MAPK activation was detected following the described procedure [35 (link),71 (link)]. Briefly, rice seedlings were treated with chitin (10 μg mL−1) or sterile H2O with 0.01% Silwet L-77 (GE Healthcare). The treated seedlings were collected for protein extraction at 15 min after treatment. Total protein extracts were subjected to immunoblotting with anti-phospho-p44/42 MAPK antibody (Cell Signaling Technology, Danvers, MA, USA).
Silwet l 77
Manufactured by GE Healthcare
Sourced in China, United States, United Kingdom, Sweden
Silwet L-77 is a surfactant used in various laboratory applications. It is a non-ionic, organo-modified siloxane that can enhance wetting and spreading properties of liquids. The product's core function is to improve the surface activity and penetration of solutions.
Automatically generated - may contain errors
Lab products found in correlation
Anti phospho p44 42 mapk antibody, by Cell Signaling Technology (2 mentions)
Silicone thinner, by Smooth-On (1 mentions)
Dhr 1, by TA Instruments (1 mentions)
Ecoflex 00 30, by Smooth-On (1 mentions)
Reverse transcription system, by Takara Bio (1 mentions)
Anti rabbit hrp conjugated secondary antibody, by CWBIO (1 mentions)
Sybr primescript rt pcr kit, by Takara Bio (1 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
Cell strainer, by BD (1 mentions)
Anti flag m2 magnetic bead, by Merck Group (1 mentions)
Sds page gel, by Bio-Rad (1 mentions)
7 protocols using silwet l 77
1
MAPK Activation in Rice Seedlings
2
Kinetin Application in Arabidopsis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For kinetin’s application, it was dissolved in 0.1 N sodium hydroxide (NaOH), to make a stock solution. The leaves of 4-weeks-old Arabidopsis plants were sprayed with a solution of 0.015% (vol/vol) Silwet L-77 (GE Healthcare, Beijing, China) containing the indicated concentrations of kinetin (Biosharp, Shanghai, China) for 3 days, three times per day. The plants were covered with transparent hoods immediately after their spraying, to retain the humidity. Mock treatments were sprayed with a solution containing only 0.015% (vol/vol) Silwet L-77.
3
Fabrication of Stretchable Microfluidic Devices
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Room temperature liquid alloy galinstan, GaInSn (68.5 wt% gallium, 21.5 wt% indium, 10 wt% tin, with viscosity of 2.4 × 10−3 Pa∙s, from Geratherm Medical AG, Geschwenda, Germany) was used as the inner flow. It was used without any pre-processing and post-processing [30 (link),31 (link)].
A silicone (polydimethylsiloxane, PDMS) kit, Elastosil RT601 A and B (mixed at a weight ratio of 9:1, Wacker Chemie, Munchen, Germany) was used as outer channel flow. The diluted PDMS was prepared by adding 3 wt% thinner (Silicone Thinner, Smooth-On, Macungie, PA, USA) to the mixed PDMS. To tune the surface tension, a 0.33 wt% surfactant Silwet L-77 (GE, Boston, MA, USA) was added to the PDMS mixture. Afterwards, the whole mixtures were put in vacuum for 10 min to eliminate bubbles. Ecoflex 0030 (Smooth-On, Macungie, PA, USA), another kind of silicone kit (mixed at a weight ratio of 1:1), was used to improve the stretchability of printed devices.
The viscosities of pure PDMS, diluted PDMS and surfactant added PDMS are 4.73 Pa∙s, 4.39 Pa∙s and 13.2 Pa∙s, respectively, which are measured by a rheometer (DHR-1, TA, New Castle, DE, USA) after the silicones were prepared for 10 min. All measurements were conducted at 20 °C with preliminary equilibration time of 1 min.
A silicone (polydimethylsiloxane, PDMS) kit, Elastosil RT601 A and B (mixed at a weight ratio of 9:1, Wacker Chemie, Munchen, Germany) was used as outer channel flow. The diluted PDMS was prepared by adding 3 wt% thinner (Silicone Thinner, Smooth-On, Macungie, PA, USA) to the mixed PDMS. To tune the surface tension, a 0.33 wt% surfactant Silwet L-77 (GE, Boston, MA, USA) was added to the PDMS mixture. Afterwards, the whole mixtures were put in vacuum for 10 min to eliminate bubbles. Ecoflex 0030 (Smooth-On, Macungie, PA, USA), another kind of silicone kit (mixed at a weight ratio of 1:1), was used to improve the stretchability of printed devices.
The viscosities of pure PDMS, diluted PDMS and surfactant added PDMS are 4.73 Pa∙s, 4.39 Pa∙s and 13.2 Pa∙s, respectively, which are measured by a rheometer (DHR-1, TA, New Castle, DE, USA) after the silicones were prepared for 10 min. All measurements were conducted at 20 °C with preliminary equilibration time of 1 min.
4
Agrobacterium-mediated Transformation of Arabidopsis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
In this study the wild type control is Arabidopsis ecotype Columbia-0 (Col-0) and soybean Zhongdou32. The BR-deficient mutant bri1-5 in WS ecotype was transformed for the BR signaling rescue experiment. The tobacco Nicotiana benthamiana was used for BR induced nuclear localization and dephosphorylation of GmBZL2. All the plants were grown in the greenhouse where the temperature of 22 °C under 16 hr light and 8 hr dark.
