PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
Overview
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Product name
PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® -
Product overview
R-PE Conjugation Kit/ R-PE Labeling Kit ab102918 uses a simple and quick process for PE labeling / conjugation of antibodies. It can also be used to conjugate other proteins or peptides. Learn about our antibody labeling kits and their advantages.
To conjugate an antibody to R-PE using this kit:
- add modifier to antibody and incubate for 3 hrs
- add quencher and incubate for 30 mins
The PE conjugated antibody can be used immediately in WB, ELISA, IHC etc. No further purification is required and 100% of the antibody is recovered for use.
Learn about buffer compatibility below; for incompatible buffers and low antibody concentrations, use our rapid antibody purification and concentration kits. Use the FAQ to learn more about the technology, or about conjugating other proteins and peptides to R-PE.
Custom size conjugation kits up to 100 mg are available on demand. Please contact us to discuss your requirements.
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Notes
This product is manufactured by Expedeon, an Abcam company, and was previously called Lightning-Link® R-PE Labeling Kit. 703-0004 is the same as the 3 mg size. 703-0003 is the same as the 5 x 600 ug size. 703-0010 is the same as the 3 x 60 ug size. 703-0030 is the same as the 3 x 10 ug size. 703-0015 is the same as the 600 µg size. 703-0005 is the same as the 60 µg size.
Amount and volume of antibody for conjugation to R-PE
Kit size Recommended
amount of antibodyMaximum antibody
volume13 x 10 µg 3 x 10 µg 3 x 10 µL 60 µg 1 x 60 µg 1 x 60 µL 3 x 60 µg 3 x 60 µg 3 x 60 µL 600 µg 1 x 600 µg 1 x 600 µL 5 x 600 µg 5 x 600 µg 5 x 600 µL 3 mg 1 x 3 mg 1 x 3 mL The selling size of this product is now based on the amount of antibody that can be conjugated with the kit; the amount of antibody advised that can be used with the kit has also been updated to reflect what will give the best conjugation results. The quantity and formulation of reagents provided have not changed, if you have been previously using the kit successfully with a different amount of antibody, there is no need to change the way that you are using the kit.
1Ideal antibody concentration is 1mg/ml. 0.5 - 1 mg/ml can be used if the maximum antibody volume is not exceeded. Antibodies > 1 mg/ml or < 0.5 mg/ml should be diluted /concentrated.
Buffer Requirements for Conjugation
Buffer should be pH 6.5-8.5.
Compatible buffer constituents
If a concentration is shown, then the constituent should be no more than the concentration shown. If several constituents are close to the limit of acceptable concentration, then this can inhibit conjugation.50mM / 0.6% Tris1 0.1% BSA 50% glycerol 0.1% sodium azide PBS Potassium phosphate Sodium chloride HEPES Sucrose Sodium citrate EDTA Trehalose 1 Tris buffered saline is almost always ≤ 50 mM / 0.6%
Incompatible buffer constituents
Thiomerosal Proclin Glycine Arginine Glutathione DTT If a constituent of the buffer containing your antibody or protein is not listed above, please check the FAQ or contact us.
Only purified antibodies are suitable for use, ie. where other proteins, peptides, or amino acids are not present: antibodies in ascites fluid, serum or hybridoma culture media are incompatible.
Storing and handling conjugation kits
Lyophilized Lightning-Link® components are hygroscopic.
Kits are intentionally shipped at ambient temperature with silica gel to avoid exposure to moisture. Upon receipt, store the kit frozen and protect from moisture. Before opening the outer container, allow the lyophilized components to reach room temperature to minimize condensation.
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components 60 µg 600 µg 5 x 600 µg 3 x 60 µg 3 x 10 µg 1 x 3 mg ab274106 - Modifier reagent 1 x 200µl 1 x 200µl 1 x 1200µl 1 x 200µl 1 x 200µl 1 x 1200µl ab274296 - Quencher reagent 1 x 200µl 1 x 200µl 1 x 1200µl 1 x 200µl 1 x 200µl 1 x 1200µl ab274122 - R-PE mix 1 x 60µg 1 x 600µg 5 x 600µg 3 x 60µg 3 x 10µg 1 x 3mg -
Research areas
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Alternative names
- R Phycoerythrin
- RPE
Associated products
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Related Products
Images
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Flow Cytometry - PE / R-Phycoerythrin Conjugation Kit Lightning-LinkImage from Robinson, Andrew P., et al., PloS one, 9(9):e107649. doi: 10.1371/journal.pone.0107649. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/
Robinson, Andrew P., et al used PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918) as part of characterizing oligodendroglial populations. They used the kit to conjugate PE / R-Phycoerythrin to Mouse monoclonal anti-GALC antibody for use in flow cytometry.
