Anti gfap

Manufactured by Santa Cruz Biotechnology

Sourced in United States, United Kingdom, Germany

Anti-GFAP is a laboratory reagent that binds to and detects the Glial Fibrillary Acidic Protein (GFAP), an intermediate filament protein found in astrocytes and other glial cells in the central nervous system. It is commonly used as a marker for the identification and visualization of astrocytes in various research applications.

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Lab products found in correlation

Close spelling variants of this product

Goat anti-glial fibrillary acidic protein (GFAP; (3 citations) Mouse anti-glial fibrillary acidic protein (GFAP (2 citations) Goat anti-GFAP antibody (2 citations) Mouse anti-GFAP (17 citations) Mouse monoclonal anti-GFAP (4 citations) Goat anti-GFAP (15 citations) Rabbit polyclonal anti-GFAP antibody (2 citations) Anti-GFAP antibody (7 citations) Anti-GFAP (sc-33673, (5 citations) Anti-Glial fibrillary acidic protein (GFAP; (4 citations)

57 protocols using anti gfap

Western Blot Analysis of Protein Expression

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Western blotting was performed as described in our earlier studies [26 (link),50 (link),51 (link)]. Equal amounts of proteins were electrophoresed in 10% or 12% SDS-PAGE and transferred onto a nitrocellulose membrane. The blot was probed with primary antibodies overnight at 4 °C. The following are the primary antibodies used in this study: anti-iNOS (1:1000, BD Biosciences), anti-Iba1 (1:1000, Abcam), anti-GFAP (1:1000, Santa Cruz Biotechnology, Dallas, TX, USA) and anti-β-actin (1:5000, Abcam) (Table 1). Following the overnight incubation, primary antibodies were removed, the blots were washed with phosphate buffer saline containing 0.1% Tween-20 (PBST) and corresponding infrared fluorophore-tagged secondary antibodies (1:10,000, Jackson Immuno-Research) were added at room temperature. The blots were then incubated with secondary antibodies for 1 h. Later, blots were scanned with an Odyssey infrared scanner (Li-COR, Lincoln, NE, USA). Band intensities were quantified using the ImageJ software (NIH, Bethesda, MD, USA).

Rangasamy S.B., Raha S., Dasarathy S, & Pahan K. (2021). Sodium Benzoate, a Metabolite of Cinnamon and a Food Additive, Improves Cognitive Functions in Mice after Controlled Cortical Impact Injury. International Journal of Molecular Sciences, 23(1), 192.

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Protein Expression Analysis in Mouse Brain

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The brain tissues were homogenized with lysis buffer (PROPREP; iNtRON, Sungnam, Korea; n=8 mice per group) and centrifuged at 2,500×g for 15 min at 4°C. Equal amounts of total protein (40 μg) isolated from brain tissues were resolved on 8 or 10 % sodium dodecyl sulfate polyacrylamide gels and then transferred to nitrocellulose membranes (Hybond ECL; Amersham Pharmacia Biotech, Piscataway, NJ, USA). Membranes were incubated at roomtemperature for 2 h with anti-Tyrosine hydroxylase (Cell Signaling Mol Neurobiol Technology, Inc.), anti-GFAP (1:1,000; Santa Cruz Biotechnology, Inc.) and anti-β-actin (1:2,500; Sigma-Aldrich). Blots were then incubated at room temperature for 2 h with, corresponding peroxidase-conjugated anti-mouse or anti-rabbit antibodies (1:2,000; Santa Cruz Biotechnology, Inc.). Immunoreactive proteins were detected using an enhanced chemiluminescence Western blotting detection system. The relative density of the protein bands was scanned densitometrically using My Image (SLB, Seoul, Korea) and quantified by Lab Works 4.0 (UVP, Upland, CA, USA).

Ham H.J., Yeo I.J., Jeon S.H., Lim J.H., Yoo S.S., Son D.J., Jang S.S., Lee H., Shin S.J., Han S.B., Yun J.S, & Hong J.T. (2021). Botulinum Toxin A Ameliorates Neuroinflammation in the MPTP and 6-OHDA-Induced Parkinson’s Disease Models. Biomolecules & Therapeutics, 30(1), 90-97.

