Anti IgG (H&L)  
DyLight (商標名)

Western blot using Donkey Anti-Mouse IgG DyLight？680. TGF‐β‐mediated crosstalk between pericytes and CRC cells modulates pericyte secretome. (A) Incubation in HCT116 CM for 1？h induces SMAD3 phosphorylation in PC, as assessed by western blot. Exogenous recombinant TGF‐β (10？ng・mL−1) was used as a positive control, and β‐actin was used as loading control (n？=？3). (C) Western blot showing increased expression of αSMA in PC cocultured with HCT116 cells or stimulated with 10 ng・mL−1 TGF‐β1 for 48 h (n = 3). α‐tubulin was used as loading control. Numbers indicate the expression fold change relative to the loading control. Fig. 5. PMID: 32767843.

Western blot using Donkey Anti-Mouse IgG DyLight？680.Insulin‐like growth factor‐binding protein 3 increases CRC cell migration and invasion through Akt activation. (D) Treatment with 50？ng・mL−1？IGFBP‐3 for 72？h promotes the expression of N‐cadherin in HCT116 cells as assessed by western blot (top panel). Phosphorylation status of Akt (middle panel) and MAPK (bottom panel) in HCT116 cells treated with 50？ng・mL−1？IGFBP‐3 for 15？min. Representative images of three independent experiments (n？=？3). Numbers indicate the expression fold change relative to the loading control. Fig. 7. PMID: 32767843.

Western blot using Donkey Anti-Mouse IgG DyLight？680.The HRSV M protein co-immunoprecipitates with the AP-3Mu3A and AP-3delta complex during HRSV infection. HEp2 cells at approximately 90% confluency were either infected at an MOI of 5 or mock infected for 24 hours, cells were scraped or proteins were subsequently extracted using MPER. Cell lysates were incubated for 6 hours with 1 μg of either polyclonal goat anti-AP-3Mu3A or monoclonal mouse anti-AP-3delta along with a antibody specific isotype control, polyclonal goat-anti H1N1or monoclonal mouse anti-HIV-1 p24 Gag at 4°C on a rotating device. 20μl Protein A/G agarose beads were added to lysate plus corresponding antibody and incubated overnight. Immunoprecipate complex was pelleted and washed with PBS and then ran out on a SDS-PAGE gel and transferred to nitrocellulose membrane. Membrane was blocked and then probed with either monoclonal mouse anti-Matrix or polyclonal goat-anti HRSV primary antibody as described previously for one hour. Membranes were then washed with a PBS-Tween20 solution extensively and then probed with species-specific secondary antibodies donkey anti-goat DyLight800 and donkey anti-mouse DyLight680. Membranes were again washed extensively and blots were imaged on Odyssey Infrared imager. The results were reproducible in at least two independent assays. Fig 2. PMID: 29028839.

Western blot using Donkey Anti-Mouse IgG DyLight？680. Kif2a is enriched on spindles in？H. boettgeri？egg extracts, and inhibition of kif2a increases spindle length. B. Top Right Panel: Western blot of？X. laevis, X. tropicalis, and？H. boettgeri？extracts, probed for kif2a. Bottom Panel: Quantification of 3 separate blots for each species. Band intensities were normalized to the integrated density of the corresponding Ran loading control. AU=arbitrary units. Figure 3. PMID: 31630945.

Serine 252 of kif2a regulates its activity.B. Top Panel: Increasing amounts of recombinant？X. laevis？or？H. boettgeri？kif2a proteins were added to taxol-stabilized microtubules and microtubules sedimented through a sucrose cushion. Amounts of soluble tubulin in the supernatant (S) and pellet (P) were quantified by SDS-PAGE and Coomassie staining. Bottom Panel: Ratio of pellet to supernatant gel band intensities in the microtubule sedimentation assay with 200 nM？H. boettgeri？or？X. laevis？kif2a added. Bands from 3 separate gels quantified, p=0.2768. NS= Not Significant. Error bars= +/− std dev. Figure 4. PMID: 31630945.

Properties of DyLight？ Conjugates.

DyLight？ dyes can be used for two-color western blot detection with low background and high signal. Anti-tubulin was detected using a DyLight？ 680 conjugate. Anti-TNFa was detected using a DyLight？ 800 conjugate. The image was captured using the OdysseyR Infrared Imaging System developed by LI-COR.