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Anti human igg pe

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Anti-human IgG-PE is a labeling reagent that consists of phycoerythrin (PE) conjugated to an antibody specific for human immunoglobulin G (IgG). This product is designed for the detection and quantification of human IgG in various applications.

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Lab products found in correlation

Anti human igm alexa fluor 647, by Southern Biotech (3 mentions) Anti human iga fitc, by Southern Biotech (3 mentions) Streptavidin pe, by Thermo Fisher Scientific (2 mentions) Polyjet in vitro dna transfection reagent, by SignaGen (1 mentions) Anti human cd3 apc h7, by BD (1 mentions) Anti human igg pe, by BD (1 mentions) Facscanto 2, by BD (1 mentions) Accuri, by BD (1 mentions) A549 cell, by American Type Culture Collection (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Hepes, by Lonza (1 mentions) Facsdiva, by BD (1 mentions) P1110, by Solarbio (1 mentions) Alexa fluor 647, by Thermo Fisher Scientific (1 mentions) Ovalbumin (ova), by Merck Group (1 mentions) Igg3 pe, by Southern Biotech (1 mentions) Igg1 pe, by Southern Biotech (1 mentions) Zeba spin desalting column, by Thermo Fisher Scientific (1 mentions) Magnetic beads, by Bio-Rad (1 mentions) Biotin pe, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

PE)-conjugated mouse anti-human IgG Fc antibody PE-conjugated goat anti-human IgG polyclonal antibodies Goat F(ab')2 Anti-Human IgG-PE Mouse anti-human IgG-PE Goat anti-human IgG PE Ab Goat anti-human IgG (γ-chain specific)-PE conjugated secondary antibody PE-conjugated goat F(ab)’2 anti-human IgG Goat anti-human IgG PE antibody Anti-human IgG Fc PE PE-conjugated goat anti-human IgG

13 protocols using anti human igg pe

1

Multiplex Assay for Analyzing SARS-CoV-2 RBD Antibody Binding

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A multiplex array consisting of reported RBD variants (n = 24) was used to examine the epitope resilience of human mAbs as previously described (Pymm et al., 2021 ). Briefly RBD variant multiplex bead cocktails were generated. Bead cocktails (1000 beads of each region per well) and mAbs were added at 8-fold 1:4 titrations, from a starting concentration of 80 nM per well into 384 well plates. Plates were incubated for 2h with shaking, washed twice with 0.05% PBS Tween and relative mAb binding was detected using anti-human IgG-PE (#9040-09, Southern Biotech) at 1.3 ug/ml for 2 hours with shaking.
Relative ACE2 inhibition was conducted using the same RBD variant multiplex bead cocktail and mAbs titrated, as described above, with the addition of 25 ug/ml biotinylated ACE2 per well. Plates were incubated for 2 h with shaking, washed twice with 0.05% PBS Tween followed by the addition of Streptavidin-PE (#S866, Thermo Fisher) and PE Biotin-XX conjugate (#P811, Thermo Fisher) for another 1 h with shaking. Plates were acquired on a Flexmap3D (Luminex Corporation). The relative binding of mAb or ACE2, was detected as phycoerythrin-labeled reporter and is measured as MFI (Median Fluorescence Intensity)
Wheatley A.K., Pymm P., Esterbauer R., Dietrich M.H., Lee W.S., Drew D., Kelly H.G., Chan L.J., Mordant F.L., Black K.A., Adair A., Tan H.X., Juno J.A., Wragg K.M., Amarasena T., Lopez E., Selva K.J., Haycroft E.R., Cooney J.P., Venugopal H., Tan L.L., O Neill M.T., Allison C.C., Cromer D., Davenport M.P., Bowen R.A., Chung A.W., Pellegrini M., Liddament M.T., Glukhova A., Subbarao K., Kent S.J, & Tham W.H. (2021). Landscape of human antibody recognition of the SARS-CoV-2 receptor binding domain. Cell Reports, 37(2), 109822.
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2

