Pe anti rabbit igg

Manufactured by BioLegend

PE-anti-rabbit IgG is a secondary antibody conjugated with the fluorescent dye Phycoerythrin (PE). It is designed to detect and bind to rabbit immunoglobulin G (IgG) antibodies in various immunoassays and research applications.

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PE-IgG (4 citations)

6 protocols using pe anti rabbit igg

Multiparametric Flow Cytometry Analysis

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To examine PD-1, CTLA-4 and GRAIL expression, splenocytes or CD4 T cells from control and infected animals or in vitro infected CD4 T cells were washed with saline solution 2% FBS and incubated with anti-mouse CD32/CD16 antibody for 20 minutes at 4°C to block Fc receptors. Then, cells were incubated with APC labeled anti-CD4, PercP labeled anti-CD3 (BD Pharmingen), and with PE-labeled anti-PD-1 or anti-CTLA-4 (BD Pharmingen) for 20 min at 4°C.
For the assessment of intracellular GRAIL expression, cells were first stained with FITC-CD3 and APC-CD4 antibodies, and then fixed and permeabilized with Citofix/Citoperm (BD Biosciences,) for 30 min followed by reacting with rabbit anti-GRAIL primary Ab (Abcam) for 45 min. After washing, cells were stained for 20 min with PE–anti-rabbit IgG (Biolegend). Finally, cells were washed twice with saline solution of 2% FBS, and stored at 4°C in the dark until analysis using a FACS flow cytometer (FACS Canto II, BD Biosciences). The results were processed using Flow Jo software (version 7.6.2).

Stempin C.C., Rojas Marquez J.D., Ana Y, & Cerban F.M. (2017). GRAIL and Otubain-1 are Related to T Cell Hyporesponsiveness during Trypanosoma cruzi Infection. PLoS Neglected Tropical Diseases, 11(1), e0005307.

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Phage Staining of Peptide-Pulsed T2 Cells

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Phage staining of peptide-pulsed T2 cells was performed with 50 μl phage supernatant on ice for 1 hour, followed by staining with 1 μg of rabbit anti-M13 antibody (Novus Biologicals, NB100-1633) and PE anti-rabbit IgG (BioLegend, 406421). HLA-A*02 staining was performed by staining cells with APC-labeled anti-human HLA-A*02 (BB7.2, BioLegend, 343308) or mouse isotype IgG2b, κ (BioLegend, 402206). Human T cells engrafted in mice were stained with Brilliant Violet (BV) 421 anti-human CD3 (SK7, BioLegend, 344834) and BV605 anti-human CD8(SK1, BioLegend, 344742). Stained cells were analyzed using an LSRII flow cytometer (Becton Dickinson) or an iQue Screener (IntelliCyt).

Hsiue E.H., Wright K.M., Douglass J., Hwang M.S., Mog B.J., Pearlman A.H., Paul S., DiNapoli S.R., Konig M.F., Wang Q., Schaefer A., Miller M.S., Skora A.D., Azurmendi P.A., Murphy M.B., Liu Q., Watson E., Li Y., Pardoll D.M., Bettegowda C., Papadopoulos N., Kinzler K.W., Vogelstein B., Gabelli S.B, & Zhou S. (2021). Targeting a neoantigen derived from a common TP53 mutation. Science (New York, N.Y.), 371(6533), eabc8697.

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Visualization of FlgE and EctoATP5B in HUVECs and SVECs

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HUVECs and SVECs cultured on glass coverslip were incubated with His-FlgE/His-FlgEM (10 μg/ml) and anti-ATP5B antibodies (5 μg/ml) for 1 h. The cells were fixed with 4% paraformaldehyde (PFA) for 20 min and blocked with 3% bovine serum albumin (BSA) for 30 min. FlgE was stained green with primary mouse anti-6× His antibodies (0.6 μg/ml, ProteinTech Group) at 4°C overnight and secondary fluorescein isothiocyanate (FITC)–anti-mouse IgG (eBioscience, San Diego, CA) for 1 h. EctoATP5B was stained red using phycoerythrin (PE)–anti-rabbit IgG (BioLegend, San Diego, CA) for 1 h. The images were observed under a confocal fluorescence microscope (Leica, Wetzlar, Germany).

