Seedlings were germinated vertically for 3 days on 1/2MS solid medium containing 0.4% Gelzan™ CM (Millipore Sigma) and then transferred to plates containing either 0 (P‐deficient) or 675 µM (P‐sufficient) phosphate. Gelzan™ (0.4% w/v) was selected as gelling agent to prevent unexpected phosphate contamination from different batches of agar (Jain et al., 2009 (link)). Three days after transferring the seedlings, RH images were captured using a Nikon SMZ1500 stereomicroscope. RH length was determined by measuring at least 700 RHs located between 2 and 6 mm from the tip of the primary root in 10 individual plants for each genotype. All the measurements were repeated at least three times with representative RH images shown in the figures.
Gelzan cm
Manufactured by Merck Group
Sourced in United States
Gelzan CM is a microbial-derived polysaccharide-based gelling agent used in various laboratory applications. It forms thermoreversible gels when dissolved in aqueous solutions. The core function of Gelzan CM is to provide a stabilizing and thickening effect in various formulations and media, without making interpretations about its intended use.
Automatically generated - may contain errors
Lab products found in correlation
Cacl2, by Merck Group (2 mentions)
Smz1500, by Nikon (1 mentions)
Cryodos 80, by Telstar (1 mentions)
Eudragit epo, by Evonik (1 mentions)
Ethylenediamine, by Thermo Fisher Scientific (1 mentions)
Ethylene glycol, by ITW Reagents (1 mentions)
Mineral oil, by Merck Group (1 mentions)
Gibco trypsin edta, by Thermo Fisher Scientific (1 mentions)
Span 85, by Merck Group (1 mentions)
Gibco penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Gibco fetal bovine serum, by Thermo Fisher Scientific (1 mentions)
Gibco dmem, by Thermo Fisher Scientific (1 mentions)
Calcium chloride, by Merck Group (1 mentions)
Fitc annexin 5 apoptosis detection kit 1, by BD (1 mentions)
Gelrite, by Merck Group (1 mentions)
P32t8 tl850, by Philips (1 mentions)
Acetonitrile, by ITW Reagents (1 mentions)
Solutol hs15, by Merck Group (1 mentions)
Calcium chloride dihydrate, by Vetec (1 mentions)
Milli q water system, by Merck Group (1 mentions)
18 protocols using gelzan cm
1
Phosphate-Induced Root Hair Growth
2
Arabidopsis Growth and Nutrient Deprivation
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Arabidopsis thaliana (ecotype Col‐0) seeds were sterilized according to Ying et al. (2012 (link)) and placed on half‐strength Murashige and Skoog (MS) medium ± phosphate (Caisson Labs, Smithfield, UT, USA), supplemented with 1% (w/v) sucrose, 0.5 mM MES‐KOH (pH 5.7) and solidified with 0.4% (w/v) Gelzan™ CM (MilliporeSigma, Burlington, MA, USA). The plated seeds were stratified for 3 d at 4°C in the dark before placing the plates vertically in a growth chamber (22°C, 120 µmol−2s−1 light intensity, 16 h : 8 h, light : dark cycle). Macronutrient deprivation treatments were done as described previously (Scheible et al., 2004 (link); Bläsing et al., 2005 (link); Morcuende et al., 2007 ; Bielecka et al., 2015 (link)). Plant materials were harvested by rinsing twice in demineralized water, gently blotting on tissue paper and snap‐freezing in liquid nitrogen (N2) before storage at −80°C until further use. The rxr1 (SALK_048882) and rabd2c (SALK_054626) T‐DNA insertion mutant lines were obtained from ABRC. Homozygous mutant plants were identified using the protocol available athttp://signal.salk.edu/ .