SJL/J mice were immunized with PLP139-151 and scored daily for clinical disease. A cohort of SJL/J mice was sacrificed, and spinal cords were analyzed by flow cytometry (n=5). (A) Cells were distinguished from debris by forward and side scatter then singlet cells were gated. Live cells were gated by dead cell exclusion, and CNS resident cells were identified as CD45- or CD45low. (B) Oligodendroglial cells were defined by double positive staining: A2B5+PDGFRalpha+ early OPCs, A2B5+NG2+ intermediate OPCs, NG2+O4+ late OPCs, O4+MOG+ pre-myelinating oligodendrocytes, and GALC+MOG+ mature oligodendrocytes. -
Conjugation - R-Phycoerythrin Conjugation (ab102918) -
PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® labeling ADRB2 antibody for Flow CytometryImage from Bigler MB et al., PloS One, 10(12):e0145635. Fig 3.; doi: 10.1371/journal.pone.0145635. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/Bigler MB et al. used ab102918 to assess expression of the epinephrine receptor (ADRB2) on CD3- CD56+ lymphocytes by flow cytometry.
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PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® labeling F(ab')2 antibody for Bead arrayImage from Ayoglu B et al., PLoS One 9(5):e96403. Fig 2.; doi: 10.1371/journal.pone.0096403. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/Ayoglu B et al. used ab102918 to detect complement activation driven C3 deposition on beads.
For measuring the background, classical, lectin & alternative and only alternative pathway activation, a serially diluted serum sample (1∶1–1∶160) was applied to empty, human IgG. The assay buffer for serum dilution contained either Ca2+-Mg2+, which promotes complement activation, or EDTA, which blocks complement activation. Plot displays for each serum dilution the respective median fluorescence intensity (MFI) value against varying concentrations of human IgG.
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PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® labeling anti-MUC1 SP antibodies FACSImage from Kovjazin R et al., PLoS One, 9(1):e85400. Fig 3.; doi: 10.1371/journal.pone.0085400. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/Kovjazin R et al. used ab102918 to evaluate the cell surface presence of the MUC1 SP domain by FACS.
The cell surface presence of the MUC1 SP domain was evaluated by gated FACS analysis on PC cells in fresh BM aspirates obtained from MM patients (A–E) and normal naive sample (F–H).
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PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® labeling SPmAb-6 antibody for Imagestream analysisimage from Kovjazin R et al., PLoS One, 9(1):e85400. Fig 2.; doi: 10.1371/journal.pone.0085400. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/Kovjazin R et al. used ab102918 for detection of MUC1 SP on the membrane of MUC1-positive tumor cells.
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PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® labeling a panel of antibodies for Microsphere immunoassay based on LuminexImage from Charlermroj R et al., PloS One, 8(4):e62344. Fig 2.; doi: 10.1371/journal.pone.0062344. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/Charlermroj R et al. used ab102918 as part of an experiment studying four different plant pathogens: a fruit blotch bacterium Acidovorax avenae subsp. citrulli (Aac), chilli vein-banding mottle virus (CVbMV, potyvirus), watermelon silver mottle virus (WSMoV, tospovirus serogroup IV) and melon yellow spot virus (MYSV, tospovirus).
(A-G) X-axis is antibody-coated microsphere and y-axis is median fluorescent intensity (MFI) from each RPE-labeled antibody. (H) Summary of selected antibody pair sets for the detection of the four plant pathogens.
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PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® labeling A20 antibody for FACSImage from Hjelmeland AB et al., PLoS Biol., 8(2):e1000319. Fig 2.; doi: 10.1371/journal.pbio.1000319.Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/Hjelmeland AB et al. used ab102918 as part of an experiment studying the mechanisms that cause A20 regulator to have differential effects on tumor growth and cancer cell behaviors.
They used the PE labelling kit to conjugate phycoerythrin to A20 antibody for use in FACS.
FACS plots are shown for T08-837 (A) and CW468 (B).
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PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® labeling anti-mouse insulin antibody for Flow CytometryImage from Kalis, M et al., PLoS One, 6(12):e29166. Fig 5.; doi: 10.1371/journal.pone.0029166. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/Kalis M et al. used ab102918 as part of examining beta-cells deletion of Dicer1.