Li Y., Shen Y., Zheng Y., Ji S., Wang M., Wang B., Han Q., Tian Y, & Wang Y. (2021). Flagellar Hook Protein FlgE Induces Microvascular Hyperpermeability via Ectopic ATP Synthase β on Endothelial Surface. Frontiers in Cellular and Infection Microbiology, 11, 724912.

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Quantifying Apoptosis in Draining LNs

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C57BL/6 or proxtom mice were immunized with 10 μg ova488, 2 μg poly(I:C), 2 μg αCD40 in the footpad and flank. Naive or mice immunized 2.5 weeks prior were killed and their popliteal or inguinal LNs placed in optimal cutting temperature compound or fixed in formalin and then paraffin embedded. About 7–10 μm sections were cut using a cryostat or microtome and stained with an antibody against LYVE-1 (1:400 Abcam polyclonal) or LYVE1-APC (1:50 R&D systems clone 223322) and cleaved caspase 3 (1:100, Cell Signaling). Secondary antibodies used were anti-rabbit IgG-PE (1:200 Biolegend) and anti-rabbit IgG-dylight647 (1:200 Biolegend). Sections were imaged using a Nikon Eclipse Ti series fluorescent microscope at ×20 magnification with an aperture of 0.50. Images were taken with a Photometrics Coolsnap Dyano fluorescent camera. Images were analyzed using NIS elements AR software to evaluate co-localization. Multiple LN sections were analyzed from five LNs of infected mice or two LNs from naive mice and the number of cells co-localized was quantified per LN.

Kedl R.M., Lindsay R.S., Finlon J.M., Lucas E.D., Friedman R.S, & Tamburini B.A. (2017). Migratory dendritic cells acquire and present lymphatic endothelial cell-archived antigens during lymph node contraction. Nature Communications, 8, 2034.

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Multiparameter Flow Cytometry Analysis

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Antibodies were purchased from Biolegend. The following primary antibodies were used: anti-CD16/CD32 (Biolegend, Cat#: 101301, clone: 93), anti-CD8-PE-cy7 (Biolegend, Cat#: 100721, clone: 53-6.7), anti-CD11c-FITC (Biolegend, Cat#: 117305, clone: N418) and anti-CD11b-Pacif blue (Biolegend, Cat#: 101223, clone: M1/70), anti-B220-APC-cy7 (Biolegend, Cat#: 103223, clone: RA3-6B2), anti-CD86-PE (Biolegend, Cat#: 105007, clone: GL-1), anti- H-2K^b bound to SIINFEKL-APC (Biolegend, Cat#:141605, clone:25-D1.16), anti-CD45.2-APC (Biolegend, Cat#:109814, clone:104), anti-CD45.2-APCcy7 (Biolegend, Cat#:109823, clone:104), anti-PD-L1-PE (Biolegend, Cat#:124307, clone:10F.9G2), isotype control-PE (Biolegend, Cat#:400607, clone:RTK4530), anti-F4/80-PE/cy7 (Biolegend, Cat#:123113, clone:BM8), anti-Gr-1-FITC (Biolegend, Cat#:108419, clone:RB608C5), anti-MHCII-BV605 (Biolegend, Cat#:107639, clone:M5/114.15.2), anti-rabbit IgG-PE (Biolegend, Cat#:406421, clone:poly4064). Flow data were acquired on a BD^™ LSR II flow cytometer and analyzed using Flowjo software.

Luo M., Wang H., Wang Z., Cai H., Lu Z., Li Y., Du M., Huang G., Wang C., Chen X., Porembka M.R., Lea J., Frankel A.E., Fu Y.X., Chen Z.J, & Gao J. (2017). A STING-Activating Nanovaccine for Cancer Immunotherapy. Nature nanotechnology, 12(7), 648-654.

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Multiparameter Flow Cytometry Analysis

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