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Anti gfap

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Anti-GFAP is a primary antibody that specifically recognizes the Glial Fibrillary Acidic Protein (GFAP), which is a type III intermediate filament protein expressed in astrocytes and other glial cells in the central nervous system. This antibody can be used for the detection and localization of GFAP in various research applications.

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Lab products found in correlation

Anti gapdh, by GeneTex (2 mentions) Bx51trf, by Olympus (2 mentions) Anti olig2, by GeneTex (2 mentions) Anti iba1, by Fujifilm (2 mentions) Alexa fluor 568, by Thermo Fisher Scientific (2 mentions) Anti cd4, by Abcam (1 mentions) Prolong diamond antifade mountant, by Thermo Fisher Scientific (1 mentions) Axio vert a1 microscope, by Zeiss (1 mentions) Anti coliv, by Bio-Rad (1 mentions) Actingreen 488 readyprobes reagent, by Thermo Fisher Scientific (1 mentions) Anti iba1, by Abcam (1 mentions) Hyclone, by Thermo Fisher Scientific (1 mentions) Anti α sma, by Merck Group (1 mentions) Anti ccnd1, by Cell Signaling Technology (1 mentions) Anti phospho akt ser473, by Cell Signaling Technology (1 mentions) Anti akt, by Cell Signaling Technology (1 mentions) Anti cdk4, by Cell Signaling Technology (1 mentions) Anti phospho erk1 2 thr202 tyr204, by Cell Signaling Technology (1 mentions) Anti top2a, by Cell Signaling Technology (1 mentions) Anti cdk2, by GeneTex (1 mentions)

Spelling variants of this product

Rabbit anti-GFAP (4 citations) Chicken anti-GFAP (3 citations)

6 protocols using anti gfap

1

Immunostaining Workflow for Tissue Analysis

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Immunostaining was performed on paraffin sections. After paraffin removal, sections were rehydrated and blocked for 30 minutes at room temperature in 1× PBS solution containing 10% fetal calf serum (HyClone; Thermo Fisher Scientific) and 0.3% Triton X-100. Sections were then incubated overnight at 4°C with rabbit primary antibodies diluted in blocking solution, washed in 1× PBS supplemented with 0.1% Tween-20, incubated for 2 hours at room temperature with Donkey anti-Rabbit secondary antibody, Alexa Fluor 568 (#A10042, 1:500; Thermo Fisher Scientific), washed and mounted in ProLong Diamond Antifade Mountant (#P36961; Thermo Fisher Scientific). Primary antibodies and probes used in this study included anti-GFAP (#GTX108711, 1:200; GeneTex, Inc., Irvine, CA, USA); anti-Iba1 (#AB178846, 1:500; Abcam, Cambridge, UK); anti-α-SMA (#C6198, 1:500; Sigma-Aldrich, St. Louis, MO, USA); anti-Col-IV (#2150-1470, 1:200; Bio-Rad Laboratories); anti-CD4 (#183685, 1:200; Abcam); and ActinGreen 488 ReadyProbes Reagent (#R37110; Thermo Fisher Scientific).
Immunostaining was observed using an AxioVert.A1 microscope (Carl Zeiss Microscopy) equipped with a QImaging Retiga R3 camera (QImaging, Surrey, BC, Canada) and an X-Cite series 120 fluorescence lamp (Excelitas Technologies, Waltham, MA, USA). Images were documented using the Fiji and µManager software packages.
Lhussiez V., Dubus E., Cesar Q., Acar N., Nandrot E.F., Simonutti M., Audo I., Lizé E., Nguyen S., Geissler A., Bouchot A., Ansar M., Picaud S., Thauvin-Robinet C., Olivier-Faivre L., Duplomb L, & Da Costa R. (2020). Cohen Syndrome-Associated Cataract Is Explained by VPS13B Functions in Lens Homeostasis and Is Modified by Additional Genetic Factors. Investigative Ophthalmology & Visual Science, 61(11), 18.
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2

Immunoblotting and Immunohistochemical Analysis

Cited 1 time
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Immunoblotting was performed using anti-phospho-RB (Cell Signaling, Danvers, MA, USA), anti-RB (GeneTex, Irvine, CA, USA), anti-phospho-Akt(Ser473) (Cell Signaling), anti-Akt (Cell Signaling), anti-phospho-Erk1/2(Thr202/Tyr204) (Cell Signaling), anti-Erk1/2 (Cell Signaling), anti-TOP2A (Cell Signaling), anti-CCND1 (Cell Signaling), anti-γH2AX (Cell Signaling), anti-CDK2 (GeneTex), anti-RAD51 (GeneTex), anti-CDK4 (Cell Signaling), and anti-GAPDH (GeneTex) antibodies followed by horseradish peroxidase-conjugated secondary antibodies and visualization using an enhanced chemiluminescence detection system. Immunohistochemical (IHC) sample preparation and staining with anti-GFAP (Genetex), anti-phospho-RB (Cell Signaling), anti-Ki-67 (Cell Signaling), and anti-γH2AX (Cell Signaling) antibodies were carried out as previously described [48 (link)]. Uncropped Western blot images are provided in Figure S5.
Liang M.L., Chen C.H., Liu Y.R., Huang M.H., Lin Y.C., Wong T.T., Lin S.E., Chu S.S., Ding Y.H, & Hsieh T.H. (2020). Abemaciclib, A Selective CDK4/6 Inhibitor, Restricts the Growth of Pediatric Ependymomas. Cancers, 12(12), 3597.
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3

