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Np40 solution

Manufactured by Thermo Fisher Scientific
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NP40 solution is a non-ionic detergent commonly used in biological research applications. It is a versatile reagent that can be utilized for cell lysis, protein extraction, and immunoprecipitation procedures. The solution is designed to gently disrupt cell membranes while preserving the native structure and function of proteins.

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Lab products found in correlation

Qubit protein detection kit, by Thermo Fisher Scientific (4 mentions) Anti rabbit secondary antibody, by LI COR (4 mentions) Fx system, by LI COR (4 mentions) Nitrocellulose membrane, by Bio-Rad (4 mentions) N6amt1, by Santa Cruz Biotechnology (2 mentions) β tubulin, by Santa Cruz Biotechnology (2 mentions) β actin, by Santa Cruz Biotechnology (2 mentions) Blocking buffer, by LI COR (2 mentions)

Close spelling variants of this product

NP40 Detergent Surfact-Amps solution (2 citations)

4 protocols using np40 solution

1

Western Blot Protein Detection

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Protein samples were extracted by using NP40 solution followed manufactory’s protocol (Thermofisher) and protein concentration was determined by using Qubit protein detection Kit (Invitrogen) followed by manufactory’s protocol. Individual samples were run on a single 10 well gel or 12 well pre-made 4–12% gel (Thermofisher). Briefly, samples were prepared on ice (to final volume of 20 μl) and then vortexed and denatured for 10 min at 90 °C. Gels were run with 1X Tris buffered saline-Tween (TBS-T) and proteins transferred onto nitrocellulose membrane (Bio-rad). The membrane was blocked by blocking buffer (Licor) for 1hr at room temperature, washed with TBS-T for 5 min (3X), and incubated with 5ml of N6amt1 (1:250; Santa Cruz) and Beta-actin (1:500; Santa Cruz) or beta-tubulin (1:500; Santa Cruz) antibodies in blocking buffer (Licor) for overnight at 4 °C. The membranes were washed with TBS-T (3X), incubated for 1hr with anti-mouse secondary antibody (1: 15000; Li-Cor) and anti-rabbit secondary antibody (1:15000; Li-Cor) in blocking buffer (Li-Cor), and washed three times with TBST for 10 min (5X) and 20 min (1X). Optical density readings of the membrane were taken using a Li-Cor FX system followed by manufactory’s protocol. Detailed antibody information is attached within Reporting Summary file.
Li X., Zhao Q., Wei W., Lin Q., Magnan C., Emami M.R., da Silva L.E., Viola T.W., Marshall P.R., Yin J., Madugalle S.U., Wang Z., Nainar S., Vågbø C.B., Leighton L.J., Zajaczkowski E.L., Ke K., Grassi-Oliveira R., Bjørås M., Baldi P.F., Spitale R.C, & Bredy T.W. (2019). The DNA modification N6-methyl-2’-deoxyadenosine (m6dA) drives activity-induced gene expression and is required for fear extinction. Nature neuroscience, 22(4), 534-544.
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2

Western Blot Protein Detection

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Protein samples were extracted by using NP40 solution followed manufactory’s protocol (Thermofisher) and protein concentration was determined by using Qubit protein detection Kit (Invitrogen) followed by manufactory’s protocol. Individual samples were run on a single 10 well gel or 12 well pre-made 4–12% gel (Thermofisher). Briefly, samples were prepared on ice (to final volume of 20 μl) and then vortexed and denatured for 10 min at 90 °C. Gels were run with 1X Tris buffered saline-Tween (TBS-T) and proteins transferred onto nitrocellulose membrane (Bio-rad). The membrane was blocked by blocking buffer (Licor) for 1hr at room temperature, washed with TBS-T for 5 min (3X), and incubated with 5ml of N6amt1 (1:250; Santa Cruz) and Beta-actin (1:500; Santa Cruz) or beta-tubulin (1:500; Santa Cruz) antibodies in blocking buffer (Licor) for overnight at 4 °C. The membranes were washed with TBS-T (3X), incubated for 1hr with anti-mouse secondary antibody (1: 15000; Li-Cor) and anti-rabbit secondary antibody (1:15000; Li-Cor) in blocking buffer (Li-Cor), and washed three times with TBST for 10 min (5X) and 20 min (1X). Optical density readings of the membrane were taken using a Li-Cor FX system followed by manufactory’s protocol. Detailed antibody information is attached within Reporting Summary file.
Li X., Zhao Q., Wei W., Lin Q., Magnan C., Emami M.R., da Silva L.E., Viola T.W., Marshall P.R., Yin J., Madugalle S.U., Wang Z., Nainar S., Vågbø C.B., Leighton L.J., Zajaczkowski E.L., Ke K., Grassi-Oliveira R., Bjørås M., Baldi P.F., Spitale R.C, & Bredy T.W. (2019). The DNA modification N6-methyl-2’-deoxyadenosine (m6dA) drives activity-induced gene expression and is required for fear extinction. Nature neuroscience, 22(4), 534-544.
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3

