Igg1 apc

Manufactured by Miltenyi Biotec

Sourced in United States, Germany

IgG1-APC is a mouse monoclonal antibody conjugated to the fluorochrome allophycocyanin (APC). It is designed for flow cytometry applications to detect and analyze cells expressing IgG1.

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Lab products found in correlation

Igg1 pe, by Miltenyi Biotec (6 mentions) Igg2a percp, by Miltenyi Biotec (3 mentions) Cd45 percp, by Miltenyi Biotec (3 mentions) Igg1 fitc, by Miltenyi Biotec (2 mentions) Igg1 pe vio770, by Miltenyi Biotec (2 mentions) Cd73 fitc, by Thermo Fisher Scientific (2 mentions) Cd271 apc, by Miltenyi Biotec (2 mentions) Human cd105 pe, by Thermo Fisher Scientific (2 mentions) Cd44 fitc, by Miltenyi Biotec (2 mentions) α rabbit alexafluor 488, by Thermo Fisher Scientific (1 mentions) α mouse alexafluor488, by Thermo Fisher Scientific (1 mentions) α phospho histone h3, by Cell Signaling Technology (1 mentions) α mouse alexa fluor 568, by Thermo Fisher Scientific (1 mentions) α olig2, by Merck Group (1 mentions) Lsrii flow cytometer, by BD (1 mentions) Pharm lyse buffer, by BD (1 mentions) Igg1 pe, by BD (1 mentions) Facsdiva software, by BD (1 mentions) Anti cd24 apc, by Miltenyi Biotec (1 mentions) Anti cd44 pe, by Miltenyi Biotec (1 mentions)

Close spelling variants of this product

Mouse IgG1-APC (9 citations) APC-conjugated mouse IgG1 (2 citations) REA control human IgG1 APC-conjugated isotype antibody (2 citations)

10 protocols using igg1 apc

Immunophenotyping of Cell Populations

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The antibodies used in this study were α-CD44 (Dako, M708201), α-CD44-FITC (Miltenyi-Biotec, 130-098-210), α-CD133-APC (Miltenyi-Biotech, 130-098-829), α-phospho-histone H3 (Cell signaling, 9701), IgG1-FITC (Miltenyi-Biotec, 130-098-847), IgG1-APC (Miltenyi-Biotech, 130-098- 846), α-Olig2 (Millipore, ab9610), α-Mouse Alexa Fluor 488 (ThermoFisher Scientific, A11029), α-Mouse Alexa Fluor 568 (ThermoFisher Scientific, A11004), α-Rabbit Alexa Fluor 488 (ThermoFisher Scientific, A11008) and α-Rabbit Alexa Fluor 568 (ThermoFisher Scientific, A11011).

Brown D.V., Filiz G., Daniel P.M., Hollande F., Dworkin S., Amiridis S., Kountouri N., Ng W., Morokoff A.P, & Mantamadiotis T. (2017). Expression of CD133 and CD44 in glioblastoma stem cells correlates with cell proliferation, phenotype stability and intra-tumor heterogeneity. PLoS ONE, 12(2), e0172791.

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Quantification of Circulating Stem Cells

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The full population of PBNCs was obtained after the lysis of erythrocytes using BD Pharm Lyse Buffer (Pharmingen, BD Biosciences, San Diego, CA, USA). For flow cytometry, 1 million NCs were resuspended in 100 μL of phosphate-buffered saline (PBS). Immunofluorescence cell staining was performed using a phycoerythrin- (PE-) conjugated anti-CD309 (VEGFR-2/KDR) antibody (Pharmingen, BD Biosciences, San Jose, USA; clone 89106) and an allophycocyanin- (APC-) conjugated anti-CD133 antibody (Miltenyi Biotec, Auburn, CA, USA; clone AC133). Samples stained with appropriate isotype control antibodies, specifically IgG1-PE (Pharmingen, BD Biosciences, San Jose, USA; clone MOPC-21) and IgG1-APC (Miltenyi Biotec, Auburn, CA, USA), were prepared in parallel and served as negative controls. After 20 min incubation on ice, the cells were washed twice in PBS and resuspended in 1% paraformaldehyde. Cell fluorescence was measured, and data were analyzed using an LSRII flow cytometer (BD Biosciences, San Jose, CA, USA) with BD FACSDiva software. Typically, 2 × 10⁵ events were acquired to determine the proportions of examined subpopulations within the PBNCs. Using this approach, populations of circulating stem cells (CD133⁺) and early EPCs (CD133⁺/VEGFR2⁺) were analyzed.