Agrobacterium-mediated transformation was performed with floral dip method. The positive clones were cultured in YEP medium (peptone 10 g/L, yeast extract 10 g/L, NaCl 5 g/L, pH 7.2) with 50 mg/L rifampicin and 50 mg/L kanamycin for 10 hr at 28 °C, and the Agrobacterium was collected and diluted with 5% sucrose solution containing 0.02% (v/v) Silwet L-77 (SL77080596, GE) to OD600 between 0.8–1.0. The Arabidopsis inflorescences were dipped into the mixture buffer, sealed and kept in dark overnight.
Agrobacterium-mediated transformation was performed with floral dip method. The positive clones were cultured in YEP medium (peptone 10 g/L, yeast extract 10 g/L, NaCl 5 g/L, pH 7.2) with 50 mg/L rifampicin and 50 mg/L kanamycin for 10 hr at 28 °C, and the Agrobacterium was collected and diluted with 5% sucrose solution containing 0.02% (v/v) Silwet L-77 (SL77080596, GE) to OD600 between 0.8–1.0. The Arabidopsis inflorescences were dipped into the mixture buffer, sealed and kept in dark overnight.
5
Rice Seedling Defense Responses to Elicitors
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Seven‐day‐old rice seedlings were pretreated with 10 μM DEX and mock solution overnight, and were then treated with 1 µM flg22, 10 µg/mL chitin or sterile H2O plus 0.01% Silwet L‐77 (GE Healthcare, Amersham, UK). The seedlings were collected at 6 h after treatment for detection of defence‐marker gene expression and at 15 min for MAPK activation assay. Total RNAs were isolated using TRIzol reagent (Invitrogen, Carlsbad, CA, USA) from the leaves of treated seedlings following the manufacturer’s instructions. The synthesis of cDNA was performed with the Reverse Transcription System (Takara, Dalian, China) using oligo‐d(T)18 as primers. The transcript levels of defence‐marker genes were analysed using the SYBR PrimeScript RT‐PCR kit (Takara) on an ABI PRISM 7500 system and normalized to the reference gene OsActin. The proteins were isolated from rice seedlings and MAPK activation was detected as described (Wang et al., 2015 ). Phospho‐serine/‐threonine was detected using anti‐Phospho‐p44/42 MAPK antibody (1:2000 dilution) (Cell Signaling Technology, Danvers, MA, USA) in 4 °C overnight, followed by anti‐rabbit‐HRP conjugated secondary antibody (CWBio) at 1:5000 dilution.
6
Arabidopsis Transformation via Agrobacterium
Check if the same lab product or an alternative is used in the 5 most similar protocols
Agrobacterium tumefaciens carrying the empty vector pER8 or pER8::3×Flag‐PpE4 was cultured and suspended in a solution of 5% sucrose and 0.02% Silwet L‐77 (GE Healthcare, Uppsala, Sweden). Arabidopsis ecotype Col‐0 was transformed by dipping in the suspension as described previously (Clough and Bent,1998 ). The kanamycin‐resistant seedlings were screened on selective medium and planted in soil. Then, the expression level of PpE4 in transgenic plants after induction by 17‐β‐estradiol was determined by semi‐quantitative PCR (Zuo et al., 2000 ).
+ Expand
Agrobacterium tumefaciens carrying the empty vector pER8 or pER8::3×Flag‐PpE4 was cultured and suspended in a solution of 5% sucrose and 0.02% Silwet L‐77 (GE Healthcare, Uppsala, Sweden). Arabidopsis ecotype Col‐0 was transformed by dipping in the suspension as described previously (Clough and Bent,
7
ABA-induced PYL1 protein complex
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Full-length PYL1 cDNA was subcloned into binary vector pGWB617, and cDNAs of AEL1-4 were subcloned into pGWB611 by Gateway LR recombination reaction (Invitrogen). Resultant constructs were transformed into Agrobacterium strain GV3101, which was infiltrated together with p19 strain in N. benthamiana. After 2 days, the infiltrated leaves were sprayed with water containing 0.01% Silwet L-77 (GE Healthcare) and either 0.08% ethanol or 100 mM ABA (in 0.08% ethanol) for 24 h. The harvested infiltrated leaves ($1.0 g) were ground into powder in liquid nitrogen and proteins were extracted using extraction buffer (20 mM Tris-HCl [pH 7.5], 150 mM NaCl, 0.1% NP-40, 1 mM DTT) containing a protease inhibitor cocktail ''Complete'' and phosphatase inhibitor cocktail ''PhosSTOP''. Supernatant was filtrated with a Cell Strainer (BD Falcon, 352350) and used for immunoprecipitation as an Input. The Input was incubated with 40 ml of Anti-FLAG M2 Magnetic Beads (M8823, Sigma) for 3 h (4 C), then the beads were washed and eluted according to the manufacturer's instructions. Protein samples were separated by 10% SDS-PAGE gel (Bio-Rad) and analyzed by western blotting.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!