Antibody data
- Antibody Data
- Antigen structure
- References [9]
- Comments [0]
- Validations
- Flow cytometry [1]
- Other assay [8]
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- Product number
- 35-0209-42 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- CD20 Monoclonal Antibody (2H7), PE-Cyanine5.5, eBioscience™
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- Description: The 2H7 monoclonal antibody reacts with human CD20, a 33-36 kDa transmembrane protein. CD20 is expressed by developing B cells as well as mature B cells but not plasma cells. CD20 has been detected at low levels on a small subset of mature T cells. It is suggested that CD20 plays a role in B-cell activation. Applications Reported: This 2H7 antibody has been reported for use in flow cytometric analysis. Applications Tested: This 2H7 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 µL (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. Light sensitivity: This tandem dye is sensitive photo-induced oxidation. Please protect this vial and stained samples from light. Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-822-49) (100 µL cell sample + 100 µL IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333-54) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically. Excitation: 488-561 nm; Emission: 695 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser.
- Reactivity
- Human
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- 2H7
- Vial size
- 100 Tests
- Concentration
- 5 µL/Test
- Storage
- 4° C, store in dark, DO NOT FREEZE!
Submitted references TRPA1 triggers hyperalgesia and inflammation after tooth bleaching.
Genetic modification of primary human B cells to model high-grade lymphoma.
Relative safety of various spermatogenic stem cell purification methods for application in spermatogenic stem cell transplantation.
Evidence for persistence of the SHIV reservoir early after MHC haploidentical hematopoietic stem cell transplantation.
Tumor-associated B-cells induce tumor heterogeneity and therapy resistance.
Phase I study of a bispecific ligand-directed toxin targeting CD22 and CD19 (DT2219) for refractory B-cell malignancies.
Immunologic human renal allograft injury associates with an altered IL-10/TNF-α expression ratio in regulatory B cells.
Transcriptional profiling of mRNAs and microRNAs in human bone marrow precursor B cells identifies subset- and age-specific variations.
An efficient low cost method for gene transfer to T lymphocytes.
Chen C, Huang X, Zhu W, Ding C, Huang P, Li R
Scientific reports 2021 Aug 31;11(1):17418
Scientific reports 2021 Aug 31;11(1):17418
Genetic modification of primary human B cells to model high-grade lymphoma.
Caeser R, Di Re M, Krupka JA, Gao J, Lara-Chica M, Dias JML, Cooke SL, Fenner R, Usheva Z, Runge HFP, Beer PA, Eldaly H, Pak HK, Park CS, Vassiliou GS, Huntly BJP, Mupo A, Bashford-Rogers RJM, Hodson DJ
Nature communications 2019 Oct 4;10(1):4543
Nature communications 2019 Oct 4;10(1):4543
Relative safety of various spermatogenic stem cell purification methods for application in spermatogenic stem cell transplantation.
Tian J, Ma K, Pei CB, Zhang SH, Li X, Zhou Y, Yan B, Wang HY, Ma LH
Stem cell research & therapy 2019 Dec 16;10(1):382
Stem cell research & therapy 2019 Dec 16;10(1):382
Evidence for persistence of the SHIV reservoir early after MHC haploidentical hematopoietic stem cell transplantation.
Colonna L, Peterson CW, Schell JB, Carlson JM, Tkachev V, Brown M, Yu A, Reddy S, Obenza WM, Nelson V, Polacino PS, Mack H, Hu SL, Zeleski K, Hoffman M, Olvera J, Furlan SN, Zheng H, Taraseviciute A, Hunt DJ, Betz K, Lane JF, Vogel K, Hotchkiss CE, Moats C, Baldessari A, Murnane RD, English C, Astley CA, Wangari S, Agricola B, Ahrens J, Iwayama N, May A, Stensland L, Huang MW, Jerome KR, Kiem HP, Kean LS
Nature communications 2018 Oct 25;9(1):4438
Nature communications 2018 Oct 25;9(1):4438
Tumor-associated B-cells induce tumor heterogeneity and therapy resistance.
