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        Rabbit Anti-Vimentin antibody (bs-0756R)
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        说明书: 50ul  100ul  200ul
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        大包装/询价

        产品编号 bs-0756R
        英文名称 Vimentin
        中文名称 波形蛋白抗体
        别    名 FLJ36605; OTTHUMP00000019224; VIM; VIME_HUMAN; Vimentin.  
        Specific References  (61)     |     bs-0756R has been referenced in 61 publications.
        [IF=3.166] Pan et al. Roles of vimentin and 14-3-3 zeta/delta in the inhibitory effects of heparin on PC-3M cell proliferation and B16-F10-luc-G5 cells metastasis. (2012) Acta.Pharmacol.Si. 33:798-808  WB ;  Human.  
        [IF=5.008] Gao et al. Hes1 promotes cell proliferation and migration by activating Bmi-1 and PTEN/Akt/GSK3β pathway in human colon cancer. (2015) Oncotarge. 6:38667-80  WB ;  Human.  
        [IF=2.571] Tang et al. Salidroside protects against bleomycin-induced pulmonary fibrosis: activation of Nrf2-antioxidant signaling, and inhibition of NF-κB and TGF-β1/Smad-2/-3 pathways. (2016) Cell.Stress.Chaperones. 21:239-49  WB ;  Rat.  
        [IF=4.357] Fei et al. The number of polyploid giant cancer cells and epithelial-mesenchymal transition-related proteins are associated with invasion and metastasis in human breast cancer. (2015) J.Exp.Clin.Cancer.Res. 34:158  WB ;  Human.  
        [IF=1.706] Shan et al. Wnt/β-catenin pathway is required for epithelial to mesenchymal transition in CXCL12 over expressed breast cancer cells. (2016) Int.J.Clin.Exp.Pathol. 8:12357-67  WB ;  Human.  
        [IF=2.54] Chan et al. Investigating the Radioresistant Properties of Lung Cancer Stem Cells in the Context of the Tumor Microenvironment. (2016) Radiat.Res. 185:169-81  IF(ICC) ;  Human.  
        [IF=3.68] Djamali et al. Nox2 and Cyclosporine-Induced Renal Hypoxia. (2016) Transplantation. 100:1198-210  WB ;  Rat.  
        [IF=8.806] Monteiro et al. Dental cell sheet biomimetic tooth bud model. (2016) Biomaterials. 106:167-79  IHC-P,IHC-P-IF ;  Porcine.  
        [IF=4.59] Fagundes et al. Binge Drinking of Ethanol during Adolescence Induces Oxidative Damage and Morphological Changes in Salivary Glands of Female Rats. (2016) Oxid.Med.Cell.Longe. 2016:7323627  IHC-P ;  Rat.  
        [IF=4.59] Fagundes et al. Binge Drinking of Ethanol during Adolescence Induces Oxidative Damage and Morphological Changes in Salivary Glands of Female Rats. (2016) Oxid.Med.Cell.Longe. 2016:7323627  IHC-P ;  Rat.  
        [IF=4.26] Yap, Chan Choo, et al. "Different doublecortin (DCX) patient alleles show distinct phenotypes in cultured neurons: evidence for divergent loss-of-function and off-pathway cellular mechanisms." Journal of Biological Chemistry (2016): jbc-M116.  WB ;  Rat.  
        [IF=4.13] Yap et al. Different Doublecortin (DCX) Patient Alleles Show Distinct Phenotypes in Cultured Neurons: EVIDENCE FOR DIVERGENT LOSS-OF-FUNCTION AND "OFF-PATHWAY" CELLULAR MECHANISMS. (2016) J.Biol.Chem. 291:26613-26626  WB ;  Monkey, Human.  
        [IF=4.12] Wang et al. Kukoamine A inhibits human glioblastoma cell growth and migration through apoptosis induction and epithelial-mesenchymal transition attenuation. (2016) Sci.Rep. 6:36543  WB ;  Human.  
        [IF=3.83] Zhou et al. Induced Pluripotent Stem Cells Inhibit Bleomycin-Induced Pulmonary Fibrosis in Mice through Suppressing TGF-β1/Smad-Mediated Epithelial to Mesenchymal Transition. (2016) Front.Pharmacol. 7:430  WB ;  Mouse.  
        [IF=1.585] Wang et al. 5-Aminolaevulinic Acid-Based Photodynamic Therapy Restrains Pathological Hyperplasia of Fibroblasts. (2017) Med.Sci.Moni. 23:46-56  IF(ICC) ;  Human.  