Gołąb-Janowska M., Paczkowska E., Machaliński B., Kotlęga D., Meller A., Safranow K., Maj M, & Nowacki P. (2018). Effects of Angiotensin-Converting Enzyme Inhibition on Circulating Endothelial Progenitor Cells in Patients with Acute Ischemic Stroke. Stem Cells International, 2018, 2827580.

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Flow Cytometric Analysis of Cancer Stem Cell Markers

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To detect the expression of cancer stem cell markers, the following antibodies were used: anti-CD44-PE, anti-CD24-APC, anti-ESA-PE-Vio770, IgG1-PE, IgG1-APC, IgG1-PE-Vio770 (Miltenyi). Cells were harvested with trypsin-EDTA to produce a single cell suspension. The cells were pelleted by centrifugation, washed twice with PBS and incubated with the antibody for 10 min in the dark at 4℃. The respective isotype control antibodies were used at the same concentrations according to the manufacturer's instructions. After washing twice with PBS, samples were resuspended in 500 μl PBS and analyzed on a flow cytometer (FACSAriaII, USA). Cells were routinely sorted twice, and the cells were reanalyzed for the purity, which typically was >97%. Data were analyzed with BD FACS Diva software.

Wang F., Ma L., Zhang Z., Liu X., Gao H., Zhuang Y., Yang P., Kornmann M., Tian X, & Yang Y. (2016). Hedgehog Signaling Regulates Epithelial-Mesenchymal Transition in Pancreatic Cancer Stem-Like Cells. Journal of Cancer, 7(4), 408-417.

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Characterization of ATD-MSCs by Flow Cytometry

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Cell surface markers of ATD-MSCs were analysed by flow cytometry on fresh SVF and on SVF derived after thawing of adipose tissue and its collagenase treatment. ATD-MSCs were identified as CD105, CD44, CD73, and CD271 positive cells and negative for CD45 expression. Standard labelling protocol was performed with the following antibodies fluorochrome-conjugated and isotypic controls: human CD105 PE (Invitrogen), CD73 FITC (kindly provided by Professor Malavasi, University of Turin), CD44 FITC, CD45 PerCP, CD3 PerCP, CD271 APC, IgG1 PE, IgG1 APC, and IgG2a PerCP (Miltenyi Biotec), and IgG1 FITC-conjugated (Immunostep). About 10⁵ events/sample were used for capture with CellQuest software. All data were analysed with FlowJo software (Tree Star).

Roato I., Alotto D., Belisario D.C., Casarin S., Fumagalli M., Cambieri I., Piana R., Stella M., Ferracini R, & Castagnoli C. (2016). Adipose Derived-Mesenchymal Stem Cells Viability and Differentiating Features for Orthopaedic Reparative Applications: Banking of Adipose Tissue. Stem Cells International, 2016, 4968724.

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Multiparameter Flow Cytometry for Plasma Cell Phenotyping

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Flow cytometry was performed with 6-color direct immunofluorescence detection using a Cytoflex LX (Beckman Coulter, Brea, CA, USA). Staining was performed with the following antibodies: CD19 PE (LT19, Miltenyi 130113169), CD138 APC (44F9, Miltenyi 130117395), CD20 e450 (2H7, Thermo Fischer 48020942) (Thermo Fischer, Waltham, MA, USA), CD27 FITC (M-T271, BD 55540), and CD38 PECy7 (HB7, BD 335825). Isotype-matched antibodies were used as controls (IgG1—PE Miltenyi 130113200; IgG1—APC Miltenyi 130113196; IgG2b—eFlour450 Thermo Fisher 48473282; IgG1—FITC BD 348808; IgG1—PECy7 Miltenyi 130113196,). The zombie UV fixable viability kit (Biolegend 423108, San Diego, CA, USA) was used to exclude dead cells. FlowJo version 10 (BD Biosciences, Franklin Lakes, NJ, USA) and Prism 9 (GraphPad, San Diego, CA, USA) with Spearman correlation statistical analysis were applied for analysis.

