Expanded SEJVECs on group II (1% SDS) BA scaffold were fixed in 4% paraformaldehyde (Himedia, Mumbai, India) and stained by FITC-conjugate Lycopersicon esculentum+/+ stain (lectin positive) prepared 2 μg/mL (1 h) (Zeiss, Apotome fluorescent microscope) (Favre et al. 2003 (link)).
Paraformaldehyde (pfa)
Manufactured by HiMedia
Sourced in India
Paraformaldehyde is a white, solid chemical compound that is commonly used as a fixative and preservative in various laboratory applications. It is the solid polymer of formaldehyde and is soluble in water and other organic solvents. Paraformaldehyde is a versatile reagent that can be used to preserve and maintain the structural integrity of biological samples for further analysis and experimentation.
Automatically generated - may contain errors
Lab products found in correlation
Ethidium bromide, by HiMedia (4 mentions)
Prolong gold antifade reagent, by Thermo Fisher Scientific (3 mentions)
Triton x 100, by Merck Group (3 mentions)
Trypsin edta, by Thermo Fisher Scientific (3 mentions)
Hoechst, by Merck Group (3 mentions)
Ammonium persulfate, by HiMedia (3 mentions)
Coverglass for cell growth, by Thermo Fisher Scientific (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Dynasore, by Merck Group (2 mentions)
Dimethyl sulfoxide (dmso), by HiMedia (2 mentions)
Moviol, by Merck Group (2 mentions)
Lactose, by Merck Group (2 mentions)
Rat tail collagen 1, by Thermo Fisher Scientific (2 mentions)
Phosphate buffered saline (pbs), by HiMedia (2 mentions)
Triton x 100, by HiMedia (2 mentions)
Methanol, by HiMedia (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Mowiol, by Merck Group (2 mentions)
50 bp dna ladder, by Merck Group (2 mentions)
Transferrin a488, by Merck Group (2 mentions)
18 protocols using paraformaldehyde (pfa)
1
Lycopersicon esculentum Staining of SDS-BA Scaffold
2
Imaging Cytotoxic Compound Effects on HeLa Cells
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HeLa cells (60,000) were grown onto a coverslip (Coverglass for cell Growth™, Fisherbrand, USA) in a 24-well plate. The cells were treated with 6e , 6j , and 6n compounds at their IC50 dose for 12 h, 14 h, 16 h, and 18 h. Thereafter, cells were fixed in 4% paraformaldehyde (Himedia), washed with ice-cold 1X PBS, permeabilized with 0.25% Triton X-100 (Sigma), and washed with 0.1% PBST. The cell was counter stained with 49,6-diamidino-2-phenylindole, dihydrochloride (DAPI, Invitrogen) and mounted using one drop of ProLong® Gold antifade reagent (Invitrogen) on a slide. Images were acquired by fluorescence microscope (Leica SP8 TCS) and analyzed using ImageJ (NIH).
3
Fixation and Preservation Protocol
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Paraformaldehyde, Sodium Phosphate Buffer (pH 7.4), and all other chemicals used in this study were purchased from HiMedia Laboratories, India.
4
Brain Tissue Processing for Histology
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The experimental mice were deeply anesthetized and transcardially perfused with 0.9% sterile saline followed by 4% paraformaldehyde (PFA) (Himedia, India). The brains were dissected from the skulls of animals and soaked in 4% PFA for 24 hours (h). Then the brains were transferred into separate sterile tubes containing 30% sucrose (SRL, India) and maintained at 4 °C. After a week, each brain was molded in a solution of optimal cutting temperature compound (OCT) (Sigma Aldrich, St. Louis, MO, USA) and placed on the tissue holder of a sliding microtome (Weswox, India) and the brain was frozen using dry ice. Each brain was cut into 30 µm serial sagittal sections and systematically collected in 12 sterile tubes containing a mixer of cryoprotectant solution made up of glycerol (Merck, Germany), ethylene glycol (Himedia, India), and a phosphate buffer in a 1:1:2 ratio and stored at −20 °C. The brain sections, 1 out of 12 tubes (360 µm apart) were used for histological and immunohistochemical assessments [19 (link),37 (link)].
