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          RabbitAnti-GFAP antibody (bs-0199R)
          ~~~促銷,代碼KXJ230206~~~
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          產品編號 bs-0199R
          英文名稱 GFAP
          中文名稱 膠質纖維酸性蛋白抗體
          別    名 Astrocyte Marker; FLJ45472; GFAP; Glial Fibrillary Acidic Protein; Intermediate filament protein; GFAP_HUMAN.  
          Specific References  (74)     |     bs-0199R has been referenced in 74 publications.
          111 [IF=10.82] Zhao, Hongyu, et al. "Mice deficient in Epg5 exhibit selective neuronal vulnerability to degeneration." The Journal of Cell Biology (2013).  IHC-P ;  Mouse.  222
          111 [IF=4.65] Zhao, Yan G., et al. "The p53-induced gene Ei24 is an essential component of the basal autophagy pathway." Journal of Biological Chemistry 287.50 (2012): 42053-42063.  Mouse.  222
          111 [IF=3.73] Xiang, Yanxiao, et al. "Anti-inflammatory Effect of Acetylpuerarin on Eicosanoid Signaling Pathway in Primary Rat Astrocytes."Journal of Molecular Neuroscience(2013): 1-9  IF(ICC) ;  Rat.  222
          111 [IF=2.93] Liu, Yang, et al. "A Simple Method for Isolating and Culturing the Rat Brain Microvascular Endothelial Cells." Microvascular Research (2013).  Rat.  222
          111 [IF=2.89] Xiang, Yanxiao, et al. "Anti-inflammatory Effect of Acetylpuerarin on Eicosanoid Signaling Pathway in Primary Rat Astrocytes."Journal of Molecular Neuroscience(2013): 1-9  Mouse.  222
          111 [IF=1.29] Fan, Lixing, et al. "Directed differentiation of aged human bone marrow multipotent stem cells effectively generates dopamine neurons."?In Vitro Cellular & Developmental Biology-Animal?(2013): 1-9.  Human.  222
          111 [IF=2.65] Zuo, Daiying, et al. "Existence of glia mitigated ketamine-induced neurotoxicity in neuron-glia mixed cultures of neonatal rat cortex and the glia-mediated protective effect of 2-PMPA." Neurotoxicology (2014).  Rat.  222
          111 [IF=10.53] Ma, Benyu, et al. "Dapper1 promotes autophagy by enhancing the Beclin1-Vps34-Atg14L complex formation." Cell Research (2014).  IHC-F ;  Mouse.  222
          111 [IF=7.58] Shan, Chun-Lei, et al. "High Efficiency Intracellular Transport of Cationic Peptide Stearate for Gene Delivery in Tumor Cells and Multipotent Stem Cells." Journal of Biomedical Nanotechnology 10.11 (2014): 3231-3243.  other ;  
          111 [IF=5.29] Du, Wenzhong, et al. "Targeting the SMO oncogene by miR-326 inhibits glioma biological behaviors and stemness." Neuro-Oncology (2014): nou217.  IHC-F ;  Human.  222
          111 [IF=11.42] Zhao, Yan G., et al. "The autophagy gene Wdr45/Wipi4 regulates learning and memory function and axonal homeostasis." Autophagy (2015).  IHC-P ;  Mouse.  222
          111 [IF=2.86] Mori, Miki, et al. "Stromal Cell-Derived Factor-1α Plays a Crucial Role Based on Neuroprotective Role in Neonatal Brain Injury in Rats." International Journal of Molecular Sciences 16.8 (2015): 18018-18032.  IHC-F ;  Rat.  222
          111 [IF=2.47] Yan, Yu-hui, et al. "Osthole Protects Bone Marrow-Derived Neural Stem Cells from Oxidative Damage through PI3K/Akt-1 Pathway." Neurochemical Research (2016): 1-8.  other ;  Mouse.  222
          111 [IF=1.812] Liao, Wei-Tao, et al. "The effect of celastrol on learning and memory in diabetic rats after sevoflurane inhalation." (2016).  IHC-P ;  Rat.  222
          111 [IF=2.86] Hu. et al. Levo-Corydalmine Alleviates Neuropathic Cancer Pain Induced by Tumor Compression via the CCL2/CCR2 Pathway  IF(IHC-P) ;  Mouse.  222
          111 [IF=1.77] Liang et al. Scorpion ethanol extract and valproic acid effects on hippocampal glial fibrillary acidic protein expression in a rat model of chronic-kindling epilepsy induced by lithium chloride-pilocarpine. (2012) Neural.Regen.Re. 7:426-33  IHC ;  rat.  222
          111 [IF=1.97] Liao et al. The effect of celastrol on learning and memory in diabetic rats after sevoflurane inhalation. (2018) Arch.Med.Sci. 14:370-380  IHC ;  Rat.  222