Binding Affinity and Competition Assay for SARS-CoV-2 RBD Mutants

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SARS-CoV-2 RBD mutants were purified and coupled onto magnetic multiplex beads (Luminex Corporation) as previously described and mixed in a bead cocktail (35 (link)). Bead cocktails were dispensed into each well of a black, clear-bottom, 384-well plate (Greiner), and mAbs at a starting concentration of 80 nM serially diluted were next added to the bead cocktail and incubated on a plate shaker for 2 hours at room temperature. The binding affinities of mAbs to different SARS-CoV-2 RBD mutants were detected using anti–human IgG-PE (Southern Biotech), and the mean fluorescence intensity (MFI) of the PE reporter was acquired on a FLEXMAP 3D machine (Luminex Corporation). To measure the competition of mAb-RBD binding by ACE2, bead cocktail and serially diluted mAbs were first mixed as described above, followed by the addition of Avi-tagged biotinylated ACE2. The plate was incubated on a plate shaker for 2 hours at room temperature. ACE2 binding was detected with streptavidin, R-PE conjugate (S866, Thermo Fisher Scientific) for 1 hour, followed by the addition of the R-PE Biotin-XX conjugate (P811, Thermo Fisher Scientific) for another 1 hour. The binding of ACE2 was detected as the MFI of the PE reporter using FLEXMAP 3D machine (Luminex Corporation).
Chia W.N., Tan C.W., Tan A.W., Young B., Starr T.N., Lopez E., Fibriansah G., Barr J., Cheng S., Yeoh A.Y., Yap W.C., Lim B.L., Ng T.S., Sia W.R., Zhu F., Chen S., Zhang J., Kwek M.S., Greaney A.J., Chen M., Au G.G., Paradkar P.N., Peiris M., Chung A.W., Bloom J.D., Lye D., Lok S, & Wang L.F. (2023). Potent pan huACE2-dependent sarbecovirus neutralizing monoclonal antibodies isolated from a BNT162b2-vaccinated SARS survivor. Science Advances, 9(30), eade3470.
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3

HIV-1 Env Gene Transfection and Antibody Staining

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293 T cells were transfected with 1 ug of plasmid DNA encoding an HIV-1 env gene (PolyJet™ In Vitro DNA Transfection Reagent (SignaGen Laboratories, Cat. No. SL100688) in 6-well plates and harvested at 48 h. Cells were split into separate staining reactions for monoclonal antibodies 3BNC117 (Cat. No. ARP-12474), 10-1074 (Cat. No. ARP-12477), 10E8 (Cat. No. ARP-12294), 447-52D (Cat. No. ARP-4030, all from BEI resources), b12 (Polymun, Cat. No. AB011), PGDM140015 (link) and PGT15193 (link) (both kindly provided by Dr. Devon Sok and International AIDS Vaccine Initiative). 2G12 was produced in Expi293F cells from publicly available heavy and light chain sequences94 (link) and purified using Protein A affinity chromatography. Cells were washed and stained with anti-human IgG-PE at 1:250 dilution (SouthernBiotech, Cat. No. 2040-09) for 15 min and fixed in 0.5% paraformaldehyde. Fluorescence was measured on a FACSymphony A5 SE Cell Analyzer and analyzed in FlowJo v.9.9.6.
Bai H., Lewitus E., Li Y., Thomas P.V., Zemil M., Merbah M., Peterson C.E., Thuraisamy T., Rees P.A., Hajduczki A., Dussupt V., Slike B., Mendez-Rivera L., Schmid A., Kavusak E., Rao M., Smith G., Frey J., Sims A., Wieczorek L., Polonis V., Krebs S.J., Ake J.A., Vasan S., Bolton D.L., Joyce M.G., Townsley S, & Rolland M. (2024). Contemporary HIV-1 consensus Env with AI-assisted redesigned hypervariable loops promote antibody binding. Nature Communications, 15, 3924.
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4

Chimeric Antigen Receptor T-Cell Phenotyping

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To determined transduction efficiency, surface CAR expression was evaluated by flow cytometry (FACSCanto II or Accuri, BD Biosciences) by staining transduced cells with anti-human IgG-phycoerythrin (PE) (SouthernBiotech) and CD3-allophycocyanin (APC) (Miltenyi Biotec), respectively. For phenotyping, CAR-transduced cells were stimulated with PSMA+ tumor cells (C4-2 or LNCaP) for 24 h at an E:T ratio of 1:1, before cells were harvested and evaluated. Activation was evaluated based on the expression of CD69 or CD25 (CD69-APC, clone CH4; CD25-PE, clone 3G10; both Thermo Fisher Scientific). T cell subsets were determined by staining cells with anti-human CD62L-Brilliant Violet 421 (BV421) (BD Biosciences), anti-human CD45RA-fluorescein isothiocyanate (FITC) (BioLegend), anti-human CD3-APC/H7 (BD Biosciences), and anti-human IgG-PE (SouthernBiotech). To determine exhaustion, cells were stained with anti-human CD279-FITC (PD-1, BD Biosciences), anti-human CD223-eFluor 710 (LAG-3, BD Biosciences), anti-human CD3-APC/H7, and anti-human IgG-PE.
Alzubi J., Dettmer-Monaco V., Kuehle J., Thorausch N., Seidl M., Taromi S., Schamel W., Zeiser R., Abken H., Cathomen T, & Wolf P. (2020). PSMA-Directed CAR T Cells Combined with Low-Dose Docetaxel Treatment Induce Tumor Regression in a Prostate Cancer Xenograft Model. Molecular Therapy Oncolytics, 18, 226-235.
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5