Immunohistochemical Analysis of Spinal Cord Injury

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After the pigs were sacrificed, spinal cord sections (1 cm) from the central DSCI lesion region were collected and embedded in paraffin. Spinal cord sections (5 μm thick) from each specimen were deparaffinized with xylene and incubated in graded concentrations of ethanol. They were then washed with phosphate-buffered saline (PBS) for 3 × 5 min. The sections were incubated for blocking with a blocking solution (0.1% Triton X-100 in PBS and 10% normal goat serum) at room temperature for 2 h. The sections were incubated overnight with primary antibodies at 4°C. The primary antibodies used were anti-GFAP (1: 2000, GeneTex), anti-Iba-1 (1:200, Affinity), and anti-NeuN (1:100, Abcam). After rinsing with PBS for 3 × 5 min, the sections were incubated with secondary antibodies for 2 h at room temperature. The secondary antibody used in this study was goat anti-rabbit antibody (Alexa Fluor®594) (1:200, Abcam). Following three rinses with PBS, a drop of antifade mounting medium containing DAPI (Solarbio Biotechnology, China) was placed on each slide. Finally, a coverslip was placed on top of each sample. Immunofluorescence imaging was carried out using an Olympus fluorescence microscope.
Han B., Liang W., Hai Y., Liu Y., Chen Y., Ding H., Yang J, & Yin P. (2022). Elucidating the Potential Mechanisms Underlying Distraction Spinal Cord Injury-Associated Neuroinflammation and Apoptosis. Frontiers in Cell and Developmental Biology, 10, 839313.
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4

Protein Quantification and Western Blot Analysis

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To assess protein level in brain or cells, lysates were prepared in SDS lysis buffer containing protease inhibitors and phosphatase inhibitors. The amount of proteins in lysates were normalized by BCA protein assay kit (Millipore). Proteins were resolved by SDS–polyacrylamide gel electrophoresis and then transferred to nitrocellulose membranes (PerkinElmer). Transferred blots were incubated with primary antibodies, anti-PGAM5 (1:500, Santa Cruz, SC-515880), anti-GAPDH (1:5000, Genetex, GTX100118), anti-TUJ1 (1:5000, BioLegend, #801202), anti-GFAP (1:5000, Genetex, GTX108711), anti-PGC1α (1:500, Genetex, GTX37356), anti-NRF1 (1:1000, Genetex, GTX103179), anti-TFAM (1:500, Genetex, GTX59889), or anti-TIM23 (1:500, Santa Cruz, SC-514463) overnight. Then the blots were incubated with secondary antibodies, IRDye 800CW goat anti-rabbit IgG secondary antibody (1:1000, LI-COR, # 926-32211) or goat anti-mouse IgG secondary antibody Alexa Fluor™ 700 (1:1000, Invitrogen, # A-21036), for 1 h. Membranes were imaged by ChemiDoc™ MP Imaging System (Bio-Rad) and the signal intensity of bands was quantified by Image Lab software (Bio-Rad, version 6.1.0) (Additional file 2).
Liang M.Z., Lu T.H, & Chen L. (2023). Timely expression of PGAM5 and its cleavage control mitochondrial homeostasis during neurite re-growth after traumatic brain injury. Cell & Bioscience, 13, 96.
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5

Immunohistochemical Analysis of Brain Tissue

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Firstly, brains were xated with 4% PFA, and they were embedded in para n after dehydration. Coronal sections of brain tissues were cut by a microtome (Microm, Walldorf, Germany) for antigen retrieval, and immersed into citrate buffer before being permeabilized by 20% tween for 30 min. For nonspeci c markers blocking, we used 0.1% BSA in 0.1% triton for 1 h. Then, sections were incubated at 4°C with primary antibodies: anti-Iba-1 (1:250; Wako chemicals, Virginia), anti-GFAP (1:100; Gene Tex, CA) and anti-Olig2 (1:100; Gene Tex, CA) overnight. After the addition of secondary antibodies (Vector Labs, CA), slides were washed with PBS twice; the nuclei were stained with DAPI (4', 6-diamidino-2-phenylindole) for 1 min. Finally, slides were assessed by a uorescent microscope (Olympus BX51TRF, Tokyo, Japan) equipped with a digital camera (Olympus) for capturing images.
Tahmasebi F., Barati S, & Kashani I.R. (2021). Effect of CSF1R inhibitor on glial cells population and remyelination in the cuprizone model. Neuropeptides, 89.
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6

Immunohistochemical Analysis of Brain Tissue

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Firstly, brains were xated with 4% PFA, and they were embedded in para n after dehydration. Coronal sections of brain tissues were cut by a microtome (Microm, Walldorf, Germany) for antigen retrieval, and immersed into citrate buffer before being permeabilized by 20% tween for 30 min. For nonspeci c markers blocking, we used 0.1% BSA in 0.1% triton for 1 h. Then, sections were incubated at 4°C with primary antibodies: anti-Iba-1 (1:250; Wako chemicals, Virginia), anti-GFAP (1:100; Gene Tex, CA) and anti-Olig2 (1:100; Gene Tex, CA) overnight. After the addition of secondary antibodies (Vector Labs, CA), slides were washed with PBS twice; the nuclei were stained with DAPI (4', 6-diamidino-2-phenylindole) for 1 min. Finally, slides were assessed by a uorescent microscope (Olympus BX51TRF, Tokyo, Japan) equipped with a digital camera (Olympus) for capturing images.
Tahmasebi F., Barati S., & Kashani I.R. (2021). Effect of CSF1R Inhibitor on Glial Cells Population and Remyelination in The Cuprizone Model.
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