          111 [IF=3.974] Esaki,et al.ASC amino acid transporter 2, defined by enzyme-mediated activation of radical sources, enhances malignancy of GD2-positive small-cell lung cancer.(2018) Cancer Science. 109:141-153.  IF(ICC) ;  Human.  222
          111 [IF=3.562] Zhang,et al.Interferon-γ Promotes Neuronal Repair by Transplanted Neural Stem Cells in Ischemic Rats.(2018) Stem Cells and Development. 27:355-366.  IF(ICC) ;  Rat.  222
          111 [IF=2.341] Zhong,et al.Protective effect of adenovirus-mediated erythropoietin expression on the spiral ganglion neurons in the rat inner ear.(2018) International Journal of Molecular Medicine. 41:2669-2677.  IHC-P + IF(IHC-P) ;  Rat.  222
          111 [IF=2.81] Farah et al. Tau accumulations in the brains of woodpeckers. (2018) PLoS.One. 13:e0191526  IHC-P ;  Woodpecker.  222
          111 [IF=2.234] Lu et al. Autophagy activator promotes neuronal differentiation of adult adipose-derived stromal cells. (2013) Neural.Regen.Res. 8:882-9  ICC ;  Human.  222
          111 [IF=0] ?Yang?JT et al. Sex Differences in Neuropathology and Cognitive Behavior in APP/PS1/tau Triple-Transgenic Mouse Model of Alzheimer's Disease. Neurosci Bull. 2018 Oct;34(5):736-746.   IHC ;  Mouse.  222
          111 [IF=3.44] Zhang X et al.Neuroprotective effects of matrix metalloproteinases in cerebral ischemic rats by promoting activation and migration of astrocytes and microglia.(2018) Brain Res Bull. Nov 13.  WB ;  Rat.  222
          111 [IF=2.064] Yao Q et al.Lycium Barbarum Polysaccharides Improve Retinopathy in Diabetic Sprague-Dawley Rats.(2018)Evid Based Complement Alternat Med.Nov 15;2018:7943212.  WB ;  Rat.  222
          111 [IF=2.766] Xiang Y et al. Inhibition of sPLA. sub. 2-IIA Prevents LPS-Induced Neuroinflammation by Suppressing ERK1/2-cPLA. sub. 2 [alpha] Pathway in Mice Cerebral Cortex.PLoS One. 2013 Oct 9;8(10):e77909.  Other ;  
          111 [IF=2.159] Iwasa T et al. IL-10 and CXCL2 in trigeminal ganglia in neuropathic pain.Neurosci Lett.?2019 Jun 11;703:132-138.  IHC-P ;  Rat.  222
          111 [IF=3.811] Sun Z et al. Glioblastoma Stem Cell-Derived Exosomes Enhance Stemness and Tumorigenicity of Glioma Cells by Transferring Notch1 Protein. Cell Mol Neurobiol. 2019 Dec 18.  WB ;  Human&Mouse.  222
          111 [IF=.181] H Mu et al. HCMV-encoded IE2 induces anxiety-depression and cognitive impairment in UL122 genetically-modified mice. Int J Clin Exp Pathol 2019;12(11):4087-4095.  WB&IHC-P ;  Mouse.  222
          111 [IF=3.471] Chen J et al. Modulation of activated astrocytes in the hypothalamus paraventricular nucleus to prevent ventricular arrhythmia complicating acute myocardial infarction. Int J Cardiol. 2020 Jan 16.  WB&IHC-P ;  Rat.  222
          研究領域 腫瘤  細胞生物  免疫學  神經生物學  信號轉導  干細胞  細胞粘附分子  細胞類型標志物  細胞骨架  
          抗體來源 Rabbit
          克隆類型 Polyclonal
          交叉反應 Human, Mouse, Rat,  (predicted: Dog, Pig, Cow, Rabbit, Sheep, )
          產品應用 WB=1:500-2000 ELISA=1:5000-10000 IHC-P=1:200-1000 IHC-F=1:200-1000 Flow-Cyt=1μg/Test IF=1:200-800 (石蠟切片需做抗原修復)
          not yet tested in other applications.
          optimal dilutions/concentrations should be determined by the end user.
          理論分子量 48kDa
          細胞定位 細胞漿 
          性    狀 Liquid
          濃    度 1mg/ml
          免 疫 原 KLH conjugated synthetic peptide derived from human GFAP: 51-150/432 
          亞    型 IgG
          純化方法 affinity purified by Protein A
          緩 沖 液 0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol.
          保存條件 Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.
          注意事項 This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.
          PubMed PubMed
          產品介紹 This gene encodes one of the major intermediate filament proteins of mature astrocytes. It is used as a marker to distinguish astrocytes from other glial cells during development. Mutations in this gene cause Alexander disease, a rare disorder of astrocytes in the central nervous system. Alternative splicing results in multiple transcript variants encoding distinct isoforms. [provided by RefSeq, Oct 2008]

