MA5-12023

antibody from Invitrogen Antibodies

Targeting: GFAP FLJ45472

Provider product page for MA5-12023

Western blot

Immunocytochemistry

Immunohistochemistry

Other assay

Antibody data

Antibody Data
Antigen structure
References [54]
Comments [0]
Validations
Western blot [2]
Immunocytochemistry [4]
Immunohistochemistry [5]
Other assay [13]

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Product number: MA5-12023 - Provider product page
Provider: Invitrogen Antibodies
Product name: GFAP Monoclonal Antibody (ASTRO6)
Antibody type: Monoclonal
Antigen: Other
Description: MA5-12023 targets Glial Fibrillary Acidic Protein in IF, IHC (P), and WB applications and shows reactivity with Chicken, Human, Porcine, and Rat samples. The MA5-12023 immunogen is glial Fibrillary Acidic Protein. MA5-12023 was successfully used to detect astrocytes in E18 Sparague Dawley primary cortical cells.
Reactivity: Human, Rat, Chicken/Avian, Porcine
Host: Mouse
Isotype: IgG
Antibody clone number: ASTRO6
Vial size: 500 µL
Concentration: 200 mg/mL
Storage: 4° C

GFAP protein structure - MA5-12023 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of GFAP (green) showing staining in the in the cytoplasm of SK-N-MC cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a GFAP monoclonal antibody (Product # MA5-12023) in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescent analysis of glial fibrillary acidic protein (GFAP) in E18 Sparague Dawley primary cortical neuronal cells containing astrocytes. The cells were fixed with 4% formaldehyde for 15 mins, permeabilized with 0.25% Triton X-100 in PBS for 10 mins, and blocked with 3% BSA in PBS for 30 mins at RT. Cells were stained with a GFAP mouse monoclonal antibody (Product # MA5-12023) at a dilution of 1:200 in 3% BSA in PBS for 1 hr at RT, and then incubated with Invitrogen AlexaFluor 488 Plus goat anti-mouse IgG secondary antibody (Product # A32723) at a dilution of 1:1000 for 1 hr at RT. Nuclei were stained with Hoechst 33342 (Product # H3570). The image contains overlay of GFAP (green) and nuclei (blue). Images were taken on a Zeiss LSM 710 confocal microscope at 40X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of GFAP (green) showing staining in the in the cytoplasm of SK-N-MC cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a GFAP monoclonal antibody (Product # MA5-12023) in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of GFAP (green) showing staining in the in the cytoplasm of U251 cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a GFAP monoclonal antibody (Product # MA5-12023) in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Formalin-fixed, paraffin-embedded human brain stained with GFAP antibody using peroxidase-conjugate and DAB chromogen. Note cytoplasmic staining of astrocytes.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of GFAP showing staining in the cytoplasm of paraffin-embedded human glioma (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a GFAP monoclonal antibody (Product # MA5-12023) diluted in 3% BSA-PBS at a dilution of 1:100 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of GFAP showing staining in the cytoplasm of paraffin-embedded human cerebellum tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a GFAP monoclonal antibody (Product # MA5-12023) diluted in 3% BSA-PBS at a dilution of 1:200 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of GFAP showing staining in the cytoplasm of paraffin-embedded rat brain tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a GFAP monoclonal antibody (Product # MA5-12023) diluted in 3% BSA-PBS at a dilution of 1:200 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of GFAP in human iPSC-derived astrocytes. The cells were fixed with 4% paraformaldehyde for 15 min at room temperature, and then permeabilized and blocked with 0.25% Triton X-100 and 10% donkey serum in PBS for 20 min. Samples were then incubated with a Mouse GFAP monoclonal antibody (green; Product # MA5-12023) at a dilution of 1:500 in PBS containing 0.25% Triton X-100 and 10% donkey serum at 48304C overnight, followed by incubation with Donkey anti-Mouse Alexa Fluor 488 (Product # R37114) at a dilution of 1:1,000 as well as DAPI (blue; 1:25,000) in PBS containing 0.25% Triton X-100 and 10% donkey serum at room temperature for 1 hour. Images were taken at 40X magnification. Data courtesy of Dr. Zhexing Wen at Emory University.

