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Dylight 800

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DyLight 800 is a fluorescent dye used for labeling and detection in various biological applications. It emits light in the near-infrared spectrum, making it suitable for use in imaging and analysis techniques that require this wavelength range.

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Lab products found in correlation

Starbright blue 700, by Bio-Rad (3 mentions) Blocking buffer, by Bio-Rad (2 mentions) Mouse anti flag, by Merck Group (2 mentions) Chemidoc imager, by Bio-Rad (2 mentions) Nupage 12 bis tris gel, by Thermo Fisher Scientific (2 mentions) Ab49763, by Abcam (2 mentions) Starbright 700, by Bio-Rad (2 mentions) Flag tag (flag), by Merck Group (1 mentions) Gapdh, by Merck Group (1 mentions) β tubulin, by Merck Group (1 mentions) C myc, by Santa Cruz Biotechnology (1 mentions) Pvdf membrane, by Bio-Rad (1 mentions) Chemidoc mp imaging system, by Bio-Rad (1 mentions) Sc 20917, by Santa Cruz Biotechnology (1 mentions) Aga 014, by Alomone (1 mentions)

6 protocols using dylight 800

1

Immunoblot Analysis of Bacterial Proteins

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TCA-treated samples were loaded onto Nu-PAGE 12% Bis-Tris gel (Invitrogen). The proteins were transferred to nitrocellulose membrane using iBlot Gel transfer block (Novex Life Technologies), blocked using Blocking Buffer (Bio-Rad), probed with polyclonal rabbit anti-RpoS (1:5000 dilution), mouse anti-Flag (Sigma, 1:2000 dilution), mouse monoclonal anti-EF-TU (LSBio, LS-C128699) (1:10,000), rabbit polyclonal anti-RssB (1:5000 dilution), and further probed with the secondary antibodies, either goat anti-mouse coupled with DyLight®800 (Bio-Rad, 1:10000 dilution) or goat anti-rabbit coupled with StarBright Blue 700 (Bio-rad, dilution 1:5000) and washed with PBST solution. The blots were visualized using fluorescent exposure with the ChemiDoc imager (Bio-Rad). Quantification was performed using Image J software (NIH).
Bouillet S., Hamdallah I., Majdalani N., Tripathi A, & Gottesman S. (2023). A negative feedback loop is critical for recovery of RpoS after stress in Escherichia coli.. bioRxiv.
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2

Blue Light Exposure Induces Tubulin Acetylation

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A 5sec on- 5sec off regime of blue light exposure was achieved using an LED-panel constructed for placement in a 37 °C, 5% CO2 incubator and controlled with an Arduino board. The blue light intensity on the cells was approx. 0.05 nW/μm2. Cells were kept under blue light for 30 minutes and then lysed in lysis buffer on ice for 20min. Lysates were loaded on SDS-PAGE gels and transferred to PVDF membranes for Western blotting. The samples were stained with monoclonal anti-AcetylTubulin antibody (Sigma: 6–11B-1,) for acetylated alpha tubulin and anti-FLAG antibody (Abcam: ab49763) for either αTAT or Z-lock αTAT at 4 C overnight. The samples were then wash and stained with dye-labeled secondary antibodies (ThermoFisher: Dylight 800; Bio-rad: Starbright 700) at room temperature for 1 hour.
Stone O.J., Pankow N., Liu B., Sharma V.P., Eddy R.J., Wang H., Putz A.T., Teets F.D., Kuhlman B., Condeelis J.S, & Hahn K.M. (2019). Optogenetic control of Cofilin and αTAT in living cells using Z-lock. Nature chemical biology, 15(12), 1183-1190.
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3

Comprehensive Antibody Panel for Cellular Signaling Analysis

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Primary antibodies included antibodies to S6K1 (Abcam 32359), pS6K-Thr389 (CST 9205), p4EBP1-Ser65 (CST 9451), 4EBP1 (CST 9644), pan AKT (CST 4691), pAKT-Thr308 (CST 2965), AKT1 (CST 2938), pAKT substrate (RXRXXpS*/T*) (CST 10001), GAPDH (CST 5174), flag (Sigma F1804), flag (Sigma A9594), HA (CST 3724), HA (CST 3444), GST (CST 2625), Ub (Santa Cruz sc-8017), c-Myc (Santa Cruz sc-40), RNF167 (Santa Cruz sc-515405), RNF167 (Proteintech 24618-1-AP), and β-tubulin (Sigma 7B9). Antibodies to CASTOR1 were described as before13 . Secondary antibodies included mouse anti-Rabbit IgG (Light-Chain Specific) (CST 93702), rabbit anti-Mouse IgG (Light Chain Specific) (CST 58802), goat anti-rabbit horseradish peroxidase (HRP)-conjugated IgG (CST 7074), horse anti-mouse IgG HRP-conjugated IgG (CST 7076), goat anti-mouse IgG DyLight 800 (Bio-Rad STAR117D800GA), and goat anti-rabbit IgG StarBright Blue700 (Bio-Rad 12004161).
Li T., Wang X., Ju E., da Silva S.R., Chen L., Zhang X., Wei S, & Gao S.J. (2021). RNF167 activates mTORC1 and promotes tumorigenesis by targeting CASTOR1 for ubiquitination and degradation. Nature Communications, 12, 1055.
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4

