PA5-16291

antibody from Invitrogen Antibodies

Targeting: GFAP FLJ45472

Provider product page for PA5-16291

Western blot

Immunocytochemistry

Immunohistochemistry

Other assay

Antibody data

Antibody Data
Antigen structure
References [28]
Comments [0]
Validations
Western blot [1]
Immunocytochemistry [1]
Immunohistochemistry [1]
Other assay [19]

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Product number: PA5-16291 - Provider product page
Provider: Invitrogen Antibodies
Product name: GFAP Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Other
Description: PA5-16291 targets Glial Fibrillary Acidic Protein in IHC (P) applications and shows reactivity with Bovine, Chicken, Guinea Pig, Hamster, Human, mouse, Non-human primate, Ovine, and Rat samples.

GFAP protein structure - PA5-16291 shown in red.

Supportive validation

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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 2 ( a ) Ischemia/Reperfusion (IR) induced strong activation of Muller cells, the main glia in the retina, assessed by GFAP radial staining after 3-days and persisted for 14-days after ischemia. ( b ) Immunostaining studies using anti-TXNIP (green), anti-glutamine synthetase (GS, red) showed prominent colocalization (yellow) of TXNIP within Muller cells in response to IR when compared to shams.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 5 Assessing reactive gliosis twelve months after radiation exposure. ( A ) Comparing immunohistochemical staining of nestin in cerebral cortex after radiation. ( B ) Quantification of nestin staining in cerebral cortex presented as mean +- SEM. ( C ) Comparing immunohistochemical staining of GFAP in cerebral cortex after radiation. ( D ) Quantification of GFAP staining in cerebral cortex presented as mean +- SEM.

Supportive validation

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Experimental details: Figure 2 The prostate cell lines LN-NE and LN-FLU display phenotypic features of stem cells. ( A ) Levels of the epithelial marker E-cadherin and the proangiogenic factor VEGF in androgen-depleted cell lines determined by Western blot and limited to actin as a load control. A representative image of three experiments is shown. Densitometric analysis (n = 3) is shown on the right. ( B ) LNCaP, LN-NE, and LN-FLU cells were incubated in differentiation media for 15 days, and levels of the glial marker GFAP or the osteoblast marker Osx were determined by Western blot. Levels of expression of the osteoblast markers OSX and IBSP determined by qPCR. The data show the relative mRNA expression to GAPDH, which was used as a housekeeping gene. Data represent the mean +- SD of two independent experiments. * p < 0.05 significant difference between LNCaP and LN-NE cells or LNCaP and LN-FLU cells by two-way ANOVA and Sidak's multiple comparisons test.

Supportive validation

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Experimental details: Figure 4 Mfn2 overexpression inhibited MPTP-induced astrocytosis and microgliosis in the SN. Fixed brains of MPTP- or saline-treated mice were immunostained for iba1 and GFAP. Representative pictures of microglia ( A ) and astrocytes ( B ) are shown. Quantification of the relative number of microglia ( C ) and astrocytes ( D ) in the SN (dashed line) is shown ( n = 4; * p < 0.05 compared to the control. n.s., not significant).

Supportive validation

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Experimental details: Figure 7 Mfn2 overexpression inhibited MPTP-induced inflammation in the striatum. Representative pictures of iba1 ( A ) staining in the striatum and quantification of total ( C ) or relative activated ( D ) microglia in the striatum are shown (Red arrows indicate activated microglia). Representative pictures of GFAP staining in the striatum ( B ) and quantification of astrocytes ( E ) in the striatum are shown ( n = 4; * p < 0.05, ** p < 0.01, and *** p < 0.001 compared to the control).

Supportive validation

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Experimental details: FIGURE 8 Perivascular GFAP processes are related to altered CLN-5 expression and structural deterioration of the brain vessels in human AD. (A) Representative images of triple immunolabeling of GFAP, CLN-5 and the endothelial marker UEA I showing increased astrocytic reactivity near the vessels, and augmented CLN-5 expression in SAD and FAD. (B) 2X magnifications of the insets in panel (A) highlighting the augment of both perivascular GFAP processes and vascular CLN-5 in AD. (C) Segmented images of GFAP processes present between 0 and 2 mum away from the vessel surface showing the proximity of CLN-5 with vascular areas covered by GFAP. White arrowheads indicate regions where high GFAP vascular coverage coincides with high CLN-5 staining, while empty arrowheads point to areas where both GFAP and CLN-5 are scarce. The insets are 2X magnifications of double positive GFAP-CLN-5 areas. (D) Heat map representation of superposed UEA I signals indicating structural alteration as intensity values go down. Note that the boxes in panel (D) that are enclosing vascular regions with low intensity values, correspond to the images shown in panel (C) which display increased GFAP and CLN-5. (E) Quantification of the perivascular volume of GFAP, vascular CLN-5, and perivascular double positive GFAP-CLN-5. * Comparison with CNT, levels of significance were set to * p < 0.05 and ** p < 0.01. # Comparison SAD with FAD, levels of significance were set to # p < 0.05. (F) Quantification of the Z proj