Care M.A., Stephenson S., Owen R., Doody G.M, & Tooze R.M. (2023). Spontaneous EBV-Reactivation during B Cell Differentiation as a Model for Polymorphic EBV-Driven Lymphoproliferation. Cancers, 15(12), 3083.

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Mesenchymal Cell Surface Marker Analysis

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Mesenchymal cell surface markers were analysed by flow cytometry on fresh SVF and on cultured ASCs. A standard labelling protocol was performed with the following antibody fluorochrome-conjugated and isotypic controls: human CD105 PE (Invitrogen), CD73 FITC (kindly provided by Prof. Malavasi, University of Turin), CD44 FITC, CD45 PerCP, CD271 APC, IgG1 PE, IgG1 APC and IgG2a PerCP (Miltenyi Biotec), and IgG1 FITC conjugate (IMMUNOSTEP). About 10⁵ events/sample were used for capture with CellQuest software. All data were analysed with Flowlogic software (Miltenyi Biotec).

Roato I., Belisario D.C., Compagno M., Verderio L., Sighinolfi A., Mussano F., Genova T., Veneziano F., Pertici G., Perale G, & Ferracini R. (2018). Adipose-Derived Stromal Vascular Fraction/Xenohybrid Bone Scaffold: An Alternative Source for Bone Regeneration. Stem Cells International, 2018, 4126379.

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Stromal Vascular Fraction Analysis

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MFAT samples were treated with collagenase IV (Serva GmbH) to retrieve SVF and analyze its various cell components, in particular the ASCs, expressing CD105 + , CD73 + , CD90 + and CD44 + and negative for HLA-DR, CD34, CD31 and CD45; CD146 + pericytes, CD31 + endothelial cells, CD34 + hematopoietic cells. The standard labelling protocol was performed with the following fluorochrome-conjugated antibodies and isotypic controls: ASCs were marked with MSC phenotyping kit, human CD44 PE, CD45 PerCP, IgG1 PE, IgG1 FITC, IgG1 APC, and IgG2a PerCP (Miltenyi Biotec, Bergisch Gladbach, Germany), CD31PE, CD34FITC, CD146APC and CD90 PerCP (Biolegend, San Diego, CA, USA). About 10⁵ events/sample were used for capture with MACsQUANT10 and data were analyzed with MACs quantify software (Miltenyi Biotec, Bergisch Gladbach, Germany). The percentage of ASCs in the MFAT volume was determined, and then the number of ASCs for ml of MFAT was normalized on the absolute number of viable cells.

Ferracini R., Alessio-Mazzola M., Sonzogni B., Stambazzi C., Ursino C., Roato I., Mussano F., Bistolfi A., Furlan S., Godio L., Alotto D, & Formica M. (2022). Age and synovitis affect the results of the treatment of knee osteoarthritis with Microfragmented Autologous Fat Tissue. Knee Surgery, Sports Traumatology, Arthroscopy, 31(9), 3655-3664.

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Comprehensive Immune Profile Analysis