Quantifying Hippocampal GABA Receptor Subunits

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Dorsal hippocampi were collected around the tips of the hippocampal cannulas. Tissue was lysed in modified RIPA buffer, incubated 15 min on ice, and centrifuged for 15 min (15,000g) at 4 °C. Samples were subjected to SDS-PAGE (10 μg per well) and transferred to PVDF membrane (Biorad). Membranes were blocked with I-block (Tropix), incubated with primary antibody overnight at 4 °C, and with secondary antibody for 1 h at room temperature. Primary antibodies used were against GABAAR-α1 (1:500, Sigma-Aldrich, G4416), GABAAR-α4 (1:500, sc-20917, Santa Cruz), and GABAAR-δ (1:500, Alomone, AGA-014). Secondary antibodies were DyLight 800 (Bio Rad). Bands were visualized using ChemiDoc MP Imaging System (Bio-Rad). All antibodies gave bands at the predicted molecular sizes. Specific band intensities were normalized to the total protein bands intensity of each lane.
Jovasevic V, & Radulovic J. (2021). High ethanol preference and dissociated memory are co-occurring phenotypes associated with hippocampal GABAAR-δ receptor levels. Neurobiology of learning and memory, 183, 107459.
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5

Protein Expression Analysis Protocol

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TCA-treated samples were loaded onto Nu-PAGE 12% Bis-Tris gel (Invitrogen). The proteins were transferred to nitrocellulose membrane using iBlot Gel transfer block (Novex Life Technologies), blocked using Blocking Buffer (Bio-Rad), probed with polyclonal rabbit anti-RpoS (1:5000 dilution), mouse anti-Flag (Sigma, 1:2000 dilution), mouse monoclonal anti-EF-TU (LSBio, LS-C128699) (1:10,000), rabbit polyclonal anti-RssB (1:5000 dilution), and further probed with the secondary antibodies, either goat anti-mouse coupled with DyLight 800 (Bio-Rad, 1:10000 dilution) or goat anti-rabbit coupled with StarBright Blue 700 (Bio-rad, dilution 1:5000) and washed with PBST solution. The blots were visualized using fluorescent exposure with the ChemiDoc imager (Bio-Rad). Quantification was performed using Image J software (NIH).
Bouillet S., Hamdallah I., Majdalani N., Tripathi A, & Gottesman S. (2024). A negative feedback loop is critical for recovery of RpoS after stress in Escherichia coli. PLOS Genetics, 20(3), e1011059.
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6

Blue Light Exposure Induces Tubulin Acetylation

Check if the same lab product or an alternative is used in the 5 most similar protocols
A 5sec on- 5sec off regime of blue light exposure was achieved using an LED-panel constructed for placement in a 37 °C, 5% CO2 incubator and controlled with an Arduino board. The blue light intensity on the cells was approx. 0.05 nW/μm2. Cells were kept under blue light for 30 minutes and then lysed in lysis buffer on ice for 20min. Lysates were loaded on SDS-PAGE gels and transferred to PVDF membranes for Western blotting. The samples were stained with monoclonal anti-AcetylTubulin antibody (Sigma: 6–11B-1,) for acetylated alpha tubulin and anti-FLAG antibody (Abcam: ab49763) for either αTAT or Z-lock αTAT at 4 C overnight. The samples were then wash and stained with dye-labeled secondary antibodies (ThermoFisher: Dylight 800; Bio-rad: Starbright 700) at room temperature for 1 hour.
Stone O.J., Pankow N., Liu B., Sharma V.P., Eddy R.J., Wang H., Putz A.T., Teets F.D., Kuhlman B., Condeelis J.S, & Hahn K.M. (2019). Optogenetic control of Cofilin and αTAT in living cells using Z-lock. Nature chemical biology, 15(12), 1183-1190.
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