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Cd45.1 apc

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CD45.1-APC is a fluorescently-labeled antibody that binds to the CD45.1 antigen. It is commonly used in flow cytometry applications to identify and quantify cell populations expressing the CD45.1 marker.

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Lab products found in correlation

Cd45.2 pe, by BD (3 mentions) Cd45.2 fitc, by BD (3 mentions) Cd3 fitc, by BD (3 mentions) Cd4 pe cy7, by BD (2 mentions) Cd11b pe cy7, by BD (2 mentions) B220 apc cy7, by BD (2 mentions) Lsfr fortessa, by FlowJo (2 mentions) Foxp3 fix perm kit, by Thermo Fisher Scientific (1 mentions) Cd138 pe, by BD (1 mentions) Diva software, by BD (1 mentions) Lsr 2, by BD (1 mentions) Irf4 pe, by Thermo Fisher Scientific (1 mentions) Irf8 percp efluor710, by Thermo Fisher Scientific (1 mentions) Cfse dilution, by Thermo Fisher Scientific (1 mentions) Facscalibur, by BD (1 mentions) Cd3 apc, by BD (1 mentions) Cd71 pe, by BD (1 mentions) Ter119 apc, by BD (1 mentions) C kit apc, by BD (1 mentions) B220 pe, by BD (1 mentions)

Close spelling variants of this product

CD45-APC (126 citations) Anti-CD45.1-APC (2 citations)

9 protocols using cd45.1 apc

1

Comprehensive B Cell Immunophenotyping

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B cell subsets were identified with CD22.2-FITC, CD138-PE, CD45.2-PE, CD45.1-APC (BD Biosciences), CD45R/B220-eFluor450, CD44-eFluor780 (eBioscience). RF B cells were detected with biotinylated-4G7 in combination with streptavidin-PerCP-Cy5.5. IRF-4 was detected using an IRF-4 antibody (clone M-17, Santa Cruz) and anti-goat IgG Alexa Fluor 647 (Jackson ImmunoResearch). IRF4-PE and IRF8-PercP-efluor710 (eBioscience) were used to co-stain IRF8 and IRF4. B cell proliferation was assessed by CFSE dilution (Life Technologies) (15 ) or VPD450 dilution (BD). Dead cells were distinguished with TO-PRO-3 (Life Technologies). To analyze TLR7 expression levels, unstimulated purified B cells, or B cells stimulated for 24 hr, were fixed and permeabilized using the Foxp3 Fix/Perm kit (ebioscience). TLR7 protein was detected using a biotinylated mouse TLR7 specific mAb, A94 (25 (link)), in combination with SA-PE. Flow cytometric analysis was carried out using a BD LSR II with Diva Software (BD) and analysis was conducted with FlowJo software (Tree Star).
Nündel K., Green N.M., Shaffer A.L., Moody K.S., Busto P., Eilat D., Miyake K., Oropallo M.A., Cancro M.P, & Marshak-Rothstein A. (2015). Cell intrinsic expression of TLR9 in autoreactive B cells constrains BCR/TLR7-dependent responses. Journal of immunology (Baltimore, Md. : 1950), 194(6), 2504-2512.
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2

Multiparametric Flow Cytometry Analysis of Hematopoietic Cells

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BM cells or splenocytes were harvested and subjected to red blood cell lysis. Fresh or frozen cells were stained with the following Abs: CD45.2-FITC and CD45.1-APC, Mac1-PE, Gr1-APC, c-Kit–APC, CD71-PE, Ter119-APC, B220-PE, and CD3-APC (BD) and analyzed on the BD FACSCalibur instrument. Staining for multiparameter flow cytometry was performed after a c-kit enrichment using 10 µl MACS beads (CD117) per mouse and then run on an AutoMACS (Miltenyi Biotec) according to the manufacturer’s instructions. The cells were then stained with the following cocktail: (Lineage; CD3, CD4, CD8, Gr1, B220, CD19, and TER119 all conjugated with PeCy5), Sca-Pac Blue, CD34-FITC or CD45.2-FITC, SLAM-APC, CD48-PE, c-KIT–Alexa Fluor 780, and FcgRIIb-PeCy7 (Fig. 2 d); HPCs (Linloc-Kit+Sca1−), GMPs (LK, FcγRIIbhiCD34+), CMPs (LK, FcγRIIbmid CD34+), MEPs (LK, FcγRIIbloCD34−), B cells (B220+), and T cells (CD3+) from the spleen were also sorted. For analysis of LMPPs and CLPs, the following cocktail was used: Lineage marker mix–PeCy5, Sca-Pecy7 IL-7Ra–Pac Blue, Flk2-PE, CD34-FITC, and Kit-APC (Martin et al., 2003 (link); Adolfsson et al., 2005 (link); Karsunky et al., 2008 (link)).
Park S.M., Deering R.P., Lu Y., Tivnan P., Lianoglou S., Al-Shahrour F., Ebert B.L., Hacohen N., Leslie C., Daley G.Q., Lengner C.J, & Kharas M.G. (2014). Musashi-2 controls cell fate, lineage bias, and TGF-β signaling in HSCs. The Journal of Experimental Medicine, 211(1), 71-87.
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3