          Function:
          GFAP, a class-III intermediate filament, is a cell-specific marker that, during the development of the central nervous system, distinguishes astrocytes from other glial cells.

          Subunit:
          Interacts with SYNM. Isoform 3 interacts with PSEN1 (via N-terminus).

          Subcellular Location:
          Cytoplasm. Note=Associated with intermediate filaments.

          Tissue Specificity:
          Expressed in cells lacking fibronectin.

          Post-translational modifications:
          Phosphorylated by PKN1.

          DISEASE:
          Defects in GFAP are a cause of Alexander disease (ALEXD) [MIM:203450]. Alexander disease is a rare disorder of the central nervous system. It is a progressive leukoencephalopathy whose hallmark is the widespread accumulation of Rosenthal fibers which are cytoplasmic inclusions in astrocytes. The most common form affects infants and young children, and is characterized by progressive failure of central myelination, usually leading to death usually within the first decade. Infants with Alexander disease develop a leukoencephalopathy with macrocephaly, seizures, and psychomotor retardation. Patients with juvenile or adult forms typically experience ataxia, bulbar signs and spasticity, and a more slowly progressive course.

          Similarity:
          Belongs to the intermediate filament family.

          SWISS:
          P14136

          Gene ID:
          2670

          Database links:

          Entrez Gene: 281189 Cow

          Entrez Gene: 2670 Human

          Entrez Gene: 14580 Mouse

          Entrez Gene: 24387 Rat

          Omim: 137780 Human

          SwissProt: Q28115 Cow

          SwissProt: P14136 Human

          SwissProt: P03995 Mouse



          星形膠質細胞標志物 (Astrocyte Marker)