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Immunohistochemical Analysis of Neurodegenerative Markers

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The immunohistochemical study was performed as previously described [57 (link),58 (link)]. Primary antibodies used were: anti-GFAP (Santa Cruz Biotechnology, 1:100 in PBS, v/v), anti-Iba1 (Santa Cruz Biotechnology, 1:100 in PBS, v/v), anti-TH (Millipore, 1:500 in PBS, v/v, Burlington, MA, USA), anti-DAT (Santa Cruz Biotechnology, 1:300 in PBS, v/v), anti-α-syn (Santa Cruz Biotechnology, 1:100 in PBS, v/v). The slices were then rinsed with PBS and treated with secondary antibody the next day. A biotin-conjugated goat anti-rabbit IgG and an avidin-biotin peroxidase complex were used to identify specific labeling (Vector). Five stained sections from each mouse were scored blindly and examined using a Leica DM6 microscope (Leica Microsystems SpA, Milan, Italy) in accordance with standard procedures [59 (link),60 (link)]. The histogram profile is related to the positive pixel intensity value obtained.

Cordaro M., D’Amico R., Fusco R., Genovese T., Peritore A.F., Gugliandolo E., Crupi R., Di Paola D., Interdonato L., Impellizzeri D., Cuzzocrea S., Di Paola R, & Siracusa R. (2022). Actaea racemosa L. Rhizome Protect against MPTP-Induced Neurotoxicity in Mice by Modulating Oxidative Stress and Neuroinflammation. Antioxidants, 12(1), 40.

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Comprehensive Biomarker Immunoassay Panel

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Anti-APP, anti-Aβ_1–42 monomer, anti-Aβ_1–42 oligomer, anti-Tau, anti-STAT3, anti-NLRP3, anti-ApoE4, anti-Nogo-A, anti-IL-13, anti-TNFα, anti-GFAP, anti-SORL1, anti- UCHL1, anti-Visfatin and anti-Clusterin were used (Santa Cruz, USA).

Park J.S., Kim S.T., Kim S.Y., Jo M.G., Choi M.J, & Kim M.O. (2019). A novel kit for early diagnosis of Alzheimer’s disease using a fluorescent nanoparticle imaging. Scientific Reports, 9, 13184.

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Molecular Markers in Neurodegeneration

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The following antibodies were used in western blot and immunofluorescence studies: p-AMPK, AMPK p-CREB (Cell Signaling), anti-Nrf-2, anti-PSD-95, anti-Syntaxin, anti-synaptosomal-associated protein 23 (SNAP-23), anti-Caspase 3, anti- PARP-1, anti-Cleaved Caspase-3, Synaptophysin, anti-Bax, anti-Bcl2, anti-TNF-α, anti-IL-1β, anti-p-NF-κB, anti-Iba-1, anti-GFAP, and anti-β-actin, which were all obtained from Santa Cruz Biotechnology (Dallas, TX, USA). Primary antibodies were diluted in 1× TBST (Tris-buffered saline plus Tween) (1:1000), and secondary conjugated anti-mouse horseradish peroxidase (HRP) and conjugated anti-rabbit HRP were diluted 1:10,000 in 1× TBST, all purchased from Promega USA. For confocal microscopic studies, the secondary fluorescent antibodies used were goat anti-mouse and goat anti-rabbit diluted in 1 × 100 phosphate-buffered saline (PBS).

Rehman S.U., Ikram M., Ullah N., Alam S.I., Park H.Y., Badshah H., Choe K, & Ok Kim M. (2019). Neurological Enhancement Effects of Melatonin against Brain Injury-Induced Oxidative Stress, Neuroinflammation, and Neurodegeneration via AMPK/CREB Signaling. Cells, 8(7), 760.