Supportive validation

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig 8 HSV-1 infection accelerated the astrocytes activation in neuroepithelial buds and cerebral organoids. (A) Immunofluorescence staining for the astrocyte marker (GFAP) in neuroepithelial buds at the day 18 after 3 days with HSV-1 infection. The images were taken Images were captured on a Leica TCS SP8 STED confocal microscope. (B) Relative fluorescence intensity statistics of GFAP expression were shown in different groups. (C) Detection of GFAP mRNA expression in neuroepithelial buds (n = 4). Data represent the mean +- SEM. **p < 0.01 by Student's t test. (D) The sample images of immunostaining for GFAP in the cerebral organoids at the day 45 for 3 days with HSV-1 infection (Mock, Low infection, and High infection). Scale bars: 25 mum. (E) The relative fluorescence intensity statistics of GFAP expressions were shown in different groups. Data represent the mean +- SEM from three experiments. **p < 0.01 by ANOVA.

Supportive validation

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Experimental details: FIGURE 2 Spinal cord injury (SCI) can activate the astrocytes in the olfactory bulb at 8 h post-spinal cord hemisection injury in mice. Representative images of GFAP-stained sections of the olfactory bulbs of the sham control (B) , SCI (C) , SCI + granulocyte colony-stimulating factor (G-CSF) i.p., and SCI + G-CSF oral groups. (A) Schematic illustration of the olfactory bulb (marked with dashed box) of all groups subjected to mRNA, protein, and immunofluorescence analyses. The SCI group exhibited a higher number of GFAP-positive cells [as indicated by arrow in panel (C) ] than the sham control group (B) . This indicated astrocytic activation and potential astrocyte-mediated inflammatory responses in the olfactory bulb at 8 h post-SCI. The immunofluorescence intensity of GFAP significantly decreased in the SCI + G-CSF i.p. [as indicated by arrow in panel (D) ] and SCI + G-CSF oral groups [as indicated by arrow in panel (E) ] [ (B-C) magnification 400x]. (E) Vertical bars indicate the mean +- standard error of mean) number of GFAP-stained cells in each group ( n = 3). *** P < 0.001 and ### P < 0.001.

Supportive validation

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Experimental details: FIGURE 3 Neuroinflammation in the mouse olfactory bulb at 8 h post-spinal cord injury (SCI). (A-C) The mRNA expression levels of IL-1beta (A) , IL-6 (B) , and GFAP (C) in the olfactory bulb of the sham control, SCI, SCI + granulocyte colony-stimulating factor (G-CSF) i.p., and SCI + G-CSF oral groups at 8 h post-SCI. Vertical bars indicate the mean +- standard error of the mean (SEM) ( n = 6 for each group). * P < 0.05, *** P < 0.001, ## P < 0.01, and ### P < 0.001. (D-F) The protein expression levels of IL-1beta (D) , IL-6 (E) , and GFAP (F) in the olfactory bulb of the four experimental groups at 8 h post-SCI. Representative immunoblots of IL-1beta, IL-6, GFAP, and beta-actin (internal control) are shown in the upper panel. The lower panel indicates the ratio of target protein band intensity to beta-actin protein band intensity relative to the control group (mean +- SEM). G-CSF mitigates SCI-induced neuroinflammation in the olfactory bulb as evidenced by the decreased expression of IL-1beta, IL-6, and GFAP. Vertical bars indicate mean +- SEM ( n = 6 for each group). * P < 0.05, ** P < 0.01, and ## P < 0.01.

Supportive validation

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Experimental details: 5 FIGURE Characterization of the neuro-inductive potential of meninges-derived hydrogel (MeninGEL). (a) Macroscopic images of MeninGELs after 10-day culture. (b) Inverted images of human mesenchymal stem cells (hMSCs) cultured in MeninGEL (10 mg mL -1 ) and Matrigel TM (10 mg mL -1 ) within growth medium and N2B27. (c) DAPI staining of emerged micro-tissues. (d) Immuno/histochemistry for hMSCs cultured within MeninGEL (10 mg mL -1 ) and Matrigel TM : hematoxylin and eosin, Nestin, MAP2, and GFAP staining. Gene expression study for Nestin (e), MAP2 (f), and GFAP (g) ( n = 3, * p < 0.05, ** p > 0.05)

Supportive validation

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Experimental details: FIGURE 5 Fluorescent microphotographs of GFAP immune-stained samples from control and HI piglets. GFAP reveals the length of the processes of radial-glia/neural stem cells in the SVZ (A) . HI [ (B) and graph] significantly reduced the length of GFAP + processes in the three subareas of the SVZ: roof ( ** p < 0.01), DL-SVZ ( ** p < 0.01), and L-SVZ (* p < 0.05). LV, lateral ventricle; SVZ, subventricular zone; CDT, caudate nucleus; PvWM, periventricular white matter. Original magnification 400x. Scale bar: 100 mum.