          GFAP是一個56kDa的中間絲蛋白(intermediate filament,IF),在中樞神經系統發育期是一個特異性的標志物,以區別星形細胞和其它膠質細胞。GFAP表達在皮層和海馬,急、慢性皮質酮治療時表達減少。
          GFAP可以和人、大鼠、小鼠的GFAP反應,在正常和腫瘤性的星形膠質細胞陽性表達,而神經節細胞、神經元、成纖維細胞、少突膠質細胞和這些細胞來源的腫瘤細胞陰性表達,主要用于星形膠質瘤等中樞神經系統腫瘤的診斷和鑒別診斷,GFAP的缺乏可導致AD病。
          產品圖片
          Sample:Optic nerve (Rat)cell Lysate at 40 ug
          Primary: Anti-GFAP(bs-0199R)at 1/300 dilution
          Secondary: IRDye800CW Goat Anti-RabbitIgG at 1/20000 dilution
          Predicted band size: 48 kD
          Observed band size: 53 kD
          Sample: U251 Cell Lysate at 40 ug
          Primary: Anti- GFAP (bs-0199R) at 1/300 dilution
          Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/10000 dilution
          Predicted band size: 48 kD
          Observed band size: 50 kD
          Sample:
          Cerebellum (Rat) Lysate at 40 ug
          Cerebellum (Mouse) Lysate at 40 ug
          Eye (Mouse) Lysate at 40 ug
          Primary: Anti-GFAP (bs-0199R) at 1/300 dilution
          Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
          Predicted band size: 48 kD
          Observed band size: 48 kD
          Paraformaldehyde-fixed, paraffin embedded (human glioma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (GFAP) Polyclonal Antibody, Unconjugated (bs-0199R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
          Tissue/cell: rat brain tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
          Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
          Incubation: Anti-GFAP Polyclonal Antibody, Unconjugated(bs-0199R) 1:400, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
          Paraformaldehyde-fixed, paraffin embedded (rat cerebellum); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (GFAP) Polyclonal Antibody, Unconjugated (bs-0199R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
          Paraformaldehyde-fixed, paraffin embedded (mouse cerebellum); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (GFAP) Polyclonal Antibody, Unconjugated (bs-0199R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
          Paraformaldehyde-fixed, paraffin embedded (rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (GFAP) Polyclonal Antibody, Unconjugated (bs-0199R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
          Paraformaldehyde-fixed, paraffin embedded (mouse brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (GFAP) Polyclonal Antibody, Unconjugated (bs-0199R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
          Paraformaldehyde-fixed, paraffin embedded (mouse brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (GFAP) Polyclonal Antibody, Unconjugated (bs-0199R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
          Paraformaldehyde-fixed, paraffin embedded (rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (GFAP) Polyclonal Antibody, Unconjugated (bs-0199R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
          Paraformaldehyde-fixed, paraffin embedded (human brain ); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (GFAP) Polyclonal Antibody, Unconjugated (bs-0199R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
          Paraformaldehyde-fixed, paraffin embedded (human brain glioma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (GFAP) Polyclonal Antibody, Unconjugated (bs-0199R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
          Paraformaldehyde-fixed, paraffin embedded (mouse brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (GFAP) Polyclonal Antibody, Unconjugated (bs-0199R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
          Paraformaldehyde-fixed, paraffin embedded (mouse cerebellum); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (GFAP) Polyclonal Antibody, Unconjugated (bs-0199R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
          Paraformaldehyde-fixed, paraffin embedded (rat cerebellum); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (GFAP) Polyclonal Antibody, Unconjugated (bs-0199R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
          Paraformaldehyde-fixed, paraffin embedded (mouse brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (GFAP) Polyclonal Antibody, Unconjugated (bs-0199R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
          Paraformaldehyde-fixed, paraffin embedded (rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (GFAP) Polyclonal Antibody, Unconjugated (bs-0199R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
          Paraformaldehyde-fixed, paraffin embedded (Mouse brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (GFAP) Polyclonal Antibody, Unconjugated (bs-0199R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