Immunophenotyping of Mouse Bone Marrow

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Bone-marrow cells were flushed from the long bones (tibias and femurs) and stained in PBS (Corning Cellgro) supplemented with 2% of inactivated fetal bovine serum (Gemini BioProducts) with the following antibodies from BD Biosciences: B220 FITC, CD11b FITC, PE, APC or APCcy7, CD11CFITC, PE, APC or APC-cy7, CD4FITC, PE or APC, CD8 FITC, PE or APC, NK1.1 FITC, Ter119 FITC, CD3 FITC (when gates on lineage negative bone-marrow cells were used, FITC-conjugated antibodies were used), c-Kit PE or APC, Flk2 PE, CD45.2 biotin or FITC, CD45.1 APC, IL7R Alexa 647, CD34 APC, CD150 PE. From eBiosciences: Sca-1 PEcy7, AA4.1 bio or PEcy-7, streptavidin APC. From Invitrogen: streptavidin Pacific blue. DAPI was used to exclude dead cells. Flow cytometry was performed on a fluorescence-activated cell sorting (FACS) Calibur, LSRII or LSR Fortessa (BD Biosciences).
Santos M.A., Faryabi R.B., Ergen A.V., Day A.M., Malhowski A., Canela A., Onozawa M., Lee J.E., Callen E., Gutierrez-Martinez P., Chen H.T., Wong N., Finkel N., Deshpande A., Sharrow S., Rossi D.J., Ito K., Ge K., Aplan P.D., Armstrong S.A, & Nussenzweig A. (2014). DNA-damage-induced differentiation of leukaemic cells as an anti-cancer barrier. Nature, 514(7520), 107-111.
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4

Murine Immune Cell Phenotyping

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Murine GM-CSF was from Peprotech (Neuilly-sur-Seine, France). IL-6 and LPS were from Sigma-Aldrich (Saint-Quentin Fallavier, France). CFDA-SE (CFSE) was from Molecular Probes (Montluçon, France). OVA (SIINFEKL) and Smcy (KCSRNRQYL) peptides were from PolyPeptide (Strasbourg, France). Anti mouse CD11b biotin (M1/70) (used with streptavidin APC or streptavidin APC-Cy7), CD11b APC-Cy7 (M1/70), CD11c PE-Cy7 (HL3), I-Ab FITC (AF6-120.1), Gr1 PE (Ly6C/G, RB6-8C5), CD45.1 APC (A20), CD45.2 APC-Cy7 (104), CD45.2 PerCP-Cy5.5 (104), CD19 APC (1D3), NK1.1 PE (PK136), CD3ε PerCP-Cy5.5 (145-2C11), CD3ε Pacific Blue (500A2), CD3ε FITC (145-2C11), CD4 PE-Cy7 (RM4-5), CD8α Pacific blue (53-6.7), CD8α APC-Cy7 (53-6.7), CD8α PerCP-Cy5.5 (53-6.7), FoxP3 Alexa Fluor647 (MF23), CD25 PE (704), CD69 FITC (H1.2F3), and CD86 FITC (B7.2, GL1) were from BD PharMingen (Le Pont de Claix, France). Male antigen UTY-specific CD8+ T cells were detected using a PE labelled Pro5 MHC Pentamer (H-2Db, WMHHNMDLI) (ProImmune Limited, Oxford, UK).
Drujont L., Carretero-Iglesia L., Bouchet-Delbos L., Beriou G., Merieau E., Hill M., Delneste Y., Cuturi M.C, & Louvet C. (2014). Evaluation of the Therapeutic Potential of Bone Marrow-Derived Myeloid Suppressor Cell (MDSC) Adoptive Transfer in Mouse Models of Autoimmunity and Allograft Rejection. PLoS ONE, 9(6), e100013.
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5

Evaluating Engraftment of Gene-Modified Cells

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At 7-and 17-week post-transplant, peripheral blood was collected from study animals and analyzed by flow cytometry (FACS) and qPCR to evaluate engraftment of gene-modified cells. Blood was collected from the lateral tail vein in an EDTA-containing tube. RBC Lysis was carried out using an ammonium chloride-based method (Pharmlyse, BO), and the cells were divided for qPCR or FACS analysis. Cells for FACS were stained with antibodies against CD45.1 and CD45.2 to distinguish between host and donor cells and against CD 11b, CD3 and B220 for immunophenotypic characterization (Antibodies used: CD45.1-APC, CD45.2-PE, CD 11b-PECy7, CD3-FITC and B220-APC-Cy7; all from BD Biosciences, San Jose, CA; cat.no. 561873, 560695, 552850, 555274, and 552094, respectively).
Shadid M., Shrestha A, & Malik P. (2024). Preclinical safety assessment of modified gamma globin lentiviral vector-mediated autologous hematopoietic stem cell gene therapy for hemoglobinopathies. PloS one, 19(7).
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6