        [IF=4.76] Khayat et al. GelMA-Encapsulated hDPSCs and HUVECs for Dental Pulp Regeneration. (2017) J.Dent.Res. 96:192-199  IF(ICC) ;  Human.  
        [IF=4.43] Wang et al. The Aurora-A-Twist1 axis promotes highly aggressive phenotypes in pancreatic carcinoma. (2017) J.Cell.Sci. 130:1078-1093  WB ;  Human.  
        [IF=2.81] Wang et al. Anti-proliferative activities of finasteride in benign prostate epithelial cells require stromal fibroblasts and c-Jun gene. (2017) PLoS.One. 12:e0172233  IHC-P ;  Mouse.  
        [IF=4.26] Pagani et al. Human Endometrial Stromal Cells Are Highly Permissive To Productive Infection by Zika Virus. (2017) Sci.Rep. 7:44286  IF(ICC) ;  Human.  
        [IF=4.26] Zhou et al. GDC-0449 improves the antitumor activity of nano-doxorubicin in pancreatic cancer in a fibroblast-enriched microenvironment. (2017) Sci.Rep. 7:13379  IF(IHC-F) ;  Mouse.  
        [IF=3.125] Daverey A et al. Pre and post treatment with curcumin and resveratrol protects astrocytes after oxidative stress.Brain Res. 2018 Aug 1;1692:45-55.  WB ;  Human.  
        [IF=1.438] Jiang?JY et al. Effects of estradiol and progesterone on secretion of epidermal growth factor and insulin-like growth factor-1 in cultured yak endometrial epithelial cells. Tissue Cell. 2018 Jun;52:28-34.  ICC ;  Yak.  
        [IF=3.7] Wang et al. Expression and clinical significance of autophagic protein LC3B and EMT markers in gastric cancer. (2018) Cancer.Manag.Res. 10:1479-1486  IHC ;  Human.  
        [IF=3.231] Xu W et al. An in situ hydrogel based on carboxymethyl chitosan and sodium alginate dialdehyde for corneal wound healing after alkali burn.(2018) J Biomed Mater Res A.  IHF ;  Rabbit.  
        [IF=6.491] Nikhil K et al. Identification of LIMK2 as a Therapeutic Target in Castration Resistant Prostate Cancer. Cancer Lett. 2019 Apr 28;448:182-196.  WB ;  Mouse.  
        [IF=5.646] Ye T et al. MicroRNA-7 as a potential therapeutic target for aberrant NF-κB-driven distant metastasis of gastric cancer. J Exp Clin?Cancer?Res.?2019 Feb 6;38(1):55.  FCM&IHC ;  Mouse&Human.  
        [IF=2.852] Tarique I et al. LIPOPHAGY: a novel form of steroidogenic activity within the LEYDIG cell during the reproductive cycle of turtle. Reprod Biol Endocrinol. 2019 Feb 9;17(1):19.   IHC-P&IF ;  Pelodiscus sinensis.  
        [IF=2.705] Zhou S et al. Helicobacter?pylori?infection?promotes?epithelial-to-mesenchymal?transition?of?gastric?cells?by?upregulating?LAPTM4B. Biochem Biophys Res Commun. 2019 Jun 30;514(3):893-900.  IHC-P,ICF&WB ;  Human.  
        [IF=3.559] Yin X et al. Zinc?oxide?nanoparticles?ameliorate?collagen?lattice?contraction?in?human?tenon?fibroblasts. Arch Biochem Biophys.?2019 Jul 15;669:1-10.?  ICC ;  Human.  
        [IF=0] Khuder, Saja S. Regulation of Expression of CEACAM1 and Functional Correlation in Metabolic Diseases. Diss. University of Toledo, 2016.  IHC-P ;  Mouse.  
        [IF=2.17] Li T et al. Tissue-engineered scaffold based on carboxymethyl chitin or chitosan for corneal epithelial transplantation. Polymer Journal,2018 50(7), 511–521.   IF&WB ;  Rabbit.  
        [IF=3.32] Hu, Bin, et al. "IFN‐γ Inhibits Osteopontin Expression in Human Decidual Stromal Cells and can be Attenuated by 1α, 25‐Dihydroxyvitamin D3." American Journal of Reproductive Immunology 68.4 (2012): 353-361.  Human.  
        [IF=5.01] Wang, Li-Ping, et al. "Angiotensin II upregulates K Ca 3.1 channels and stimulates cell proliferation in rat cardiac fibroblasts." Biochemical pharmacology 85.10 (2013): 1486-1494.  Rat.  
        [IF=2.41] Liu, Yang, et al. "Down-regulation of< i> Wnt10a affects odontogenesis and proliferation in mesenchymal cells." Biochemical and Biophysical Research Communications (2013).  Mouse.  