          Tissue/cell: rat brain tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
          Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
          Incubation: Anti-GFAP Polyclonal Antibody, Unconjugated(bs-0199R) 1:500, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
          Tissue/cell:U-251 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37°C for 20 min; Antibody incubation with (GFAP) polyclonal Antibody, Unconjugated (bs-0199R) 1:100, 90 minutes at 37°C; followed by a FITC conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.
          U87MG cell; 4% Paraformaldehyde-fixed; Ice-cold methanol at -20℃ for 20 min; Blocking buffer (normal goat serum, C-0005) at 37°C for 20 min; Antibody incubation with (GFAP) polyclonal Antibody, Unconjugated (bs-0199R) 1:100, 90 minutes at 37°C; followed by a FITC conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.
          Paraformaldehyde-fixed, paraffin embedded (rat cerebellum); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (GFAP) Polyclonal Antibody, Unconjugated (bs-0199R) at 1:300 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nuclei staining.
          Paraformaldehyde-fixed, paraffin embedded (human brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (GFAP) Polyclonal Antibody, Unconjugated (bs-0199R) at 1:500 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nuclei staining.
          Paraformaldehyde-fixed, paraffin embedded (rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (GFAP) Polyclonal Antibody, Unconjugated (bs-0199R) at 1:500 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nuclei staining.
          Paraformaldehyde-fixed, paraffin embedded (mouse cerebellum); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (GFAP) Polyclonal Antibody, Unconjugated (bs-0199R) at 1:500 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nuclei staining.
          Paraformaldehyde-fixed, paraffin embedded (mouse brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (GFAP) Polyclonal Antibody, Unconjugated (bs-0199R) at 1:500 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nuclei staining.
          Paraformaldehyde-fixed, paraffin embedded (mouse cerebellum); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (GFAP) Polyclonal Antibody, Unconjugated (bs-0199R) at 1:200 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (bs-0295G-FITC) for 90 minutes, and DAPI for nuclei staining.
          Paraformaldehyde-fixed, paraffin embedded (rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (GFAP) Polyclonal Antibody, Unconjugated (bs-0199R) at 1:200 overnight at 4°C, followed by a conjugated secondary (bs-0295G-Cy3) at [1:500] for 90 minutes and DAPI staining of the nuclei.
          Tissue/cell: rat brain tissue;4% Paraformaldehyde-fixed and paraffin-embedded;
          Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
          Incubation: Anti-GFAP Polyclonal Antibody, Unconjugated(bs-0199R) 1:400, overnight at 4°C; The secondary antibody was Goat Anti-Rabbit IgG, Cy3 conjugated(bs-0295G-Cy3)used at 1:200 dilution for 40 minutes at 37°C. DAPI(5ug/ml,blue,C-0033) was used to stain the cell nuclei
          Blank control:U87MG.
          Primary Antibody (green line): Rabbit Anti-GFAP antibody (bs-0199R)
          Dilution: 1ug/Test;
          Secondary Antibody : Goat anti-rabbit IgG-FITC
          Dilution: 0.5ug/Test.
          Protocol
          The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at -20℃.The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
          Blank control: RSC96(blue).
          Primary Antibody:Rabbit Anti- GFAP antibody(bs-0199R), Dilution: 1μg in 100 μL 1X PBS containing 0.5% BSA;
          Isotype Control Antibody: Rabbit IgG(orange) ,used under the same conditions );
          Secondary Antibody: Goat anti-rabbit IgG-PE(white blue), Dilution: 1:200 in 1 X PBS containing 0.5% BSA.
          Protocol
          The cells were fixed with 2% paraformaldehyde (10 min) , then permeabilized with 90% ice-cold methanol for 30 min on ice. Primary antibody (bs-0199R, 1μg /1x10^6 cells) were incubated for 30 min on the ice, followed by 1 X PBS containing 0.5% BSA + 1 0% goat serum (15 min) to block non-specific protein-protein interactions. Then the Goat Anti-rabbit IgG/PE antibody was added into the blocking buffer mentioned above to react with the primary antibody at 1/200 dilution for 30 min on ice. Acquisition of 20,000 events was performed.
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