SARS-CoV-2 Antibody Profiling Assay

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293F-spike–expressing cells were incubated with 100-fold diluted plasma samples for 30 min at 37°C. Cells were washed twice and stained with anti-human IgG PE, anti-human IgM Alexa Fluor 647, and anti-human IgA FITC (all from SouthernBiotech). Cells were then fixed with 4% formaldehyde solution, and fluorescence was evaluated on an LSR II (BD Biosciences).
Yu J., Thomas P.V., McMahan K., Jacob-Dolan C., Liu J., He X., Hope D., Martinez E.J., Chen W.H., Sciacca M., Hachmann N.P., Lifton M., Miller J., Powers O.C., Hall K., Wu C., Barrett J., Swafford I., Currier J.R., King J., Corbitt C., Chang W.C., Golub E., Rees P.A., Peterson C.E., Hajduczki A., Hussin E., Lange C., Gong H., Matyas G.R., Rao M., Paquin-Proulx D., Gromowski G.D., Lewis M.G., Andersen H., Davis-Gardner M., Suthar M.S., Michael N.L., Bolton D.L., Joyce M.G., Modjarrad K, & Barouch D.H. (2022). Protection against SARS-CoV-2 Omicron BA.1 variant challenge in macaques by prime-boost vaccination with Ad26.COV2.S and SpFN. Science Advances, 8(47), eade4433.
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6

FACS Analysis of cMet Antibody Binding

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FACS analysis was performed using a BD FACSDiva instrument (version 8.0, BD Biosciences) to verify that the cMet Ab could bind cMet. A549 cells (ATCC) were used. Cells were cultured using F12 HAMS media supplemented with 10% FBS (Gibco), 1% P/X, 2% HEPES and EGM (Lonza, CC-3124). For the experiment, A549 cells (5 × 105) were added to a 1.5 ml tube, and the culture was washed out. Then, 4 µl of cMet Ab (Abcam) was diluted in 100 µl of PBS, after which this solution was reacted with the cell suspension at 4 °C for 30 minutes. Afterward, the cells were centrifuged at 4000 rpm for 3 minutes and washed twice. Then, 2 µl of anti-human IgG-PE (Southern Biotech) was added to the suspension, and the cells were washed, resuspended in 1 ml of PBS, and then subjected to FACS analysis.
Kim Y.C., Lee J., An J.N., Kim J.H., Choi Y.W., Li L., Kwon S.H., Lee M.Y., Lee B., Jeong J.G., Yu S.S., Lim C.S., Kim Y.S., Kim S., Yang S.H, & Lee J.P. (2019). Renoprotective effects of a novel cMet agonistic antibody on kidney fibrosis. Scientific Reports, 9, 13495.
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7

Quantifying GITR-IBI37G5 Binding Kinetics

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3 × 105 PBMCs or Jurkat-GITR reporter cells were seeded onto 96-well plates and incubated with different concentration of IBI37G5 at 4°C for 1 h. Cells were then fixed with 4% fixative solution (Solarbio, P1110). Recombinant GITR protein (Acrobiosystems, GIR-H5228) conjugated with Alexa Fluor 647(Thermo Fisher, A20173)and anti-human IgG-PE (Southern biotech, 2040-09) were used to stain freely exposed IBI37G5 binding sites and cell surface-bound IBI37G5, respectively. Fluorescent signals were detected by flow cytometry.
Liu H., Wu W., Sun G., Chia T., Cao L., Liu X., Guan J., Fu F., Yao Y., Wu Z., Zhou S., Wang J., Lu J., Kuang Z., Wu M., He L., Shao Z., Wu D., Chen B., Xu W., Wang Z, & He K. (2022). Optimal target saturation of ligand-blocking anti-GITR antibody IBI37G5 dictates FcγR-independent GITR agonism and antitumor activity. Cell Reports Medicine, 3(6), 100660.
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8