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Clinical parameters [complete blood count with differential, CD4 count, HCV and HIV viral load (VL), alanine aminotransferase (ALT), and aspartate aminotransferase (AST)] were assessed by Quest Diagnostics (NJ, USA).
Flow cytometry was performed on fresh blood as previously described ^{[19 (link), 20 ]}. Combinations of fluorochrome-conjugated monoclonal Abs targeted activation/exhaustion, costimulatory and apoptosis markers as shown in Supplemental Table 2. All Abs were from Becton Dickinson Biosciences (San Diego, CA, USA) except blood dendritic cells antigen (BDCA) 2-allophycocyanin (APC), BDCA4-APC and IgG1-APC (Miltenyi Biotec, San Diego, CA, USA). T cells were defined as CD3⁺CD8⁻, CD3⁺CD8⁺, dendritic cells (DC) ^{[21 (link)]} as BDCA2⁺BDCA4⁺ (plasmacytoid DC, PDC), and CD19⁻BDCA1⁺CD11c⁺ (myeloid DC, MDC)], monocytes as CD14⁺, and NK cells as CD3⁻CD56^dimCD16⁻, CD3⁻CD56^dimCD16⁺, CD3⁻CD56^bright, and CD3⁻CD56⁻CD16^{+ [22 (link)–25 (link)]}.

GIRON L.B., AZZONI L., YIN X., LYNN K.M., ROSS B.N., FAIR M., DAMRA M., SCIORILLO A.C., LIU Q., JACOBSON J.M., MOUNZER K., KOSTMAN J.R., ABDEL-MOHSEN M., MONTANER L.J, & PAPASAVVAS E. (2020). HCV modulates IgG glycosylation in HIV co-infected antiretroviral therapy suppressed individuals. AIDS (London, England), 34(10), 1461-1466.

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Cell Sorting and Telomere Analysis

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Cell sorting was performed using a MoFlo cell sorter equipped with SUMMIT software (Beckman Coulter, Buckinghamshire, UK). Following collagenase digestion of UC tissue, 2×10
⁶ cells were split into two tubes. One tube was stained with 5 μl of neat CD45-FITC (#F0861), CD235α-FITC (#F0870) (both from DAKO, Cambridge, UK), CD146-PE (BD Pharmingen) and CD31-APC (#130-092-652, Miltenyi Biotec), whereas the other was stained with 2.5 μl of neat isotype controls IgG1-FITC (#550617, BD Pharmingen), IgG1-PE (#MCA928PE, Serotec) and IgG1-APC (#130-098-846, Miltenyi Biotec). After incubation with relevant antibodies and washes, 2 μg/ml 7-aminoactinomycin D (7-AAD) (#A1310, Invitrogen) was added to exclude dead cells before sorting into four fractions: haemopoietic cell fraction (HC), CD45
⁺CD235α
⁺CD31
^-; EC fraction, CD45
^-CD235α
^-CD31
⁺; candidate MSC fraction, CD45
^-CD235α
^-CD31
^-CD146
⁺ and non-MSC fraction, CD45
^-CD235α
^-CD31
^-CD146
^-. The latter two subsets were processed for telomere length analysis.

Kouroupis D., Churchman S.M., McGonagle D, & Jones E.A. (2014). The assessment of CD146-based cell sorting and telomere length analysis for establishing the identity of mesenchymal stem cells in human umbilical cord. F1000Research, 3, 126.

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Identification of Cancer Stem Cell Markers

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We used the following antibodies to detect the expression of cancer stem cell (CSC) markers: anti-CD44-PE, anti-CD24-APC, anti-epithelial-specific antigen (ESA)-PE-Vio770, IgG1-PE, IgG1-APC, and IgG1-PE-Vio770 (Miltenyi Biotec, Bergisch Gladbach, Germany). Cells were harvested by digestion with trypsin without EDTA to produce a single-cell suspension. The cells were pelleted by centrifugation, washed twice with phosphate-buffered saline (PBS) and then incubated with the appropriate antibody in the dark for 10 min at 4°C. The appropriate isotype control antibodies were used at the same concentrations according to the manufacturer's instructions. After two washes with PBS, the samples were resuspended in 500 μl of PBS and analyzed with a BD FACSCalibur flow cytometer (BD Biosciences, San Jose, CA, USA) within 1 h.

Wang F., Rong L., Zhang Z., Li M., Ma L., Ma Y., Xie X., Tian X, & Yang Y. (2020). LncRNA H19-Derived miR-675-3p Promotes Epithelial-Mesenchymal Transition and Stemness in Human Pancreatic Cancer Cells by targeting the STAT3 Pathway. Journal of Cancer, 11(16), 4771-4782.

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