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Arabidopsis thaliana (ecotype Col‐0) seeds were sterilized according to Ying et al. (2012 (link)) and placed on half‐strength Murashige and Skoog (MS) medium ± phosphate (Caisson Labs, Smithfield, UT, USA), supplemented with 1% (w/v) sucrose, 0.5 mM MES‐KOH (pH 5.7) and solidified with 0.4% (w/v) Gelzan™ CM (MilliporeSigma, Burlington, MA, USA). The plated seeds were stratified for 3 d at 4°C in the dark before placing the plates vertically in a growth chamber (22°C, 120 µmol−2s−1 light intensity, 16 h : 8 h, light : dark cycle). Macronutrient deprivation treatments were done as described previously (Scheible et al., 2004 (link); Bläsing et al., 2005 (link); Morcuende et al., 2007 ; Bielecka et al., 2015 (link)). Plant materials were harvested by rinsing twice in demineralized water, gently blotting on tissue paper and snap‐freezing in liquid nitrogen (N2) before storage at −80°C until further use. The rxr1 (SALK_048882) and rabd2c (SALK_054626) T‐DNA insertion mutant lines were obtained from ABRC. Homozygous mutant plants were identified using the protocol available at
3
Fabrication of Lactoferrin-Gellan Hydrogels
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Gelzan CM (Sigma, St. Louis, MO, USA) was dissolved in distilled water at 90 °C for 30 min. with stirring. Lactoferrin (Lf, Bovine origin, New Zealand) was dissolved in distilled water. Subsequently, to the solution of Gellan Gum (GG), Lactoferrin (Lf), and Hydroxyapatite (HAp, Plasma Biotal, Buxton, UK) were added at 50 °C, as described in Table 1 . After complete homogenization, the crosslinking solution CaCl2 (Sigma, USA) was added and it was allowed to stabilize at room temperature for 30 min. Posteriorly, the hydrogels were cut in discs of 5 mm of diameter and 2 mm of thickness and replenished with phosphate-buffered saline solution (PBS, Sigma, USA) for 30 h to enable complete crosslinking, frozen at −80 °C for 18–20 h, and then freeze-dried (CryoDos -80, Telstar, Terrassa, Barcelona, Spain) for at least three days. Materials were sterilized by ethylene oxide.
4
Curcumin and Resveratrol Formulation Development
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Curcuminoid extract powder derived from Curcumin longa (96.1% curcumin) was purchased from Thai-China Flavours and Fragrances Industry Co., Ltd. (Phra Nakhon Si Ayutthaya, Thailand). Resveratrol (3,5,4′-trihydroxy-trans-stilbene; purity > 98%) was purchased from Pioneer Herb (Shanghai, China). Eudragit® EPO was provided by Evonik Industries AG (Essen, Germany). Sodium alginate (medium viscosity) was obtained from High Science Limited Partnership (Songkhla, Thailand). Sodium alginate (low viscosity) and gellan gum (Gelzan™ CM) were purchased from Sigma-Aldrich, Inc. (St. Louis, MO, USA). Pectin (GENU®), low methoxy grade, was sourced from CP Kelco, Inc. (Atlanta, GA, USA). Sodium bicarbonate and ethanol were provided by RCI Labscan (Bangkok, Thailand). Calcium carbonate was obtained from Vidhayasom Co., Ltd. (Bangkok, Thailand). Polyvinylpyrrolidone (PVP K30) was purchased from P.C. Drug Center Co., Ltd. (Bangkok, Thailand). All reagents were of pharmaceutical or analytical grade.
5
Synthesis and Characterization of Doxorubicin-Loaded Nanoparticles
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Gelzan™ CM, calcium chloride, mineral oil, doxorubicin hydrochloride (Dox-HCl) and Span® 85 were purchased from Sigma-Aldrich (St. Louis, MO, USA). Gibco™ DMEM, Gib-co™ Fetal Bovine Serum (FBS), Gibco™ Penicillin-Streptomycin, Invitrogen™ Presto-Blue™ Cell Viability Reagent and Gibco™ Trypsin-EDTA (0.05%) were purchased from Thermo Fisher Scientific (Branchburg, NJ, USA). FITC Annexin V Apoptosis Detection Kit I was purchased from BD Pharmingen™ (San Diego, CA, USA). Sodium Nitrate, hydrogen peroxide and Sodium acetate were purchased from HAYASHI PURE CHEMICAL IND. (Osaka, Japan), Ltd. (HPC). Sulfuric Acid was purchased from UNION CHEMICAL WORKS Ltd. (Hsinchu, Taiwan). Potassium permanganate and ferric chloride were purchased from SHIMAKYU’S PURE CHEMICALS (Osaka, Japan). Ethylenediamine was purchased from ACROS Organics™ (Carlsbad, CA, USA). Ethylene glycol was purchased from PanReac AppliChem (Chicago, IL, USA). Graphite powder was donated from the Department of Public Health (Taichung, Taiwan), Chung Shan Medical University. All the samples in this study were sterilized in 70% (v/v) ethanol solution at 25 °C for 24 h. Next, samples were washed three times in sterile phosphate buffered saline (PBS) for five minutes each time. Finally, samples were dried or used directly for testing.