Multiplex Antibody Profiling of Pertussis Immunity

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Pertussis antigen-specific antibody responses were quantified in human plasma by performing an indirect serological assay with xMAP Microspheres (details described in xMAP Cookbook, Luminex 5th edition). Pertussis, Tetanus, and Diphtheria antigens (PT, PRN, Fim2/3, TT, and DT (all from List Biological Laboratories) and FHA (Sigma) and as a negative control Ovalbumin (Sigma) were coupled to uniquely coded beads (xMAP MagPlex Microspheres, Luminex Corporation). PT was inactivated by incubation with 1% formaldehyde (PFA) at 4°C for 1 h. 1% PFA PT and TT were then purified using Zeba spin desalting columns (ThermoFisher). The antigens were coupled with each unique conjugated microsphere using the xMAP Antibody Coupling Kit (Luminex Corporation). Plasma was mixed with a mixture of each conjugated microsphere, and WHO International Standard Human Pertussis antiserum was used as a reference standard (NIBSC, 06/140). Subsequently, the mixtures were washed with 0.05% TWEEN 20 in PBS (Sigma-Aldrich) to exclude non-specific antibodies, and targeted antibodies responses were detected via anti-human IgG-PE, IgG1-PE, IgG2-PE, IgG3-PE, IgG4-PE (all from SouthernBiotech) and human IgE-PE (ThermoFisher). Samples were subsequently measured on an FLEXMAP 3D instrument (Luminex Corporation), and the log(10) of the median fluorescent intensity (MFI) was calculated.
Shinde P., Soldevila F., Reyna J., Aoki M., Rasmussen M., Willemsen L., Kojima M., Ha B., Greenbaum J.A., Overton J.A., Guzman-Orozco H., Nili S., Orfield S., Gygi J.P., da Silva Antunes R., Sette A., Grant B., Olsen L.R., Konstorum A., Guan L., Ay F., Kleinstein S.H, & Peters B. (2024). A multi-omics systems vaccinology resource to develop and test computational models of immunity. Cell Reports Methods, 4(3), 100731.
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9

Magnetic Bead-Based Antibody Quantification

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A magnetic bead-based assay (Brown et al., 2012 (link)) was used to measure antibody expression and biodistribution. Beads were conjugated to anti-human antigen-binding fragment (Fab) to capture mAbs of interest. Briefly, anti-human IgG F(ab’)2 fragment (109-006-097; Jackson Immune Research) was conjugated to fluorescent microspheres (MagPlex-C Microspheres, Luminex Corp.) at a ratio of 6.5 μg protein/100 μl microspheres. Samples were diluted as indicated in mAb expression and biodistribution figures and then incubated with microspheres (500–750 beads/well) overnight at 4°C and washed in PBS with 1% BSA, 0.05% Tween-20, and 0.1% sodium azide. Anti-human IgG PE (2040-09; Southern Biotech) was incubated at 0.65 μg/ml for 45 min in PBS-TBN. The microspheres were washed and resuspended in 90 μl of sheath fluid (Luminex) and read using a Bio-plex array reader (FlexMap 3D, OR MAGPIX). The median fluorescence intensity of the PE signal was determined for each sample at indicated dilutions. For biodistribution assessment, the signal is reported as the fold increase in PE signal in treated pups relative to untreated controls.
Backes I.M., Byrd B.K., Slein M.D., Patel C.D., Taylor S.A., Garland C.R., MacDonald S.W., Balazs A.B., Davis S.C., Ackerman M.E, & Leib D.A. (2022). Maternally transferred mAbs protect neonatal mice from HSV-induced mortality and morbidity. The Journal of Experimental Medicine, 219(12), e20220110.
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10

SARS-CoV-2 S Protein Expression and Antibody Binding Assay

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SARS-CoV-2 S-expressing FreeStyle 293F cells were generated by transfection with linearized plasmid encoding a codon-optimized full-length SARS-CoV-2 S protein matching the amino acid sequence of the IL1/2020 isolate (GenBank, MN988713). Stable transfectants were single-cell sorted and selected to obtain a high-level spike surface expressing clone (293F-Spike-S2A). 293F-Spike-S2A cells were incubated with mAbs diluted threefold from 15 to 0.06 µg ml−1 for 30 min at 37 °C. Cells were washed twice and stained with anti-human IgG PE (Southern Biotech). Cells were then fixed with 4% formaldehyde solution and fluorescence was evaluated on an LSR II (BD Bioscience).
Dussupt V., Sankhala R.S., Mendez-Rivera L., Townsley S.M., Schmidt F., Wieczorek L., Lal K.G., Donofrio G.C., Tran U., Jackson N.D., Zaky W.I., Zemil M., Tritsch S.R., Chen W.H., Martinez E.J., Ahmed A., Choe M., Chang W.C., Hajduczki A., Jian N., Peterson C.E., Rees P.A., Rutkowska M., Slike B.M., Selverian C.N., Swafford I., Teng I.T., Thomas P.V., Zhou T., Smith C.J., Currier J.R., Kwong P.D., Rolland M., Davidson E., Doranz B.J., Mores C.N., Hatziioannou T., Reiley W.W., Bieniasz P.D., Paquin-Proulx D., Gromowski G.D., Polonis V.R., Michael N.L., Modjarrad K., Joyce M.G, & Krebs S.J. (2021). Low-dose in vivo protection and neutralization across SARS-CoV-2 variants by monoclonal antibody combinations. Nature Immunology, 22(12), 1503-1514.
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