Supportive validation

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Experimental details: Fig. 6. Role of the neuronal calcium-calmodulin-dependent kinase II (CaMKII)-cyclic adenosine monophosphate response element-binding protein (CREB) cascade in spinal interleukin (IL)-33&solST2 signaling-mediated neuropathic pain. ( A and B ) Western blot analysis showing the time courses of the expressions of phosphorylated (p)CaMKII&solCaMKII and pCREB&solCREB after spared nerve injury (SNI). Representative bands are shown on the top , and a data summary is shown on the bottom . &ast P < 0.05, &ast&ast P < 0.01 compared with the naive group; n = 4 mice for each group. ( C ) Immunofluorescence image showing the pCREB-immunoreactivity (IR) in the ipsilateral and contralateral dorsal spinal cord ( top rows ) and the colocalizations of pCREB-IR with NeuN-IR ( green ), glial fibrillary acidic protein (GFAP)-IR ( green ), CD11b-IR ( green , middle rows ), and ST2 ( green , bottom rows ). The tissues were collected on the 7th day after SNI or sham operation. Scale bars = 100 &mgrm. ( D ) Western blot analysis showing the alterations in the expressions of pCaMKII&solCaMKII and pCREB&solCREB 5 h after a single intrathecal administration of the ST2 antibody (300 ng) on the 7th day after SNI. &ast P < 0.05, &ast&ast P < 0.01 compared with the naive group. &num P < 0.01 compared with the SNI &plus IgG group; n = 4 mice for each group. ( E ) The changes in the pCaMKII&solCaMKII and pCREB&solCREB levels in the spinal cords of the naive, wild-type (WT), and ST2 -&sol- mice on the 7th day a

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 7. Role of the astroglial janus kinase 2 (JAK2)-signal transducer and activator of transcription 3 (STAT3) cascade in spinal interleukin (IL)-33&solST2 signaling-mediated neuropathic pain. ( A ) Western blot analysis showing the time courses of the expressions of phosphorylated (p)JAK2&solJAK2, pSTAT3&solSTAT3, SOCS3, and glial fibrillary acidic protein (GFAP) after spared nerve injury (SNI). &ast P < 0.05, &ast&ast P < 0.01 compared with the naive group; n = 6 mice for each group. ( B ) Immunofluorescence showing the expression of pSTAT3-immunoreactivity (IR) in the superficial dorsal horn ( top rows ) and its colocalizations with NeuN-IR, GFAP-IR, CD11b-IR ( middle rows ), and ST2-IR ( bottom rows ) in the ipsilateral dorsal spinal cord. The tissues were collected on the 7th day after SNI or sham operation. Scale bars = 100 &mgrm. ( C ) Western blot analysis showing the expressions of pJAK2&solJAK2, pSTAT3&solSTAT3, SOCS3, and GFAP 5 h after a single intrathecal administration of ST2 antibody (300 ng) on the 7th day after SNI. &ast P < 0.05, &ast&ast P < 0.01 compared with the naive group; &num P < 0.01 compared with the SNI &plus IgG group; n = 6 mice for each group. ( D ) Results of the Western blot analysis of pJAK2&solJAK2, pSTAT3&solSTAT3, SOCS3, and GFAP in the spinal cords of wild-type (WT) and ST2 -&sol- mice on the 7th day after SNI. &ast P < 0.05, &ast&ast P < 0.001 compared with the naive group. &num P < 0.01 compared with the WT SNI group; n = 6 mice for ea

Supportive validation

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Experimental details: Fig 4 Histological analysis performed on paraffin-embedded slices of resected GB of a rat. A. H&E staining confirmed the presence of GB characterized by central tumor necrosis (1), tumor (2) and the peritumoral zone with infiltrating cancer cells (3) surrounded by healthy brain tissue (4). B. Immunohistochemistry for Ki67 indicated that GB is highly proliferative, except for the necrotic tumor core: tumor necrosis (1), tumor (2), peritumoral zone with infiltrating cancer cells (3) and healthy brain tissue (4). C-D. Immunohistochemistry for GFAP and Cx43 demonstrated GFAP-positive reactive astrocytes and enhanced Cx43 expression at the peritumoral zone.