Supportive validation

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Experimental details: Figure 3 Effect of naringenin (Nar) on senescence of mouse neural stem cells (mNSCs). ( A ) p16 ink4a mRNA expression was measured by qRT-PCR in 23M-NSCs with and without treatment of 6.8 mug/mL Nar for 48 h. ( B ) Cell cycle phase distributions of 23M-NSCs treated with Nar for 48 h and control cells. ( C ) The relative telomere length of 23M-NSCs increased significantly with Nar treatment. ( D ) Immunofluorescence Ki67 staining of mNSCs treated with Nar for 48 h, with DAPI nuclear labeling. ( E ) Quantification of ( D ). ( F ) Representative fields of MAP2 and GFAP immunofluorescence staining of cultured mNSCs after control and Nar treatment. ( G ) Quantification of ( F ). Data are presented as the mean +- SD of three independent experiments. * P < 0.05, ** P < 0.01, and *** P < 0.0001 compared with untreated cells.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 2 ( a ) Ischemia/Reperfusion (IR) induced strong activation of Muller cells, the main glia in the retina, assessed by GFAP radial staining after 3-days and persisted for 14-days after ischemia. ( b ) Immunostaining studies using anti-TXNIP (green), anti-glutamine synthetase (GS, red) showed prominent colocalization (yellow) of TXNIP within Muller cells in response to IR when compared to shams.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 5 ( a ) Ischemia/Reperfusion (IR) induced strong activation of Muller cells, the main glia in the retina, assessed by GFAP radial staining after 3-days in WT-IR but not in TKO-IR when compared to shams. ( b ) Ischemia/Reperfusion (IR) sustained retinal cell death indicated by TUNEL-positive cells (arrows) after 3-days in WT-IR, but not in TKO-IR when compared to shams.

Supportive validation

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Experimental details: FIGURE 6 Increased MPTP vulnerability of VPS35 D620N KI mice. (a) A schematic diagram depicts the experimental design. (b-c) Exacerbated motor function deficits in VPS35 D620N / D620N mice as measured by the latency to fall in rotarod test (b) and duration to cross the narrow beam (9 mm) in beam walking test (c) ( n = 12-14/group). (d-g) Concentrations of DA (d), DOPAC (e), HVA (f), and DA turnover rate (DOPAC+HVA/DA) (g) in the STR of VPS35 D620N / D620N and WT controls injected with MPTP/Saline as indicated ( n = 10-12/group). (h) Representative images of TH, GFAP, and IBA1 immunostaining in SNpc and STR from VPS35 D620N / D620N and WT mice injected with MPTP/Saline as indicated. Scale bar, 100 mum. Quantification of TH-positive neurons in SNpc (i), OD of TH-positive fibers in STR (j), GFAP-positive cells (k) and IBA1-positive cells (l,m) in SNpc ( n = 9/group). Representative Western blots (n) and quantification (o) of TH in VM extracts from VPS35 D620N / D620N mice and WT controls injected with MPTP/Saline as indicated ( n = 7/group). Two-way ANOVA with Tukey's post hoc test; mean +- SEM ; * p < 0.05, ** p < 0.01, *** p < 0.001, n.s., not significant

Supportive validation

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Experimental details: FIGURE 2 Neuropathological changes in VPS35 D620N KI mice. (a,e) Representative images of TH, GFAP and IBA1 immunostaining in SNpc (a) and STR (e) of 15- to 16-month-old VPS35 D620N / D620N and WT mice ( n = 6-9/group). Scale bar, 100 mum. Quantification of TH-positive neurons (b), GFAP-positive cells (c) and IBA1-positive cells (d) in SNpc. Quantification of optical density (OD) of TH-positive fibers (f), GFAP-positive cells (g), and IBA1-positive cells (h) in STR. (i-k) Representative Western blots (i) and quantification of TH (j) and GFAP (k) in VM and STR extracts from 15- to 16-month-old VPS35 D620N / D620N mice and WT controls ( n = 7/group). Student's t test, unpaired, two-tailed; data are shown as mean +- SEM ; * p < 0.05; ** p < 0.01; n.s., not significant

Supportive validation

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Experimental details: Figure 6 hMSC-EV treatment decreases colocalization between reactive astrocytes and amyloid plaques. (A) Representative immunohistochemistry images of brain slices stained with ThioS, GFAP, and DAPI. Scale bar = 1 mm. (B) We took 25 one mm 2 images throughout the brain, and Pearson's correlation coefficient (PCC) was obtained through ImageJ analysis to determine colocalization of GFAP and ThioS. **P < 0.01.