5
Synthesis and Characterization of Biomaterials
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Para-phenylenediamine (PPDA), Moviol, Hoechst, dynasore, galectin3 (Gal3) (alexa 647), transferrin (Tf)-A488, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) were obtained from Sigma-Aldrich. Diphenyl ether (99%) was obtained from Avra, and n-hexane of HPLC grade, acetone (>99.5%), N,N-dimethyl formamide (>99.5%), ethanol(>99.5%), methanol (>99.8%), acetone, acetonitrile, isopropyl alcohol (IPA) and lactose were purchased from Merck. Paraformaldehyde, dimethyl sulfoxide (DMSO), rhodamine B, and cell culture dishes for adherent cells (treated surface) were procured from Himedia. Ham's Nutrient Mixture F12 (HAMs F12), Dulbecco's modified Eagle's medium (DMEM), fetal bovine serum (FBS), penicillin–streptomycin, trypsin–EDTA (0.25%), and collagen1 rat tail were purchased from Gibco. All the chemicals were of analytical grade and no further purification was required.
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Para-phenylenediamine (PPDA), Moviol, Hoechst, dynasore, galectin3 (Gal3) (alexa 647), transferrin (Tf)-A488, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) were obtained from Sigma-Aldrich. Diphenyl ether (99%) was obtained from Avra, and n-hexane of HPLC grade, acetone (>99.5%), N,N-dimethyl formamide (>99.5%), ethanol(>99.5%), methanol (>99.8%), acetone, acetonitrile, isopropyl alcohol (IPA) and lactose were purchased from Merck. Paraformaldehyde, dimethyl sulfoxide (DMSO), rhodamine B, and cell culture dishes for adherent cells (treated surface) were procured from Himedia. Ham's Nutrient Mixture F12 (HAMs F12), Dulbecco's modified Eagle's medium (DMEM), fetal bovine serum (FBS), penicillin–streptomycin, trypsin–EDTA (0.25%), and collagen1 rat tail were purchased from Gibco. All the chemicals were of analytical grade and no further purification was required.
6
Neurochemical Assessment of Cuprizone-Induced Demyelination
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Diazepam, mitoxantrone HCl, and quetiapine fumarate were obtained as gift sample from Intas Pharmaceuticals, Ahmedabad, India. Sodium valproate was obtained as a gift sample from Chemclone Industries, Ahmedabad, India. Phenobarbitone sodium was purchased from Abbott India Ltd., Mumbai, India. Sodium chloride, potassium chloride, glucose, disodium hydrogen phosphate, potassium dihydrogen phosphate, hydrochloric acid, and glacial acetic acid were obtained from Qualigen Fine Chemicals, Mumbai, India. All the reagents and chemicals used for the study were of analytical grade. Paraformaldehyde, percoll, sucrose, disodium EDTA, Tris buffer, HEPES sodium salt, sodium bicarbonate, calcium chloride, magnesium chloride, 4-aminopyridine, glutamate, NAD+ (oxidized form), cresyl violet acetate stain, and luxol fast blue stain were purchased from Himedia Laboratories Pvt. Ltd., Mumbai, India. Cuprizone, Complete Freund's Adjuvant (CFA), and L-glutamic acid dehydrogenase solution (bovine liver origin) were obtained from Sigma Aldrich, USA. Isoflurane (Raman and Weil Pvt. Ltd., Daman, India) was used for anesthetizing animals. Heparin was purchased from Biological E. Ltd, Hyderabad, India. Deionised water for HPLC was prepared in-house using a Milli-Q integral water purification system (Millipore Elix, Germany).
7
Antibodies and Cell Lines for Cancer Research
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Primary antibodies
such as PRDX2 (LFMA0144, Invitrogen), CHEK2 (MA515416,
Invitrogen), 53BP1 (PA116565, Invitrogen), GAPDH (ITT5052, Immunotag),
γ-H2AX (PA528778, Invitrogen), and cleaved caspase-3
(PA516335, Invitrogen) were used in this study. We procured and used
the following secondary antibodies: (i) goat anti-rabbit IGG-HRP-conjugated
secondary antibody (656120, Invitrogen), (ii) anti-rabbit IgG (H +
L), F(ab′)2 fragment (Alexa Fluor 555 Conjugate) (4413, Cell
Signaling technology), and (iii) goat anti-rabbit IgG (H + L) cross-adsorbed
secondary antibody, Alexa Fluor 488 (A-11008, Invitrogen). siRNAs
for PRDX2 and GAPDH were purchased from Ambion (Invitrogen, Thermo
Scientific). Lipofectamine was purchased from Invitrogen (Thermo Scientific).