Western Blot Protein Detection Protocol

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Protein samples were extracted using NP40 solution following the manufacturer’s protocol (Thermo Fisher) and the protein concentration was determined using the Qubit protein detection kit (Invitrogen), also following the manufacturer’s protocol. Individual samples were run on a single 10-well gel pre-made 4–12% gel (Thermo Fisher). Briefly, samples were prepared on ice (to a final volume of 20 μL) and then vortexed and denatured for 10 min at 90°C. Gels were run with TBS-T and proteins were transferred onto a nitrocellulose membrane (Biorad). The membrane was blocked by blocking buffer (LI-COR) for 1 h at room temperature, washed with TBS-T for 5 min (three times) and incubated with 5 mL of 14-3-3 β/α and β-tubulin (control) antibodies (Table S4) in blocking buffer (LI-COR) overnight at 4°C. The membranes were washed with TBS-T (three times), incubated for 1 h with anti-mouse secondary antibody (1:15,000; LI-COR) and anti-rabbit secondary antibody (1:15,000; LI-COR) in blocking buffer (LI-COR), then washed with TBS-T for 10 min (three times) and 20 min (once). Absorbance readings of the membrane were taken using a LI-COR FX system following the manufacturer’s protocol.
Wei W., Zhao Q., Wang Z., Liau W.S., Basic D., Ren H., Marshall P.R., Zajaczkowski E.L., Leighton L.J., Madugalle S.U., Musgrove M., Periyakaruppiah A., Shi J., Zhang J., Mattick J.S., Mercer T.R., Spitale R.C., Li X, & Bredy T.W. (2022). ADRAM is an experience-dependent long noncoding RNA that drives fear extinction through a direct interaction with the chaperone protein 14-3-3. Cell reports, 38(12), 110546.
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4

Western Blot Protein Detection Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols
Protein samples were extracted using NP40 solution following the manufacturer’s protocol (Thermo Fisher) and the protein concentration was determined using the Qubit protein detection kit (Invitrogen), also following the manufacturer’s protocol. Individual samples were run on a single 10-well gel pre-made 4–12% gel (Thermo Fisher). Briefly, samples were prepared on ice (to a final volume of 20 μL) and then vortexed and denatured for 10 min at 90°C. Gels were run with TBS-T and proteins were transferred onto a nitrocellulose membrane (Biorad). The membrane was blocked by blocking buffer (LI-COR) for 1 h at room temperature, washed with TBS-T for 5 min (three times) and incubated with 5 mL of 14-3-3 β/α and β-tubulin (control) antibodies (Table S4) in blocking buffer (LI-COR) overnight at 4°C. The membranes were washed with TBS-T (three times), incubated for 1 h with anti-mouse secondary antibody (1:15,000; LI-COR) and anti-rabbit secondary antibody (1:15,000; LI-COR) in blocking buffer (LI-COR), then washed with TBS-T for 10 min (three times) and 20 min (once). Absorbance readings of the membrane were taken using a LI-COR FX system following the manufacturer’s protocol.
Wei W., Zhao Q., Wang Z., Liau W.S., Basic D., Ren H., Marshall P.R., Zajaczkowski E.L., Leighton L.J., Madugalle S.U., Musgrove M., Periyakaruppiah A., Shi J., Zhang J., Mattick J.S., Mercer T.R., Spitale R.C., Li X, & Bredy T.W. (2022). ADRAM is an experience-dependent long noncoding RNA that drives fear extinction through a direct interaction with the chaperone protein 14-3-3. Cell reports, 38(12), 110546.
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