The CEBPD, p67phox, and nitrotyrosine antibodies and TETA were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The GFAP antibody was purchased from Invitrogen (Carlsbad, CA, USA). The p47phox antibody was purchased from MDBio Inc. (Taipei, Taiwan). The SOD1 antibody was purchased from Abcam plc. (Cambridge, MA). The Dulbecco's modified Eagle's medium (DMEM), TRIzol RNA extraction reagent, and SuperScript™ III were purchased from Invitrogen (Carlsbad, CA, USA). Fetal bovine serum (FBS) was purchased from HyClone Laboratories (Logan, UT, USA). All oligonucleotides were synthesized by MDBio Inc. (Taipei, Taiwan).
Gfap antibody
Manufactured by Thermo Fisher Scientific
Sourced in United States
The GFAP antibody is a laboratory reagent used to detect the presence and levels of the glial fibrillary acidic protein (GFAP) in biological samples. GFAP is an intermediate filament protein expressed in astrocytes and other glial cells. The GFAP antibody can be used in various analytical techniques, such as Western blotting, immunohistochemistry, and flow cytometry, to study the expression and distribution of GFAP in different tissues and cell types.
Automatically generated - may contain errors
Lab products found in correlation
Sodium bicarbonate, by Thermo Fisher Scientific (3 mentions)
Trypsin edta, by Thermo Fisher Scientific (3 mentions)
Dnase, by Thermo Fisher Scientific (3 mentions)
Trizol rna extraction reagent, by Thermo Fisher Scientific (2 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (2 mentions)
Superscript 3, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Cytiva (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
Fluo 4 am, by Thermo Fisher Scientific (2 mentions)
P67phox, by Santa Cruz Biotechnology (1 mentions)
Cebpd, by Santa Cruz Biotechnology (1 mentions)
Sod1 antibody, by Abcam (1 mentions)
Znf179 antibody, by GeneTex (1 mentions)
Caspase 3 antibody, by Cell Signaling Technology (1 mentions)
Mouse on mouse polymer ihc kit, by Abcam (1 mentions)
Te300 microscope, by Nikon (1 mentions)
Fluorophore labeled secondary antibodies, by Jackson ImmunoResearch (1 mentions)
Image pro software, by Media Cybernetics (1 mentions)
Tissue tek oct, by Sakura Finetek (1 mentions)
9 protocols using gfap antibody
1
Oxidative Stress Signaling Pathway
2
Immunoblotting and Immunofluorescence Assays
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The CEBPD and green fluorescent protein (GFP) antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The GFAP antibody was purchased from Invitrogen (Carlsbad, CA, USA). The caspase 3 antibody was purchased from Cell Signaling Technology (Danvers, MA, USA). The ZNF179 antibody used for immunofluorescence was purchased from GeneTex (Irvine, CA, USA), and the ZNF179 antibody used in the Western blot and immunoprecipitation assays was obtained from Dr. Yi-Chao Lee. The TRIzol RNA extraction reagent, Dulbecco’s modified Eagle’s medium (DMEM), and SuperScript™ III were purchased from Invitrogen (Carlsbad, CA, USA). All oligonucleotides were synthesized by MDBio Inc. (Taipei, Taiwan). Fetal bovine serum (FBS) was purchased from HyClone Laboratories (Logan, UT, USA).
3
Immunohistochemical Analysis of CD59a and GFAP in Mouse Eyes
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Mice were humanely euthanized, and eyes enucleated and immersion-fixed in 4% paraformaldehyde (PFA) for 10 min. Eyeballs were then rinsed in PBS, and eye cups were generated by removing the anterior segment. The eye cups were infiltrated in 30% sucrose overnight and embedded in Tissue-Tek OCT (Sakura Finetek, Torrance, CA, USA). Immunostaining for CD59a was performed on 10-μm-thick cryostat sections. The primary antibodies were mouse anti-mouse CD59a (HM1116, Hycult, PA, USA) at 1:500 dilution and a mouse-on-mouse polymer IHC kit (ab127055, Abcam, MA, USA) was used to optimize signal to background staining ratio. We used an HRP-conjugated secondary antibody followed by diaminobenzidine to yield a brown precipitate. Rat anti-mouse glial fibrillary acid protein (GFAP) antibody (13–0300, Invitrogen, Camarillo, CA, USA) was used at a 1:500 dilution. GFAP antibody was detected using fluorophore-labeled secondary antibodies (Jackson ImmunoResearch Laboratories, Inc., PA, USA). Control sections were treated identically but without primary antibody. The sections were analyzed by fluorescence microscopy with identical exposure parameters (model TE300 microscope, Nikon, Tokyo, Japan) with ImagePro software (Media Cybernetics, Silver Spring, MD, USA).
4
Immunohistochemical Analysis of Tissue Sections
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Formaldehyde-fixed tissue was embedded in paraffin wax for further immunohistochemical examination. Sections were deparaffinised in xylene. Antigen retrieval process was performed twice in citrate buffer solution (pH: 6.0), first for 7 min, and second for 5 min, boiled in a microwave oven at 700 W. They were allowed to cool to room temperature for 30 min and washed twice in distilled water for 5 min. Endogenous peroxidase activity was blocked in 0.1% hydrogen peroxide for 20 min. Ultra V block (Cat. No:85-9043, Invitrogen, Carlsbad, CA, USA) was applied for 10 min prior to the overnight application of primary antibodies VEGF antibody (Cat. No: RB-222-P0) (1:100 dilution), GFAP antibody (1:100 dilution) (Cat. No: PA3-067, Invitrogen, Carlsbad, CA, USA). Secondary antibody (Cat. No: 85-9043, Invitrogen, Carlsbad, CA, USA) was applied for 20 min. Slides were then exposed to streptavidin-peroxidase for 20 min. Chromogen diaminobenzidine (DAB) (Invitrogen, Cat. No: 34002 Carlsbad, CA, USA) was used. Control slides were prepared as mentioned above, but omitting the primary antibodies. After counterstaining with haematoxylin, and washing in tap water for 8 min and in distilled water for 10 min, the slides were mounted with Entellan.