Comprehensive Immunophenotyping of Murine Cells

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Single cell suspensions from thymus (if large enough), spleen, and bone marrow (from sacrificed animals) were prepared and cells were counted using a hemacytometer or automated cell counter. A subset of cells (approximately 1 x 10 6 ) was used for flow cytometric analyses. In addition, peripheral blood samples were also analyzed by Flow Cytometry. Surface markers used for immunophenotyping included CD45.1, CD45.2, CD 11b, CD8, CD3, CD4, B220 and Gr-I (Antibodies used: CD45.1-APC, CD45.2-PE, CD 11b-PECy7, CD3-FITC and B220-APC-Cy7, CD4-PE-Cy7, CD8a-APC-Cy7, and Gr-1-APC-cy7all from BD Biosciences, San Jose, CA; cat.no. 561873, 560695, 552850, 555274, 552094, 563933, 557654, and 560600, respectively).
Shadid M., Shrestha A, & Malik P. (2024). Preclinical safety assessment of modified gamma globin lentiviral vector-mediated autologous hematopoietic stem cell gene therapy for hemoglobinopathies. PloS one, 19(7).
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7

Phenotyping Murine and Human Leukemic Cells

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Murine leukemic cells were analyzed by flow cytometry using the following antibodies: CD45.1-APC (BD PharMingen, USA), Lineage cocktail (StemCell Technologies Inc, Canada), Sca-1-PECy7 and c-kit-PE (BD PharMingen). For human leukemic cells, CD45-APC, CD34-BV421, CD90-FITC (BD PharMingen) antibodies were used. For MSC, anti-CD106 (VCAM-1)-biotin, CD51-PE (eBioscience, CA, USA.), CD140a (PDGFRa)-APC, Sca- 1-PE-Cy7 (BD PharMingen) were used.
Lee H.R., Lee G.Y., Kim E.W., Kim H.J., Lee M., Humphries R.K, & Oh I.H. (2021). Reversible switching of leukemic cells to a drugresistant, stem-like subset via IL-4-mediated cross-talk with mesenchymal stroma. Haematologica, 107(2), 381-392.
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8

Competitive Hematopoietic Stem Cell Transplantation

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C57BL/6 mice (n=4 per group) were irradiated with a lethal dose of 2 × 500 rad. They were then injected into the tail vein with a combination of 250 000 lineage negative bone marrow cells (lin−) from C57BL/6.SJL (Sf3b1+/+, CD45.1) mice and 250 000 lin− cells from Sf3b1K700E/+ (CD45.2) mice plus 250 000 whole bone marrow (BM) rescue cells from C57BL/6/6.SJL (CD45.1/CD45.2) mice. Control animals were injected with 1  ×  106 whole BM cells from C57BL/6/6.SJL (CD45.1/CD45.2). Blood counts and FACS analysis were performed after 1, 2 and 4 months. Samples were stained with CD45.1-APC (Becton Dickinson, Oxford, UK) and CD45.2-FITC (Becton Dickinson) antibodies run on Becton Dickinson LSFR Fortessa and analyzed with Flowjo 7.6.5 (FlowJo, LLC, San Diego, CA, USA).
Mupo A., Seiler M., Sathiaseelan V., Pance A., Yang Y., Agrawal A.A., Iorio F., Bautista R., Pacharne S., Tzelepis K., Manes N., Wright P., Papaemmanuil E., Kent D.G., Campbell P.C., Buonamici S., Bolli N, & Vassiliou G.S. (2016). Hemopoietic-specific Sf3b1-K700E knock-in mice display the splicing defect seen in human MDS but develop anemia without ring sideroblasts. Leukemia, 31(3), 720-727.
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9

Competitive Hematopoietic Engraftment Assay

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C57/BL6 mice were irradiated with a lethal dose of 2x500rad. They were then injected into the tail vein with a combination of 250.000cells of BM-ve cells from C56/Bl6.SJL (Sf3b1+/+, CD45.1) mice and 250.00 BM-ve cells from Sf3b1K700E/+ (CD45.2) mice plus 250.000 whole BM rescue cells from C57/Bl6 (CD45.1/CD45.2) mice. Control animals were injected with 1x106 whole BM cells from C57/Bl6 (CD45.1/CD45.2). Blood counts and FACS analysis were performed monthly. Samples were stained with CD45.1-APC (Becton Dickinson, UK), CD45.2-FITC (Becton Dickinson) antibodies runned on Becton Dickinson LSFR Fortessa and analyzed with Flowjo 7.6.5 (FlowJo, LLC USA).
Mupo A., Seiler M., Sathiaseelan V., Pance A., Yang Y., Agrawal A.A., Iorio F., Bautista R., Pacharne S., Tzelepis K., Manes N., Wright P., Papaemmanuil E., Kent D.G., Campbell P.C., Buonamici S., Bolli N, & Vassiliou G.S. (2016). Hemopoietic-specific Sf3b1-K700E knock-in mice display the splicing defect seen in human MDS but develop anemia without ring sideroblasts. Leukemia, 31(3), 720-727.
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