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Standard np 40 lysis buffer

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Standard NP-40 lysis buffer is a non-ionic detergent-based solution used for cell lysis and protein extraction. It is a commonly used reagent in various biochemical and molecular biology applications.

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Lab products found in correlation

Pierce bca protein assay kit, by Thermo Fisher Scientific (6 mentions) Westernbreeze chromogenic kit, by Thermo Fisher Scientific (3 mentions) Nbt bcip substrate, by Thermo Fisher Scientific (3 mentions) Pvdf membrane, by Merck Group (3 mentions) Anti gapdh, by Cell Signaling Technology (2 mentions) Anti phospho p65, by Cell Signaling Technology (1 mentions) Rabbit anti p65, by Cell Signaling Technology (1 mentions) Anti tbp, by Cell Signaling Technology (1 mentions) Anti gapdh 2118, by Cell Signaling Technology (1 mentions) Anti phospho pi3k, by Cell Signaling Technology (1 mentions) Anti erk, by Cell Signaling Technology (1 mentions) Anti phospho erk, by Cell Signaling Technology (1 mentions) Anti pi3k, by Cell Signaling Technology (1 mentions)

3 protocols using standard np 40 lysis buffer

1

Western Blot Analysis of Signaling Proteins

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Cell lysates were prepared in standard NP-40 lysis buffer (Abcam, USA) supplemented with proteinase and phosphatase inhibitors. Protein lysates were then quantified using the Pierce BCA protein assay kit (Thermo Fisher Scientific, USA). Equal masses of total protein were separated on 4–12% SDS-polyacrylamide mini-gels and blotted onto PVDF membrane (Millipore, USA). Membranes were subsequently blocked, incubated with primary antibodies, and incubated with secondary antibodies according to WesternBreeze Chromogenic Kit (Thermo Scientific, USA). Alkaline phosphatase was detected on the PVDF membranes using a ready-to-use BCIP/NBT substrate (Thermo Scientific, USA) for ready visualization of enzyme-linked antibodies. Rabbit anti-p65, anti-phospho-p65, anti-GAPDH, and anti-TBP antibodies were obtained from Cell Signaling (USA) or Abcam (USA).
Zgheib C., Hodges M.M., Hu J., Liechty K.W, & Xu J. (2017). Long non-coding RNA Lethe regulates hyperglycemia-induced reactive oxygen species production in macrophages. PLoS ONE, 12(5), e0177453.
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2

Western Blot Analysis of Signaling Proteins

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Cell lysates were prepared in standard NP-40 lysis buffer (Abcam, USA) supplemented with proteinase and phosphatase inhibitors. Protein lysates were quantified using the Pierce BCA protein assay kit (Thermo Fisher Scientific, USA). Equal masses of total protein were separated on 4–12% SDS-polyacrylamide mini-gels, and blotted onto PVDF membranes (Millipore, USA). Membranes were subsequently blocked, incubated with primary antibodies, and incubated with secondary antibodies according to WesternBreeze Chromogenic Kit (Thermo Scientific, USA). Alkaline phosphatase was detected on the PVDF membranes using a ready-to-use BCIP/NBT substrate (Thermo Scientific, USA) for ready visualization of enzyme-linked antibodies. Rabbit anti-STAT1(149943), anti-phospho-STAT1(76493), anti-STAT3(91393), anti-phospho-STAT3 (9131), anti-phospho-ERK(19762), and anti-GAPDH (2118) antibodies were obtained from Cell Signaling (USA). Quantification of relative intensities was achieved by ImageJ analysis.
Hu J., Zhang L., Liechty C., Zgheib C., Hodges M.M., Liechty K.W, & Xu J. (2020). Long non-coding RNA GAS5 regulates macrophage polarization and diabetic wound healing. The Journal of investigative dermatology, 140(8), 1629-1638.
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3

Western Blot Analysis of PI3K and ERK

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Cell lysates were prepared in standard NP-40 lysis buffer (Abcam, USA) supplemented with proteinase and phosphatase inhibitors. Protein lysates were quantified using the Pierce BCA protein assay kit (Thermo Fisher Scientific, Waltham, MA, USA). Equal masses of total protein were separated on 4–12% SDS-polyacrylamide mini-gels and blotted onto PVDF membranes (Millipore, Burlington, MA, USA). Membranes were subsequently blocked, incubated with primary antibodies, and incubated with secondary antibodies, according to WesternBreeze Chromogenic Kit (Thermo Scientific, Waltham, MA, USA). Alkaline phosphatase was detected on the PVDF membranes using a ready-to-use BCIP/NBT substrate (Thermo Scientific, USA) for ready visualization of enzyme-linked antibodies. Rabbit anti-PI3K, anti-phospho-PI3K, anti-ERK, anti-phospho-ERK, and anti-GAPDH antibodies were obtained from Cell Signaling (USA). Quantification of relative intensities was achieved by ImageJ analysis.
Liechty C., Hu J., Zhang L., Liechty K.W, & Xu J. (2020). Role of microRNA-21 and Its Underlying Mechanisms in Inflammatory Responses in Diabetic Wounds. International Journal of Molecular Sciences, 21(9), 3328.
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