Round leaf discs with a diameter of 1 cm were punched from the treated tobacco leaves using a hole puncher and then stained by a GUS staining kit (Solarbio, Beijing). A total of 0.2 g treated samples was homogenized in 1.8 mL ice-cold potassium phosphate buffer (50 mM, pH 7.8) and then centrifuged at 8000 r/min for 20 min at 4 °C. The aqueous (upper) phase was analyzed by ELISA kit (Mlbio, Shanghai) for GUS activity.
Gus staining kit
Manufactured by Solarbio
Sourced in China
The GUS staining kit is a laboratory tool used to detect and visualize the activity of the β-glucuronidase (GUS) reporter gene in biological samples. The kit provides the necessary reagents and protocols to perform a histochemical staining procedure that reveals the presence and localization of the GUS enzyme within cells or tissues.
Automatically generated - may contain errors
Lab products found in correlation
Mz16f, by Leica Microsystems (2 mentions)
Elisa kit, by Mlbio (1 mentions)
M205 fa, by Leica camera (1 mentions)
Fluorescence spectrophotometer, by Thermo Fisher Scientific (1 mentions)
Mvx10, by Olympus (1 mentions)
Clonexpress 2 one step cloning kit, by Vazyme (1 mentions)
Clonexpress multis one step cloning kit, by Vazyme (1 mentions)
60d camera, by Canon (1 mentions)
11 protocols using gus staining kit
1
Quantifying GUS Activity in Tobacco Leaves
2
GUS Reporter Gene Expression in Rice
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The roots, leaves and sheaths of 10-day water-cultured rice seedlings were taken into 10-mL tubes containing 5 mL of GUS staining solution, respectively. The rice materials in GUS-dyed solution were evacuated for 15 min with a vacuum pump and then incubated at 37 °C for 24 h. Following that, the chlorophyll of rice materials was removed by ethanol, and then, the expression level of the GUS reporter gene in the rice tissue was observed. The detailed process was described in the GUS Staining Kit (No. G3060, Solarbio, Beijing, China).
3
Quantification of GUS Reporter Gene Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The β-glucuronidase (GUS) gene is a commonly used reporter gene, which could reflect promoter expression levels. GUS histochemical staining was performed on tobacco leaves using the GUS Staining Kit (Solarbio, Beijing, China). First, tobacco leaves were punched and soaked in GUS staining solution. The vacuum pump was used for 30 min under a 0.085 kPa condition, then stained overnight at 37 °C. Finally, a 75% ethanol solution was used to decolorize. GUS expression was observed and photographed under a microscope (Leica M205FA, Wetzlar, Germany). Moreover, the quantification of GUS activity was determined using the fluorometric 4-methylumbelliferyl-b-D-glucuronide (MUG) method. One unit of GUS activity was defined as 1 nM of 4-methylumbelliferon (4-MU) generated per milligram of soluble protein per minute. Three leaves were infiltrated for each construct in each independent experiment and then combined to detect GUS activity.
4
Histochemical GUS Staining Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Histochemical GUS staining was performed with GUS staining Kit, according to the manual (G3061, Solarbio Co., Beijing, China). Samples were fixed in 90% acetone at −20 °C, rinsed four times with 0.1 M sodium phosphate buffer (pH 7.4), and then incubated in X-Gluc solution [0.1 M sodium phosphate (pH 7.4), 3 mM potassium ferricyanide, 0.5 mM potassium ferrocyanide, 0.5 g L−1 5-bromo-4-chloro-3-indolyl-β-d-glucuronide cyclohexilammonium salt] at 37 °C. After staining, samples were incubated in methanol to remove chlorophyll and then mounted in the clearing solution (a mixture of chloral hydrate, water, and glycerol in a ratio of 8:2:1). Observation was performed using a stereomicroscope (MZ16F, Leica Microsystems, Germany) or a microscope equipped with Nomarski optics (BX51, Olympus Co., Tokyo, Japan).