        [IF=3.73] Wang, Yueqin, et al. "Hypoxia-Inducible Factor-1alpha and MAPK Co-Regulate Activation of Hepatic Stellate Cells upon Hypoxia Stimulation."?PLoS ONE?8.9 (2013).  WB ;  Mouse.  
        [IF=.96] Fu, Min, et al. "Effects of Ureaplasma urealyticum lipid-associated membrane proteins on rheumatoid arthritis synovial fibroblasts." Journal of International Medical Research 41.5 (2013): 1655-1670.  Human.  
        [IF=3.44] Li, Yingli, et al. "Differences between Niche Cells and Limbal Stromal Cells in Maintenance of Corneal Limbal Stem Cells." Investigative Ophthalmology & Visual Science (2014): IOVS-13  Human.  
        [IF=3.98] Li, Junqin, et al. "Fatty acid synthase mediates the epithelial-mesenchymal transition of breast cancer cells." International Journal of Biological Sciences10.2 (2014): 171.  WB ;  Human.  
        [IF=2.06] Wang, Jing, et al. "Immortalized porcine intestinal epithelial cell cultures susceptible to porcine rotavirus infection." Journal of Virological Methods (2014).  Pig.  
        [IF=3.06] Sottnik, Joseph L., et al. "Osteocytes Serve as a Progenitor Cell of Osteosarcoma." Journal of Cellular Biochemistry (2014).  IHC-P ;  Mouse.  
        [IF=4.12] Pan, Yan, et al. "Enoxaparin Sensitizes Human Non-small-cell Lung Carcinomas to Gefitinib by Inhibiting DOCK1 Expression, Vimentin Phosphorylation and Akt Activation." Molecular pharmacology (2014): mol-114.  WB ;  Human.  
        [IF=2.38] Tong, Weihua, et al. "Sorcin Enhances Metastasis and Promotes Epithelial-to-Mesenchymal Transition of Colorectal Cancer." Cell Biochemistry and Biophysics (2015): 1-7.  WB ;  Human.  
        [IF=2.38] Cheng, Gangwei, et al. "Direct Effects of Bevacizumab on Rat Conjunctival Fibroblast." Cell Biochemistry and Biophysics: 1-6.  Rat.  
        [IF=2.84] Deng, Boya, et al. "MicroRNA-142-3p inhibits cell proliferation and invasion of cervical cancer cells by targeting FZD7." Tumor Biology (2015): 1-9.  WB ;  Human.  
        [IF=3.38] Fernández-Ferreiro, Anxo, et al. "In vitro and in vivo ocular safety and eye surface permanence determination by direct and Magnetic Resonance Imaging of ion-sensitive hydrogels based on Gellan gum and kappa-carrageenan."European Journal of Pharmaceutics and Biopharmaceutics (2015).  IHC-F ;  Rat.  
        [IF=5.68] Karim, A. S., et al. "Nox2 Is a Mediator of Ischemia Reperfusion Injury."American Journal of Transplantation (2015).  WB ;  Mouse.  
        [IF=1.89] Su, Jing, and Hong Li. "RAC1 overexpression promotes the proliferation, migration and epithelial-mesenchymal transition of lens epithelial cells." International Journal of Clinical and Experimental Pathology 8.9 (2015): 10760-10767.  WB ;  Human.  
        [IF=1.28] Chen, Jia, et al. "Biological characterization of metanephric mesenchymal stem cells from the Beijing duck." Experimental and Therapeutic Medicine 11.2 (2016): 439-447.  FCM ;  Others.  
        [IF=4.72] Sucre, Jennifer MS, et al. "A three-dimensional human model of the fibroblast activation that accompanies bronchopulmonary dysplasia identifies Notch-mediated pathophysiology." American Journal of Physiology-Lung Cellular and Molecular Physiology 310.10 (2016): L889-L898.  IF(IHC-P) ;  Human.  
        [IF=1.7] Huang, Chao, and Bin Wen. "Phenotype transformation of immortalized NCM460 colon epithelial cell line by TGF-β1 is associated with chromosome instability." Molecular Biology Reports 43.10 (2016): 1069-1078.  IF(ICC) ;  Human.  
        [IF=3.23] Daverey, Amita, and Sandeep K. Agrawal. "Curcumin alleviates oxidative stress and mitochondrial dysfunction in astrocytes." Neuroscience 333 (2016): 92-103.  WB ;  Human.  