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Western Blot Analysis of Mouse Brain Proteins

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RIPA extracts from mouse brain homogenates were used for western Blot analyses as previously described^{14 (link),15 (link)}. Briefly, samples were electrophoresed on 10% Bis-Tris gels or 3–8% Tris-acetate gel (Bio-Rad, Richmond, CA), transferred onto nitrocellulose membranes (Bio-Rad) and then incubated overnight at 4 °C with the appropriate primary antibodies; anti-5LO [dilution: 1:200] (Santa Cruz, Dallas, TX), anti-HT7 [1:200] (Thermo, Waltham, MA), anti-AT8 [1:100] (Thermo), anti-AT270 [1:200] (Thermo), anti-PHF13 [1:100 (Thermo)], anti-SYP [1:300] (Santa Cruz), anti-PSD95 [1:200] (Thermo), anti-GSK3α/β [1:100] (Cell Signaling, Danvers, MA), anti-pGSK3α/β [1:100] (Cell Signaling), anti-SAPK/JNK [1:100] (Cell Signaling), anti-pSAPKJNK [1:100] (Cell Signaling), anti-cdk5 1[:200] (Santa Cruz), anti-p35/p25 [1:100] (Santa Cruz), anti-PP2A [1:200] (Santa Cruz), anti-GFAP (Santa Cruz), anti-Iba1[1:100] (Thermo) and anti-Beta actin [1:500] (Santa Cruz). After three washings with T-TBS (pH 7.4), membranes were incubated with IRDye 800CW-labeled secondary antibodies (LI-COR Bioscience, Lincoln, NE) at room temperature for 1 h. Signals were developed with Odyssey Infrared Imaging Systems (LI-COR Bioscience, Lincoln, NE). β-Actin was always used as an internal loading control.

Vagnozzi A.N., Giannopoulos P.F, & Praticò D. (2017). The direct role of 5-lipoxygenase on tau pathology, synaptic integrity and cognition in a mouse model of tauopathy. Translational Psychiatry, 7, 1288.

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Immunofluorescence Staining of Brain Tissues

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Briefly, after being anesthetized with isoflurane and intracardially perfused with PBS and 10% formalin, brains were removed, fixed in 10% formalin overnight, immersed in 30% sucrose for 3 d, and then frozen. Fixed frozen brain tissues were sectioned at 10 μm by using cryostat (Leica CM3050S-3-1-1, IL). Double immunofluorescence staining was performed as previously reported (He et al. 2015 (link)). Sections were permeabilized with 0.3% Triton X-100 for 30 min. After blocking with 5% donkey serum for 1h, the sections were incubated with the following primary antibodies: anti-MSP (1:1000, Abcam, MA, RRID: AB_10976147), anti-RON (1:1000, Abcam, MA, RRID: AB_10972503), anti-GFAP (1:1000, Santa Cruz Biotechnology, TX, RRID: AB_627673), anti-vWF (1:500, Santa Cruz Biotechnology, TX, RRID: AB_10842026) and anti-NeuN (1:500, Abcam, MA, RRID: AB_2532109), at 4℃ overnight, followed by appropriate fluorescence-conjugated secondary antibodies (Jackson ImmunoResearch Laboratories, PA, RRID: AB_2337972, AB_ 2338059, AB_2340432 or AB_2338871) at room temperature for 2h. The co-localizations were evaluated by a fluorescent microscope (Olympus OX51, Tokyo, Japan).

Lu T., Wang Z., Prativa S., Xu Y., wang T., Zhang Y., Yu L., Xu N., Tang J., You W., Chen G, & Zhang J.H. (2018). Macrophage Stimulating Protein (MSP) Preserves Blood Brain Barrier Integrity After Intracerebral Hemorrhage Through RON Dependent GAB1/Src/β-catenin Pathway Activation in a Mouse Model. Journal of neurochemistry, 148(1), 114-126.

