg of Rat IgG2a K Isotype Control eFluor® 660 (Product # 50-4321-82) (left) or 0.125 µg of Anti-Human/Mouse CD45R (B220) eFluor® 660 (right). Total viable cells were used for analysis.