Somasundaram R, Zhang G, Fukunaga-Kalabis M, Perego M, Krepler C, Xu X, Wagner C, Hristova D, Zhang J, Tian T, Wei Z, Liu Q, Garg K, Griss J, Hards R, Maurer M, Hafner C, Mayerhöfer M, Karanikas G, Jalili A, Bauer-Pohl V, Weihsengruber F, Rappersberger K, Koller J, Lang R, Hudgens C, Chen G, Tetzlaff M, Wu L, Frederick DT, Scolyer RA, Long GV, Damle M, Ellingsworth C, Grinman L, Choi H, Gavin BJ, Dunagin M, Raj A, Scholler N, Gross L, Beqiri M, Bennett K, Watson I, Schaider H, Davies MA, Wargo J, Czerniecki BJ, Schuchter L, Herlyn D, Flaherty K, Herlyn M, Wagner SN
Nature communications 2017 Sep 19;8(1):607
Nature communications 2017 Sep 19;8(1):607
Phase I study of a bispecific ligand-directed toxin targeting CD22 and CD19 (DT2219) for refractory B-cell malignancies.
Bachanova V, Frankel AE, Cao Q, Lewis D, Grzywacz B, Verneris MR, Ustun C, Lazaryan A, McClune B, Warlick ED, Kantarjian H, Weisdorf DJ, Miller JS, Vallera DA
Clinical cancer research : an official journal of the American Association for Cancer Research 2015 Mar 15;21(6):1267-72
Clinical cancer research : an official journal of the American Association for Cancer Research 2015 Mar 15;21(6):1267-72
Immunologic human renal allograft injury associates with an altered IL-10/TNF-α expression ratio in regulatory B cells.
Cherukuri A, Rothstein DM, Clark B, Carter CR, Davison A, Hernandez-Fuentes M, Hewitt E, Salama AD, Baker RJ
Journal of the American Society of Nephrology : JASN 2014 Jul;25(7):1575-85
Journal of the American Society of Nephrology : JASN 2014 Jul;25(7):1575-85
Transcriptional profiling of mRNAs and microRNAs in human bone marrow precursor B cells identifies subset- and age-specific variations.
Jensen K, Brusletto BS, Aass HC, Olstad OK, Kierulf P, Gautvik KM
PloS one 2013;8(7):e70721
PloS one 2013;8(7):e70721
An efficient low cost method for gene transfer to T lymphocytes.
Chicaybam L, Sodre AL, Curzio BA, Bonamino MH
PloS one 2013;8(3):e60298
PloS one 2013;8(3):e60298
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Supportive validation
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- Staining of normal human peripheral blood cells with Anti-Human CD3 FITC (Product # 11-0038-42) and Mouse IgG2b K Isotype Control PE-Cyanine5.5 (Product # 35-4732-80) (left) or Anti-Human CD20 PE-Cyanine5.5 (right). Cells in the lymphocyte gate were used for analysis.
Supportive validation
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- Figure 1 Cell sorting of precursor B cells subsets from CD10 positively selected cells. Immunomagnetic selection and subsequent FACS were used to isolate the five populations from pediatric and adult human BM. Shown are the FACS dot plots with sorting gates to obtain CD34 + CD19 - ProB cells, CD34 + CD19 + PreBI cells, CD34 - CD19 + CD20 dim PreBII large cells, CD34 - CD19 + CD20 - PreBII small cells, and CD34 - CD19 + CD20 high IgM + Immature B cells.