Supportive validation

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Experimental details: Fig. 3 Decreased neuronal METTL3 and m 6 A in the hippocampus by AAV-shMettl3. (A) Highly conserved gene sequences of shRNA targeting region in mouse and rat Mettl3 mRNA. (B) Knockdown effect of shMettl3 was confirmed in mouse N2a cells 3 days after transient transfection (Data are means+-SEM from 3 independent experiments). (C) AAV-mediated gene delivery in the hippocampus was confirmed by immunostaining for GFP. WT C57BL/6 mice receiving bilaterally stereotaxic injections of AAV encoding for eGFP plus anti-Mettl3 shRNA (shMettl3) or scrambled control shRNA (shCtrl) into the hippocampus at 2 months of age were analyzed 1 month later. (D-F) Representative images of the AAV-infected areas (GFP-positive) in hippocampus for different cell markers by immunocytochemistry after fixation: MAP 2 for neurons (D), GFAP for astrocytes (E), and Iba1 for microglia (F). (G, H) Representative image of immunoreactivity of METTL3 or m 6 A (G) in AAV-shRNA-injected mice and quantification analysis (H) confirmed that METTL3 protein and m 6 A modification level were reduced in mice injected with shMettl3 virus (n = 5-7 mice per group). GFP staining has been performed in adjacent brain slices at the same time and immunoreactivities of m 6 A and METTL3 were measured in GFP-positive regions. (Data are means+-SEM, * p < 0.05, **p < 0.01, (B, H) unpaired student's t-test)

Supportive validation

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Experimental details: Additional file 3: Supplementary Fig. 3. Validation of intracranial injection into hippocampus by needle track and induced neuroinflammation by METTL3 depletion in mouse hippocampus. (A) A representative image of needle track (arrow) of AAV-injected mice was shown. GFP immunoreactivity was detected in area adjacent to the needle tack. NeuN staining revealed severe neuronal loss around injected areas only in AAV-shMettl3 injected mice but not AAV-shCtrl injected mice. (B-E) Representative images of immunohistochemistry for Iba1 (B) and GFAP (Thermofisher, D) in hippocampal CA1/2 or CA3 areas in shRNA-injected mice and their quantification (C for astrocyte and E for microglia) analysis showed that METTL3 knockdown caused neuroinflammation in mouse hippocampus. (n = 4-7, *p < 0.5, **p < 0.01; C, E, unpaired student's t-test).

Supportive validation

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Experimental details: Ultrafast optogenetic inhibition in freely moving animals. A Injection, stimulation, and recordings in M1. B Activity of sorted units without (left) and with (middle) PVR management. The PVR management algorithm first subtracts an estimate of the PVR and then estimates in which trials the subtraction was incomplete. This remaining PVR trial percentage (green) is then used to weight the average across trials for each time point (lower left and right panel). The data shown are from animals 540, 548, and 549. C Relationship between opsin expression (red) and inhibition strength. To quantify the overall extracellular response, spikes were detected at a threshold of -30 uV on a per-channel basis without PVR management (PVR removed for visibility). GFAP immunostaining (green) was used to identify the BE probe location. Scale bar: 500 um. D Single-trial extracellular traces for 40 inhibitions from one electrode channel (upper panel). Histogram of spikes detected with a threshold of -30 uV (lower panel). E Latency at different percentages of the pre-inhibition spike rate bootstrapped across all sorted units. n = 212 sorted units over two animals. Boxplots: central mark indicates the median, bottom and top edges refer to the 25th and 75th percentiles of the bootstrapped data. F Illustration showing that the spike inhibition latency can be shorter than the time constant of the hyperpolarizing function. G Percentage of pre-inhibition spike rate as a function of the total light power tha

Supportive validation

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Experimental details: Figure 3 GFAP immunohistochemistry. ( A - C ) Two types of GFAP immunoreactive cells could be identified: ependymal cells (white arrowheads) and astrocytes (black arrowheads). Note the intense reactions of ECs and the processes of astrocytes that were in direct contact with the processes of the ECs.

Supportive validation

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Experimental details: Figure 5 Depletion of microglia after SCI reduces neurodegeneration in the brain sub-regions. ( A ) Experimental timeline: adult male CX3CR1-GFP mice underwent severe contusion injury at T10. Right after the surgery, PLX5622 (1200 ppm) or Vehicle chow was fed to the mice for the remainder of the study. BMS was recorded on days 1, 3, and weekly up to 6 weeks post-injury. At the 6 weeks post-injury, spinal cord tissue and brain samples were collected for IHC and histological analysis (Stereology). A subset of mice was sacrificed at 6 weeks post-injury and injured spinal cord samples were collected for Nanostring analysis. ( B ) BMS. No significant differences were found in hindlimb locomotor function on the BMS. n = 11 mice/group (2-way ANOVA with repeated measurements following Bonferroni's multiple comparisons test). ( C ) IHC representative images for GFP + cells at 0.4 mm caudal to the epicenter. Scale bar = 500 um. ( D ) IHC GFP + cell counts [n = 6 (Sham/Veh), 7 (SCI/Veh), 6 (Sham/PLX), and 5 (SCI/PLX)]. * p < 0.05, ++++ p < 0.0001, vs. Sham/Veh group. #### p < 0.0001, vs. SCI/Veh group. 2-way ANOVA following Tukey's multiple comparisons test. ( E,F ) Representative images (E) of the lesion epicenter from mice on Vehicle diet and PLX5622 diet at 6 weeks immunostained for GFAP (red) and F4/80 (blue). Scale bar =500 um. Quantification of GFAP+ area (% of total section area) from 1.4 mm rostral or caudal to the lesion epicenter (E). N = 5 mice/group. 2-way ANOVA following Si