        [IF=8.39] Monteiro, Nelson, et al. "Dental Cell Sheet Biomimetic Tooth Bud Model."Biomaterials (2016).  IF(IHC-P) ;  Pig.  
        [IF=5.03] Wilkinson, Dan C., et al. "Development of a Three-Dimensional Bioengineering Technology to Generate Lung Tissue for Personalized Disease Modeling." Stem Cells Translational Medicine (2016): sctm-2016.  IHC-P ;  Human.  
        [IF=1.46] Xiao, Jiajia, et al. "Isolation of Bovine Skin-Derived Precursor Cells and Their Developmental Potential After Nuclear Transfer." Cellular Reprogramming (Formerly" Cloning and Stem Cells") 18.6 (2016): 411-418.  IF ;  Bovine.  
        [IF=4.71] Smith, Elizabeth E., et al. "Developing a biomimetic tooth bud model." Journal of Tissue Engineering and Regenerative Medicine (2017).  IF(IHC-P) ;  Human.  
        [IF=7.43] Wang, Jing, et al. "Phosphorylation-dependent regulation of ALDH1A1 by Aurora kinase A: insights on their synergistic relationship in pancreatic cancer." BMC biology 15.1 (2017): 10.  WB ;  Human.  
        [IF=2.81] Tanos, T., et al. "Isolation of putative stem cells present in human adult olfactory mucosa." PloS one 12.7 (2017): e0181151.  IHC-P ;  Human.  
        [IF=1.84] Park, Song, et al. "Critical roles of hMAGEA2 in induced pluripotent stem cell pluripotency, proliferation, and differentiation." Cell Biochemistry and Function (2017).  IHC-P ;  Human.  
        [IF=3.046] Qiao J et al. Knockdown of ROS proto-oncogene 1 inhibits migration and invasion in gastric cancer cells by targeting the PI3K/Akt signaling pathway. OncoTargets and Therapy, 2019,12: 8569–8582.   WB ;  Human.  
        [IF=2.336] Wang S et al. Micro RNA‐198‐5p inhibits the migration and invasion of non‐small lung cancer cells by targeting fucosyltransferase 8. Clinical and Experimental Pharmacology and Physiology. 2019.  ICF ;  Human.  
        [IF=4.627] Jin L et al. Mesenchymal stem cells ameliorate myocardial fibrosis in diabetic cardiomyopathy via the secretion of prostaglandin E2. Stem Cell Res Ther. 2020 Mar 17;11(1):122.  IF ;  rat.  
        研究领域 肿瘤  细胞生物  免疫学  发育生物学  神经生物学  信号转导  干细胞  细胞骨架  肿瘤细胞生物标志物  
        抗体来源 Rabbit
        克隆类型 Polyclonal
        交叉反应 Human, Mouse,  (predicted: Rat, Chicken, Pig, Cow, Goat, )
        产品应用 WB=1:500-2000 ELISA=1:5000-10000 IHC-P=1:100-500 IHC-F=1:100-500 Flow-Cyt=1μg/Test ICC=1:100-500 IF=1:100-500 (石蜡切片需做抗原修复)
        not yet tested in other applications.
        optimal dilutions/concentrations should be determined by the end user.
        分 子 量 53kDa
        细胞定位 细胞浆 细胞外基质 
        性    状 Liquid
        浓    度 1mg/ml
        免 疫 原 KLH conjugated synthetic peptide derived from human Vimentin:371-466/466 
        亚    型 IgG
        纯化方法 affinity purified by Protein A
        储 存 液 0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol.
        保存条件 Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.
        PubMed PubMed
        产品介绍 This gene encodes a member of the intermediate filament family. Intermediate filamentents, along with microtubules and actin microfilaments, make up the cytoskeleton. The protein encoded by this gene is responsible for maintaining cell shape, integrity of the cytoplasm, and stabilizing cytoskeletal interactions. It is also involved in the immune response, and controls the transport of low-density lipoprotein (LDL)-derived cholesterol from a lysosome to the site of esterification. It functions as an organizer of a number of critical proteins involved in attachment, migration, and cell signaling. Mutations in this gene causes a dominant, pulverulent cataract.[provided by RefSeq, Jun 2009]