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- Fig. 4 IGF-1-dependent induction of cancer stem cell markers CD20, CD133, and CD271 (NGFR) on melanoma cells. a Melanoma cells (WM3749) co-cultured with TAB cells (days 3 and 9) show high expression of CD20 ( middle and right panels ) compared with the control culture ( left panel ) as determined by FACS analysis. Melanoma cells were co-stained with anti-CD146 (MCAM, PE-conjugated) and anti-CD20 (FITC-conjugated) antibodies to distinguish them from B cells, which are CD146-negative;percentages indicate co-expression of both markers on the malignant cells. b Melanoma cells (WM3749) co-cultured with TAB cells (day 6) show high expression of CD20, CD133 and CD271 ( left panel ) compared with minimal or low expression of those markers when tumor cells are co-cultured with NB cells ( right panel ). Co-culture of melanoma cells with TAB cells did not modulate the expression of CD144 (vascular-endothelial cadherin marker) that are normally expressed by aggressive melanomas (data not shown). Induction of CD20, CD133, and CD271 was blocked when anti-IGF-1 neutralizing antibody (10 mug/ml) was used in the co-culture ( middle panel ). Anti-IL-1, anti-PDGF or anti-VEGF antibodies had no effect on CD marker expression (data not shown). Percentages indicate co-expression of CD20, CD133, or CD271on CD146 + melanoma cells. Results are representative of two independent experiments. c Melanoma cells(WM3749, WM989 and 451Lu) cultured in the presence of recombinant IGF-1 (25 ng/ml) for 5 days sh
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- Fig. 2 Long-term expansion of human germinal center B cells ex vivo. a Primary human GC B cells were transduced with the indicated oncogenes and oncogene combinations and cultured separately for up to 120 days. Graph shows calculated theoretical absolute cell numbers (+-s.e.m., n = 3). Viable cells were assessed by trypan blue exclusion. Source data are provided as a Source Data file. b Primary human GC B cells were transduced with different oncogenes and oncogene combinations and monitored by flow cytometry. Graph shows the change in cell viability assessed by scatter characteristic by flow cytometry ( +- s.e.m., n = 3). Source data are provided as a Source Data file. c Primary human GC B cells were transduced with BCL2 in combination with other transcription factors in a pooled, competitive culture. Graph shows relative abundance of transcription factors or their mutant versions over four different timepoints ( n = 3). d Primary human GC B cells were transduced with the oncogenic cocktail BCL2 and BCL6 and cultured to day 73. Representative flow cytometry analysis ( n = 3) for the expression of the GC B cell markers CD38, CD20, CD19, CD80, CD22, CD95, CXCR4, and CD86. Red histograms show GC B cells compared to primary human naive B cells (blue). e Heat map of gene expression of freshly isolated GC B cells ( n = 3), transduced GC B cells ( BCL2-BCL6, BCL2-MYC ) cultured ex vivo for 5 or 73 days ( n = 3), plasma cell line ( n = 1), naive B cells ( n = 1), and lymphoma cell li
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- Figure 1 Identification of dental pulp stem cells (DPSCs). Human DPSCs were positive for the cell surface antigens CD73, CD90, and CD105, as well as negative for CD14, CD20, CD34, and CD45 demonstrated by flow cytometry ( A ). DPSCs were cultured under osteogenic ( B , 14 days) or adipogenic ( C , 21 days) conditions, and showed mineralized nodules and lipid clusters as revealed by alizarin red and oil red staining, respectively. Scale bar = 400 ( B ) or 100 ( C ) mum.
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- Fig. 3 Allogeneic HCT in SHIV-C Infected, cART-treated RM. a Schematic of SHIV-C infection and allogeneic bone marrow transplantation strategy in SHIV-C infected, cART-treated RM. b Plasma Viral Loads (PVL) were measured longitudinally following transplant as specified in Fig. 1a . c White blood count (WBC) (x10 3 /uL). d Absolute Neutrophil Count (ANC) (x10 3 /uL). e Absolute Lymphocyte Count (ALC) (x10 3 /uL). f Platelet Count (PLT) (x10 3 /uL). g Hemoglobin (Hgb) concentration (g/dL). h Percent whole blood donor chimerism, measured by microsatellite analysis. i Percent donor myeloid (CD11b + CD3-) chimerism, measured flow cytometrically. j Percent donor CD4 + T cell (CD3+ CD4+ CD8- CD20- CD11b- lymphocytes) chimerism, measured flow cytometrically. k Percent donor CD8 + T cell (CD3+ CD8+ CD4- CD20- CD11b- lymphocytes) chimerism, measured by flow cytometry
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- Fig. 1 Marker verification and validation of independent markers in BALL cells by immunofluorescence and flow cytometry. The expression of CD20 ( a - c ) and CD38 ( d - f ), were positive on BALL cell membranes. CD90f ( g - i ) and CD49f ( j , k ) were not expressed in BALL cells. DAPI indicates the cell nucleus. This finding was confirmed via the conducted flow cytometry analysis. Scale bar = 100 mum