6
Tomato Micro-Tom Seed Transformation
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Tomato Micro-Tom seeds (Moles Seeds, cat. # VTO325) were surface sterilized, rinsed and soaked for 24 h in smoke-water (0.5% v/v) [39 ], and then rinsed for 1 min in three changes of sterile distilled water. Next, seeds were manually scarified, and then placed on MS basal medium (Sigma-Aldrich cat. # M5519) [40 ] supplemented with 3g/L activated charcoal plus 3 g/L Gelrite (Sigma-Aldrich, Gelzan CM, Gelrite cat. # G1910) and incubated in a Percival growth chamber (Percival AR-36, Percival, Perry, IA, USA) at 22°C under long day conditions (16 h light/8 h darkness, with an irradiance of 50 μmol/m−2 s−1, using fluorescent T8 Phillips P32T8 /TL850 lamps). After 8 days, the seedlings were dissected using a stereo microscope as follows: the cotyledons were removed from the embryonic axis using a dissecting scalpel and 3 mm long cotyledon explants were used for transformation.
7
In Vitro Culture of Plants and Roots
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The plants and roots were cultured and kept in glass bottles with 50 mL of medium semi-solid as described by Moreno-Anzúrez et al. [3 (link)]. The culture medium was prepared with the macro- and micro-nutrients of MS [31 (link)] medium and the vitamins in Gamborg B5 [32 (link)] medium (MS/B5 medium). It was supplemented with 100 mg/L myo-inositol; 30, 20, 10 or 5 g/L sucrose; and 3 g/L Gelzan™ CM (Sigma-Aldrich) as a gelling agent. The same MS/B5 medium without sucrose was used for seed germination. The pH was adjusted to 5.7 ± 0.1 with potassium hydroxide, and the medium was autoclaved at 108 kPa and 121 °C for 20 min. All the cultures were incubated at a constant temperature (25 ± 2 °C) under a 16/8 h light/dark photoperiod using a 27 µmol·m2·s−1 white light illumination intensity. Each subculture lasted for 30 days. The data obtained were analysed by Chi square test using VassarStats program (VassarStats: Statistical Computation Website).
8
Curcumin and Quercetin Topical Formulation
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Curcumin (CUR), quercetin (QU), cetalkonium chloride, deacetylated gellan gum (Gelzan™ CM), polyethylene glycol stearate 660 (Solutol HS15®), castor oil, and porcine stomach mucin (type II) were bought from Sigma–Aldrich (St. Louis, MO, USA). Egg phospholipids with 80% phosphatidylcholine (Phospholipon E80®) and purified fish oil (DHA/EPA) were obtained from Lipoid (Ludwigshafen, Germany). Acetonitrile of high-performance liquid chromatography (HPLC) grade was purchased from Panreac (Barcelona, Spain), and the pure water was achieved with a Milli-Q® water system (Millipore, Burlington, MA, USA). Polyethylene glycol 400 (PEG400), potassium chloride, and sodium chloride were obtained from Synth (São Paulo, SP, Brazil). Monobasic sodium phosphate and dibasic sodium phosphate were purchased from Alphatec (Porto Alegre, RS, Brazil). The calcium chloride dihydrate was purchased from Vetec (Rio de Janeiro, RJ, Brazil). Ethanol, acetone, phosphoric acid, and other reagents were of analytical grade.