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Spinal Cord Protein Expression

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All spinal cord tissues (T10 spinous process) obtained at 7, 14, 21, and 28 days were homogenized in lysis buffer (Kangwei Biotechnology, Beijing, China). Equal amounts of proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Resolved proteins were transferred to a nitrocellulose membrane (Whatman, Dassel, Germany), which was incubated with rabbit anti-GFAP (1:1000), goat anti-P2X4 (1:1000), and anti-β-actin (1:2000; Santa Cruz Biotechnology, Dallas, TX, USA) antibodies overnight at 4°C. Subsequently, membranes were rewarmed, washed, and horseradish peroxidase was added to label the secondary antibody. The β-actin antibody was used as a loading control for all experiments. Immunoreactive bands were visualized with an enhanced chemiluminescence reagent (Beyotime, Beijing, China). Grayscale values of bands were quantified with ImageJ software (NIH). Relative protein expression levels were calculated according to the ratio between target grayscale values and loading control grayscale values.

Du X.J., Chen Y.X., Zheng Z.C., Wang N., Wang X.Y, & Kong F.E. (2019). Neural stem cell transplantation inhibits glial cell proliferation and P2X receptor-mediated neuropathic pain in spinal cord injury rats. Neural Regeneration Research, 14(5), 876-885.

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Alzheimer's Biomarker Expression Analysis

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Alpha-linoleic acid (cis-9,cis-12-octadecadienoic acid, Lot#30H8479) and Aβ_1–42 peptides were purchased from Sigma-Aldrich Chemical Co., St. Louis, MO, USA. The antibodies used in the study included anti-Aβ (sc-28365), anti-p-Tau (sc-390476), anti-BACE-1 (sc-33711), p-Tau (sc-)anti-PSD-95 (sc-71933), anti-synaptosomal-associated protein 23 (SNAP-23) (sc-374215), anti-p-JNK (sc-6254), anti-Caspase 3 (sc-7272), anti-Bax (sc-7480), anti-Bcl-2 (sc-7382), anti-PARP-1 (sc-7008), anti-TNF-α (sc-52746), anti-p-NF-κB p65 (Ser536) (sc-136548), anti-TLR4 (sc-293072) anti-Iba-1 (sc-32725), anti-GFAP (sc-33673), and anti-β-actin (sc-47778) from Santa Cruz Biotechnology (Dallas, TX, USA). The primary antibodies were diluted in TBST (1:1000) (Santa Cruz Biotechnology) and the secondary anti-mouse HRP conjugated (Promega Ref# W402) and antirabbit HRP conjugated (Promega Ref# W401) antibodies that were diluted to 1:10,000 in 1 × TBST were obtained from Promega, USA. For confocal microscopic studies, the secondary fluorescent antibodies used were goat anti-mouse (Ref# A11029) and goat anti-rabbit (Ref# 2732), which were diluted in 1× phosphate-buffered saline (PBS).

Ali W., Ikram M., Park H.Y., Jo M.G., Ullah R., Ahmad S., Abid N.B, & Kim M.O. (2020). Oral Administration of Alpha Linoleic Acid Rescues Aβ-Induced Glia-Mediated Neuroinflammation and Cognitive Dysfunction in C57BL/6N Mice. Cells, 9(3), 667.

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Immunocytochemical Staining of Cells

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Cells were fixed with paraformaldehyde
(4%) for 1 h at room temperature
and permeabilized with Triton X-100 (0.2%, 10 min), followed by blocking
with bovine serum albumin (2%). Fixed cells were incubated at 4 °C
overnight with the primary antibodies including anti-MAP-2 (1/500,
Sigma, St. Louis, MO), anti-GFAP (1/1000, Santa Cruz, CA), anti-EPO
and EPOR (Santa Cruz, CA). Cells were incubated with a fluorescence-conjugated
secondary antibody at room temperature (2 h, Alexa Fluor 488 donkey
anti-mouse IgG, 1:1000; donkey anti-rabbit IgG, Molecular Probes,
Leiden, Netherlands). Cells with a secondary antibody only were used
as a negative control.

Samson F.P., He W., Sripathi S.R., Patrick A.T., Madu J., Chung H., Frost M.C., Jee D., Gutsaeva D.R, & Jahng W.J. (2020). Dual Switch Mechanism of Erythropoietin as an Antiapoptotic and Pro-Angiogenic Determinant in the Retina. ACS Omega, 5(33), 21113-21126.

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