An Amplex red hydrogen peroxide/peroxidase assay kit was purchased
from Invitrogen (Thermo Scientific). MTT (with 98% purity), fetal
bovine serum (FBS), trypsin (with 98% purity), PBS, glycerol (with
98.2% purity), Triton X-100 (99%), Bradford reagent, and paraformaldehyde
(with 99% purity) were obtained from Hi-Media Labs. CHEK2-proficient HCT116 human colorectal cancer cells were purchased from
NCCS, Pune. CHEK2-knockout HCT116 cells were given
as a kind gift by Dr. Bert Vogelstein from John Hopkins University.
such as PRDX2 (LFMA0144, Invitrogen), CHEK2 (MA515416,
Invitrogen), 53BP1 (PA116565, Invitrogen), GAPDH (ITT5052, Immunotag),
γ-H2AX (PA528778, Invitrogen), and cleaved caspase-3
(PA516335, Invitrogen) were used in this study. We procured and used
the following secondary antibodies: (i) goat anti-rabbit IGG-HRP-conjugated
secondary antibody (656120, Invitrogen), (ii) anti-rabbit IgG (H +
L), F(ab′)2 fragment (Alexa Fluor 555 Conjugate) (4413, Cell
Signaling technology), and (iii) goat anti-rabbit IgG (H + L) cross-adsorbed
secondary antibody, Alexa Fluor 488 (A-11008, Invitrogen). siRNAs
for PRDX2 and GAPDH were purchased from Ambion (Invitrogen, Thermo
Scientific). Lipofectamine was purchased from Invitrogen (Thermo Scientific).
An Amplex red hydrogen peroxide/peroxidase assay kit was purchased
from Invitrogen (Thermo Scientific). MTT (with 98% purity), fetal
bovine serum (FBS), trypsin (with 98% purity), PBS, glycerol (with
98.2% purity), Triton X-100 (99%), Bradford reagent, and paraformaldehyde
(with 99% purity) were obtained from Hi-Media Labs. CHEK2-proficient HCT116 human colorectal cancer cells were purchased from
NCCS, Pune. CHEK2-knockout HCT116 cells were given
as a kind gift by Dr. Bert Vogelstein from John Hopkins University.
8
Comprehensive Reagents and Cell Lines
9
PLGA Nanoparticle Synthesis and Characterization
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PLGA (Resomer® RG 503H (Mw 24,000–38,000) and Resomer® RG 752H (Mw 4000–15,000)) was obtained as a gift sample from Evonik India (Mumbai, India). Pluronics® F68 (Poloxamer 188, poly (ethylene oxide) poly (propyleneoxide) block copolymer; average molecular weight 8400), paraformaldehyde, Dulbecco’s Modified Eagle Medium (DMEM), fetal bovine serum (FBS), Trypsin Phosphate Versene Glucose (TPVG), and Dulbecco’s phosphate-buffered saline (DPBS) were purchased from Hi Media Laboratories Pvt. Ltd (Mumbai, India). 3-(4, 5-Dimethyl-2-thiazolyl)-2, 5-diphenyltetrazolium bromide was purchased form (Sigma Chemical Co., St. Louis, MO, USA). MTT reagent (3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium) was purchased from Invitrogen (Mumbai, India). 4′, 6-Diamidino-2-phenylindole (DAPI) was purchased from ThermoFisher Scientific India Pvt Ltd (Mumbai, India). Acetone, dimethyl sulfoxide (DMSO), and glacial acetic acid were purchased from SD fine chemicals (Mumbai, India). HeLa cell line was procured from National Centre for Cell Sciences (Pune, India). Deionized, double-distilled water (Milli-Q Plus system, Millipore, MA, USA) was used throughout the study. All the chemicals used were of analytical grade.
10
Mitochondrial Dynamics Analysis in Stressed Cells
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A total of 5×104 cells were seeded and grown on cover slip in a 35 mm culture dish. On reaching confluency of ~40–50%, cells were heat stressed for 30 min followed by recovery of 6 h at 37 °C. Cell incubated at 37 °C was taken as control. Following recovery cells were further incubated with 300 nM concentration of MitoTracker Orange CMTMRos (Invitrogen M7510) dissolved in DMEM (w/o FBS) for 30 min at 37 °C. Cells were washed with pre-warmed PBS and fixed using 4% paraformaldehyde in PBS (Himedia, Chennai, India) for 15 min. After fixation, cells were rinsed several times in PBS and incubated in permeabilization buffer containing 0.2% Triton X-100 for 10 min. Cells were washed with PBS thrice and counterstained with DAPI for 10 min at room temperature. Finally, coverslips were mounted onto a microscopic glass slide using ProLong Gold antifade reagent (Invitrogen P10144) and viewed under fluorescence microscope using 40× objective and a RFP/TRITC filter. Images were further analyzed using ImageJ software and ‘integrated density value’ per cell was plotted. Total of four frames were taken for analysis.
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