5
Astroglial Alteration Evaluation in Striatum
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Deparaffinized 5-μm-thick tissue sections were cut and prepared for evaluation of astroglial alteration. Striatal sections were treated with 3% hydrogen peroxide for 20 min, washed with PBS, then incubated with mouse monoclonal glial fibrillary acidic protein (GFAP) antibody (Thermo Fisher Scientific Inc., United States) for 30 min. The sections were washed with PBS followed by incubation for 20 min with secondary antibody (Dako, Carpenteria, CA, United States), and then with horseradish peroxidase using the HRP Envision kit (Dako, Carpenteria, CA, United States). The reaction was visualized with 3,3′-diaminobenzidine tetrahydrochloride (DAB Substrate Kit, Vector Laboratories Inc., Burlingame, CA, United States) for 10 min following another wash with PBS. Finally, the sections were counterstained with hematoxylin, dehydrated, and cleared in xylene then cover-slipped for microscopic analysis. Six randomly selected fields from striatum region were analyzed for determination of GFAP immunoreactive percentage areas in individual sections using full HD microscopic camera operated by Leica application module for histological analysis (Sayed et al., 2020 (link)).
6
Investigating the NF-κB Pathway
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For investigation of the NF-κB pathway, IκBα, p65, and phospho-p65 antibodies were obtained from Cell Signaling Technology (Danvers, MA, USA). Vimentin, nestin, and GAPDH antibodies were obtained from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). The Human Quantikine TNF-α ELISA kit was obtained from R&D Systems (Minneapolis, MN, USA). The GFAP antibody and RIPA Cell Lysis buffer were purchased from ThermoFisher Scientific (Fair Lawn, NJ, USA). GSK1016790A (GSK101) was purchased from Sigma-Aldrich (St. Louis, MO, USA).
7
Zika Virus Infection Cell Culture
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DMEM, MEM, Neurobasal media, N2 supplement, penicillin/streptomycin, trypsin-EDTA, DNAse, HEPES solution, sodium bicarbonate, non-essential amino acid solution, GFAP antibody, and Prolong Anti-fade with DAPI were all purchased from Thermo Fisher Scientific (Waltham, MA, USA). Fetal bovine serum was purchased from Atlanta biologicals (Lawrenceville, GA, USA). Zika NS1 rabbit polyclonal antibody (GTX133306) was purchased from Genetex (San Antonio, TX, USA). Zika Virus strain PA259459 was obtained from the World Reference Center for Emerging Viruses and Arboviruses through the University of Texas Medical Branch (UTMB) at Galveston (Galveston, TX, USA). All other materials were purchased from Sigma-Aldrich (St. Louis, MO, USA).
8
Quantifying Glial Scar Formation in SCI Mice
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To quantify glial scar area in the mNPC, pLXSN-mNPC, and hADC-mNPC mice, tissue sections (20 mm) were obtained 2 weeks after SCI and sequentially immunoreacted with GFAP antibody (1:500, Thermo, Waltham, MA, USA) at 4 °C overnight. Subsequently, tissue sections were incubated with the appropriate biotinylated secondary antibodies. Immunostaining was performed using an ABC kit (Vector, Burlingame, CA, USA), followed by reaction with 3,39-diaminobenzidine tetra hydrochloride (DAB, Sigma, St. Louis, MO, USA). Negative controls lacked the primary antibody. Glial scar area was obtained from measuring GFAP-positive regions around the lesion site (with central cavity and the number of reactive astrocytes) in Image J (National Institutes of Health, Bethesda, MD, USA).
9
Iron Homeostasis in Glial Cells
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The culture medium including DMEM and fetal bovine serum were purchased from Gibco Life Technology Invitrogen (Grand Island, NY, USA). Oligo-fectamine, MEMI, fluo-4 AM, sodium-binding benzofuran isophthalate (SBFI) AM, G-agarose bead, TFR antibody, β-actin antibody, GFAP antibody, DMT1 antibody and DMT1 siRNA duplex chains were from Thermo Fisher Scientific (Waltham, MA USA); siRNA duplex chains of TFR, NCX1-3, Cav-3 and Dab2, NCX2 antibody, Na+/K+-ATPase alpha1/2 antibody and the secondary antibodies were bought from Santa Cruz Biotechnology (Santa Cruz, CA, USA). NCX1 antibody, NCX3 antibody and native mouse apo-transferrin (apo-TF; i.e., iron-free) were from Abcam (Cambridge, MA, USA). Donkey serum, xestospongin C (Xe-C), nifedipine, ferrous sulfate heptahydrate (FeSO4), sulforhodamine 101 (SR101) and ferric ammonium citrate were purchase from Sigma-Aldrich (St. Louis, MO, USA). Ryanodine and KB-R7943 were purchased from Calbiochem (La Jolla, CA, USA). Secondary antibody staining with donkey anti-mouse or anti-rabbit Cy-2/3 were from Jackson Immuno-Research (West Grove, PA, USA). Primary antibody of histone H3 was purchased from EarthOx (Millbrae, CA, USA).
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