9
Quantifying Root Hair Growth Dynamics
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Seeds were surface-sterilized in ethanol and bleach as described above. To evaluate root hair growth rate, seeds were planted on 48 × 64 mm coverslips coated with 0.5× MS, 1% sucrose, and 0.4% (w/v) Gelzan CM (Sigma Aldrich, USA) according to Dyachok et al. (2016) . The coverslips were placed in 9-cm round Petri dishes and the seeds stratified at 4°C for 48 h. After stratification, the coverslip system with planted seed were kept in a 24°C growth chamber under a 14-h light (120 µmol m−2s−1)/10-h dark cycle for 5–6 d.
To quantify root hair growth rate, time-lapse sequences of elongating root hairs at intervals of 10 min over a period of 60 min from a region of the primary root located between 2 and 3 mm from the root tip were captured with a Nikon Eclipse TE300 inverted microscope using a 10× objective. Root hair lengths at various time points were extracted from digital images using ImageJ (v1.51) software (https://imagej.nih.gov/ij/ ). The displacement of the root hair tip after each 10-min interval was obtained and divided by 10 to obtain growth rate as µm/min.
To quantify root hair growth rate, time-lapse sequences of elongating root hairs at intervals of 10 min over a period of 60 min from a region of the primary root located between 2 and 3 mm from the root tip were captured with a Nikon Eclipse TE300 inverted microscope using a 10× objective. Root hair lengths at various time points were extracted from digital images using ImageJ (v1.51) software (
10
Fabrication of GG/SS Hydrogel Composites
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Four
hydrogel solutions of different GG/SS weight ratios were prepared.
Briefly, 100 mg of low-acyl GG (GG; Gelzan CM, Sigma-Aldrich, USA)
was dissolved in 9 mL of distilled water under constant stirring for
1 h at 90 °C. The temperature was then lowered to 70 °C,
and 0.01% (w/v) of calcium chloride (CaCl2, Sigma-Aldrich,
USA) as a cross-linking agent was homogeneously mixed in the GG solution
for 20 min. The different amounts of SS powder (0.5, 1, and 5%) were
completely dissolved in 1 mL of distilled water at 70 °C. The
prepared SS solution was added in the GG solution to fabricate total
1% (w/v) GG with 0, 0.05, 0.1, and 0.5% (w/v) SS, and the blended
solution was stirred for 20 min. The samples were specified as GG/SS
0%, GG/SS 0.05%, GG/SS 0.1%, and GG/SS 0.5%, respectively. The sample
named GG/SS 0% was added with distilled water instead of the SS solution.
7 mL of the prepared hydrogel solutions was poured into Petri dishes
(50 mm × 10 mm, SPL Life Sciences Co., Ltd., South Korea) and
solidified at room temperature for 10 min. After gelation was completed,
cylindrical-shaped hydrogels with a diameter of 6 mm and a height
of approximately 3 mm were prepared using a biopsy punch (Kai Medical
Biopsy Punch, Japan).
hydrogel solutions of different GG/SS weight ratios were prepared.
Briefly, 100 mg of low-acyl GG (GG; Gelzan CM, Sigma-Aldrich, USA)
was dissolved in 9 mL of distilled water under constant stirring for
1 h at 90 °C. The temperature was then lowered to 70 °C,
and 0.01% (w/v) of calcium chloride (CaCl2, Sigma-Aldrich,
USA) as a cross-linking agent was homogeneously mixed in the GG solution
for 20 min. The different amounts of SS powder (0.5, 1, and 5%) were
completely dissolved in 1 mL of distilled water at 70 °C. The
prepared SS solution was added in the GG solution to fabricate total
1% (w/v) GG with 0, 0.05, 0.1, and 0.5% (w/v) SS, and the blended
solution was stirred for 20 min. The samples were specified as GG/SS
0%, GG/SS 0.05%, GG/SS 0.1%, and GG/SS 0.5%, respectively. The sample
named GG/SS 0% was added with distilled water instead of the SS solution.
7 mL of the prepared hydrogel solutions was poured into Petri dishes
(50 mm × 10 mm, SPL Life Sciences Co., Ltd., South Korea) and
solidified at room temperature for 10 min. After gelation was completed,
cylindrical-shaped hydrogels with a diameter of 6 mm and a height
of approximately 3 mm were prepared using a biopsy punch (Kai Medical
Biopsy Punch, Japan).
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