ELISA - Biotinylation Kit / Biotin Conjugation Kit (Fast, Type A) - Lightning-Link® (ab201795)
Direct ELISA dose response curve generated with varying concentration of biotin conjugated antibodies. Anti-MMP1 antibody [EPR19769-152] - BSA and Azide free (Detector) (ab242835), Anti-EGF antibody [EPR19899-46] - BSA and Azide free (Detector) (ab242990),Anti-IL-5 antibody [EPR20012-256] - BSA and Azide free (Capture) (ab242955) and Anti-MMP9 antibody [EPR18543-47] - BSA and Azide free (Capture) (ab242950) were conjugated to biotin using Biotinylation Kit / Biotin Conjugation Kit (Fast, Type A) - Lightning-Link® (ab201795) to produce Conjugate A, B, C and Control respectively. Briefly, each well was coated with Recombinant Protein G. After blocking with 5% BSA blocking buffer for 2 hours, plates were coated with 1ug/ml of biotin conjugated antibodies. Detection was performed using a HRP-conjugated Streptavidin (ab7403), and visualised using a TMB substrate. Plates were read at 450nm using an Agilent BioTek Synergy HTX multimode reader.
Western blot - Biotinylation Kit / Biotin Conjugation Kit (Fast, Type A) - Lightning-Link® (ab201795)
Drachsler, Moritz, et al used Biotinylation Kit / Biotin Conjugation Kit (Fast, Type A) - Lightning-Link® (ab201795) as part of examining expression of CD95. They used the kit to conjugate biotin to anti-human CD95 for use in Western blot.
IP for CD95 and P-Tyrosine GSCs in naive, CD95-WT and CD95-mut GBM cells stimulated with CD95L-T4. Blots were probed with anti-P85 (regulatory PI3K subunit), anti-Sfk, anti-CD95 or anti-P-Tyrosine antibodies, respectively.
Image from Drachsler, Moritz, et al., Cell death & disease 7.4 (2016): e2209-e2209. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/
Multiplex Protein Detection - Biotinylation Kit / Biotin Conjugation Kit (Fast, Type A) - Lightning-Link® (ab201795)
Hähnel, Viola, et al used Biotinylation Kit / Biotin Conjugation Kit (Fast, Type A) - Lightning-Link® (ab201795) as part of examining cytokine levels of plasma donors. They used the kit to conjugate Biotinylation Kit / Biotin to cytokine proteins for use in multiplex protein detection.
Cytokines, lipopolysaccharide binding protein (LBP) and bactericidal permeability increasing protein (BPI) of convalescent donors were compared to healthy control group. The following cytokines were below limit of detection: IL1β, IL1RA, IL2, IL3, IL6, IL10, IL12p40, IL15, IL21, IL22, IL23, IFNβ, IFNγ, GM-CSF, MIP1α and TNFα.
Image from Hahnel, Viola, et al., PloS one, 15(12): e0243967; doi: 10.1371/journal.pone.0243967. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/
Flow Cytometry - Biotinylation Kit / Biotin Conjugation Kit (Fast, Type A) - Lightning-Link® (ab201795)
Flow Cytometry - Biotinylation Kit / Biotin Conjugation Kit (Fast, Type A) - Lightning-Link.
Giussani, Stefania, et al used Biotinylation Kit / Biotin Conjugation Kit (Fast, Type A) - Lightning-Link® (ab201795) as part of examining interaction of C3d ligand with complement interfering protein (CIP). They used the kit to conjugate biotin to C3d for use in flow cytometry.
Flow cytometry analysis of C3d interaction with B cells in presence or absence of Complement Interfering Protein (CIP). A) Biotinylated C3d was preincubated with SA and then with 1–9 μM of CIP, 9 μM of Fib3, or buffer alone. Each mixture was added to Raji B cells and treated as described. Binding of the biotinylated C3d–SA complex to cells was revealed by flow cytometry using a C3d-specific mAb and a phycoerythrin-labeled secondary antibody. Mean fluorescence intensity values of the peaks were analyzed by the FlowJo software. The graph shows the percentage of the residual mean fluorescence intensity values in the presence of a competitor compared with buffer alone, as derived from 4 independent experiments. B) Flow cytometry analysis of C3d binding to enriched B cells from human PBMCs in presence or absence of 1.5 or 9 μM CIP or 9 μM of Fib3; experimental conditions were the same as in A.
Image from Giussani, Stefania, et al., The FASEB Journal 33.3 (2019): 4448-4457. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/
Electron Microscopy - Biotinylation Kit / Biotin Conjugation Kit (Fast, Type A) - Lightning-Link® (ab201795)
Electron Microscopy - Biotinylation Kit / Biotin Conjugation Kit (Fast, Type A) Lightning-Link.
Hansen, Uwe, et al used Biotinylation Kit / Biotin Conjugation Kit (Fast, Type A) - Lightning-Link® (ab201795) as part of examining WARP-collagen IV interactions in in vitro in cartilage. They used the kit to conjugate biotin to recombinant WARP for use in Electron Microscopy.
Biotinylated recombinant WARP was visualized by 5 nm gold labeled with streptavidin. Scale bar is 25 nm
Image from Hansen, Uwe, et al., Plos one, 7(12): e52793.; doi: 10.1371/journal.pone.0052793. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/
Fluorescence Microscopy - Biotinylation Kit / Biotin Conjugation Kit (Fast, Type A) - Lightning-Link® (ab201795)
Fluorescence Microscopy - Biotinylation Kit / Biotin Conjugation Kit (Fast, Type A) - Lightning-Link.
Soleto, Irene, et al used Biotinylation Kit / Biotin Conjugation Kit (Fast, Type A) - Lightning-Link® (ab201795) as part of examining the effect of APRIL on B cells using rainbow trout (Oncorhynchus mykiss) as a model species. They used the kit to conjugate biotin to a recombinant trout proliferation-inducing ligand (APRIL) for use in Fluorescence microscopy.
Total leukocytes from spleen were incubated with recombinant biotinylated APRIL (1 μg/ml) for 15 min, then plated onto poly-l-lysine-coated glass slides, fixed and labeled with anti-IgM (shown as red) and FITC-streptavidin (APRIL, shown as green), then counterstained with DAPI (blue), and analyzed by confocal fluorescence microscopy.
Image from Soleto, Irene, et al., Frontiers in immunology, 9:1880. doi: 10.3389/fimmu.2018.01880; Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/