5
GUS Activity Assay in Recombinant Apples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Apple calli carrying the recombinant plasmids were stained with a GUS Staining Kit (Solarbio, Beijing, China) according to the manufacturer’s instructions and GUS activity was determined with a fluorescence spectrophotometer (ThermoFisher Scientific).
6
GUS Staining of Transformed Calli
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The GUS staining assay for transformed calli and regenerated shoots was performed according to the GUS staining kit instructions (Solarbio, G3060, China). GUS staining buffer (2.5 mL GUS buffer A, 10 μL GUS buffer B, 10 μL GUS buffer C, 2.0 mL methanol, 20 μL X-GlcA solution, 5.5 mL deionized water) added to the samples that were vacuum infiltrated for 2 min. After incubation at 37°C for 24 h, the samples were bleached by immersion in 75% ethanol, and then observed and photographed under stereo microscope (Olympus, MVX10, Japan).
7
Tissue Expression Analysis of PmGRF6::GUS Tobacco
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The 30-day PMdGRF6::GUS transgenic tobacco was used for tissue expression analysis. Young leaf, stem, and root tissues were visualized using the GUS Staining Kit (Solarbio, Beijing, China).
8
Transient Expression of CsaV3_5G039430 in Nicotiana benthamiana
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The full-length CDS of CsaV3_5G039430 was amplified with the specific primers (Supplementary Table 1 ) and inserted into pBI121 using the ClonExpress II One Step Cloning Kit (Vazyme, Nanjing, China) to obtain 35S:: CsaV3_5G039430-GUS vector. For mutation 6 bases of miR9748 binding sites in CsaV3_5G039430, the CDS was amplified and inserted into pBI121 vector using the ClonExpress MultiS One Step Cloning Kit (Vazyme, Nanjing, China) to obtain 35S:: CsaV3_5G039430M-GUS vector. The recombinant plasmids were transformed into A. tumefaciens strain EHA105, and transiently transformed into the leaves of Nicotiana benthamiana as previously described method (Wang et al., 2020c (link)). After 2 d transformation, the leaves were stained with GUS staining kit (Solarbio, Beijing, China) and photographed.
9
GUS Staining Protocol for Plant Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Histochemical GUS staining was performed with GUS staining kit according to the manual (G3061, Solarbio, Beijing, China). Samples were fixed in 90% acetone at −20°C, rinsed four times with 0.1 m sodium phosphate buffer (pH 7.4), and then incubated in X‐Gluc solution [0.1 m sodium phosphate (pH 7.4), 3 mm potassium ferricyanide, 0.5 mm potassium ferrocyanide, 0.5 g L−1 5‐bromo‐4‐chloro‐3‐indolyl‐β‐d ‐glucuronide cyclohexilammonium salt] at 37°C. After staining, samples were incubated in methanol to remove chlorophyll and then mounted in the clearing solution (a mixture of chloral hydrate, water and glycerol in a ratio of 8:2:1). Observation was performed using a stereomicroscope (MZ16F, Leica Microsystems, Germany) or a microscope equipped with Nomarski optics (BX51, Olympus, Tokyo, Japan). For the observation of vascular patterns, cotyledons were fixed in a mixture of ethanol and acetic acid in a ratio of 9:1, hydrated through a graded series of ethanol, and then mounted with the clearing solution (Konishi and Sugiyama, 2003 (link)).
10
Characterizing WRKY Transcription Factors
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The CDS of TPR, or the promoter sequences of WRKY41 and WRKY46 were amplified using the specific primers (Supplementary Table 1 ), and inserted into the pBI121 vector. The recombinant plasmid was transformed into the A. tumefaciens strain GV3101. The tobacco was transiently transformed according to the previous method (Wang Y. et al., 2020 (link)). After transformation for 2 d, the leaves were stained by GUS staining Kit (Solarbio, Beijing, China) and photographed. For analyzing the role of ABA on the expression of WRKY41 and WRKY46, tobacco leaves were subjected to foliar spray of 100 μmol ABA after injection for 1 d.
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!