They used the kit to conjugate PE to anti-mouse insulin antibody for use in flow cytometry.
Islet cells suspension from 50 islets from 8 weeks old littermates and RIP-Cre Dicer1Δ/Δ mice were stained for insulin and glucagon and analyzed by flow cytometry. Side scatter (SSC) and forward scatter (FSC) analysis of islet cells and the respective gate used to identify live cells. Littermate control islet cells contained significantly more insulin-positive cells (P<0.001) and less glucagon-positive cells (P<0.05) than the RIP-Cre Dicer1Δ/Δ islet cells. Representative of 3 independent experiments.
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PE conjugation kit used to label DDR2 antibody for Flow CytometryBagchi RA et al. BMC Biol. 2016 Mar 17;14:21 Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/RA Bagchi et al used PE conjugation kit / PE labeling kit ab102918 as part of examining the conversion of fibroblasts to myofibroblasts. They used the PE labeling kit to conjugate phycoerythrin to a DDR2 antibody for use in flow cytometry.
Charts are forward-scatter plots illustrating flow cytometry analysis of cardiac cells from WT and scleraxis KO mice. Left column, unstained cells; center column, stained cells from WT tissue; right column, stained cells from scleraxis KO tissue. Results are representative of assessments from n = 3 independent tissue samples. Purple outline denotes labeled cells, and is derived from unstained plots.
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PE conjugation to an anti-chymase antibody with ab102918 PE Conjugation KitImage from Rönnberg E et al., Frontiers in immunology., 10, 1361.. Fig 4C.; doi: 10.3389/fimmu.2019.01361 Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/.Ronnberg E et al. conjugated PE to an anti-chymase antibody with ab102918 PE Conjugation Kit as part of investigating the role of IL-33 in mast cells.
The mean fluorescence intensity of chymase expression in cells was measured by intra-cellular flow cytometry staining and normalized to the respective isotype control.This data shows that there was no significant change in the expression on chymase in cord blood-derived mast cells (CBMC) when treated with different combinations of IL-33 and TSLP.
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Conjugation of recombinant influenza A H1N1 NP protein with PE or APC fluorochromes using ab102918Image from Tan HX et al., Frontiers in immunology., 10, 611.. Fig 4A.; doi: 10.3389/fimmu.2019.00611 Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/Tan H-X et al. labeled recombinant influenza A H1N1 NP protein with PE or APC fluorochromes using ab102918 PE / R-Phycoerythrin Conjugation Kit and ab201807 APC Conjugation Kit.
Representative HA and NP probe staining in flow cytometry of GC B cells (B220+ IgD- CD38lo GL7+) isolated from lung inducible bronchus-associated tissues (iBALT), mediastinal LN (MLN), and spleen of mice at d35 post-infection with A/Puerto Rico/8/34 (PR8).
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Flow cytometry using antibody conjugated using EasyLink R-Phycoerythrin conjugation kit (ab102918)This image is courtesy of an anonymous Abreview.Flow cytometry histogram showing integrin beta-3 positive population of platelets from wildtype and knockout mice. Integrin beta-3 antibody was conjugated using EasyLink R-Phycoerythrin conjugation kit (ab102918). Flow cytometry was performed using platelets from wild type and integrin beta-3 knockout mice. Mice that expressed beta-3 (shown in red) had a clear shift in FL-2 fluorescence over beta-3 knockout mice (shown in black).
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (93)
ab102918 has been referenced in 93 publications.
- Guito JC et al. Asymptomatic Infection of Marburg Virus Reservoir Bats Is Explained by a Strategy of Immunoprotective Disease Tolerance. Curr Biol 31:257-270.e5 (2021). PubMed: 33157026
- Bonser LR et al. Flow-Cytometric Analysis and Purification of Airway Epithelial-Cell Subsets. Am J Respir Cell Mol Biol 64:308-317 (2021). PubMed: 33196316
- van der Donk LEH et al. Separate signaling events control TCR downregulation and T cell activation in primary human T cells. Immun Inflamm Dis 9:223-238 (2021). PubMed: 33350598
- Tan TK et al. A COVID-19 vaccine candidate using SpyCatcher multimerization of the SARS-CoV-2 spike protein receptor-binding domain induces potent neutralising antibody responses. Nat Commun 12:542 (2021). PubMed: 33483491
- Zhang C et al. NLRP3 inflammasome induces CD4+ T cell loss in chronically HIV-1-infected patients. J Clin Invest 131:N/A (2021). PubMed: 33720048