        Function:
        Vimentins are class-III intermediate filaments found in various non-epithelial cells, especially mesenchymal cells. Vimentin is attached to the nucleus, endoplasmic reticulum, and mitochondria, either laterally or terminally.
        Involved with LARP6 in the stabilization of type I collagen mRNAs for CO1A1 and CO1A2.

        Subunit:
        Homopolymer assembled from elementary dimers. Interacts with HCV core protein. Interacts with LGSN and SYNM. Interacts (via rod region) with PLEC (via CH 1 domain) (By similarity). Interacts with SLC6A4. Interacts with STK33. Interacts with LARP6. Interacts with RAB8B (By similarity).

        Subcellular Location:
        Cytoplasm.

        Tissue Specificity:
        Highly expressed in fibroblasts, some expression in T- and B-lymphocytes, and little or no expression in Burkitt's lymphoma cell lines. Expressed in many hormone-independent mammary carcinoma cell lines.

        Post-translational modifications:
        Filament disassembly during mitosis is promoted by phosphorylation at Ser-55 as well as by nestin (By similarity). One of the most prominent phosphoproteins in various cells of mesenchymal origin. Phosphorylation is enhanced during cell division, at which time vimentin filaments are significantly reorganized. Phosphorylation by PKN1 inhibits the formation of filaments. Phosphorylated at Ser-56 by CDK5 during neutrophil secretion in the cytoplasm. Phosphorylated by STK33.

        Similarity:
        Belongs to the intermediate filament family.

        SWISS:
        P08670

        Gene ID:
        7431

        Database links:

        Entrez Gene: 7431 Human

        Entrez Gene: 22352 Mouse

        Entrez Gene: 81818 Rat

        Omim: 193060 Human

        SwissProt: P08670 Human

        SwissProt: P20152 Mouse

        SwissProt: P31000 Rat

        Unigene: 455493 Human



        Important Note:
        This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.

        Vimentin—波形蛋白。是五种主要的中间丝之一,存在于各种正常和病理性间质来源的组织,如纤维母细胞、内皮细胞、淋巴细胞等正常细胞和肉瘤、淋巴瘤、黑色素瘤等肿瘤。波形蛋白是负责维持细胞骨架完整性的蛋白之一。
        产品图片
        Sample:
        Hela(Human) Cell Lysate at 30 ug
        Hela KO Vimentin (Human) Cell Lysate at 30 ug
        Primary: Anti- Vimentin (bs-0756R) at 1/1000 dilution
        Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
        Predicted band size: 53 kD
        Observed band size: 53 kD
        Sample:
        Cerebrum (Mouse) Lysate at 40 ug
        Primary: Anti-Vimentin (bs-0756R) at 1/2000 dilution
        Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
        Predicted band size: 53 kD
        Observed band size: 53 kD
        Sample: Hela Cell (Human) Lysate at 40 ug
        Primary: Anti-Vimentin (bs-0756R) at 1/300 dilution
        Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
        Predicted band size: 53 kD
        Observed band size: 53 kD
        Sample: Jurkat (human)Cell Lysate at 40 ug
        Primary: Anti- Vimentin (bs-0756R) at 1/300 dilution
        Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
        Predicted band size: 53 kD
        Observed band size: 53 kD
        Sample: Hela (human)Cell Lysate at 40 ug
        Primary: Anti- Vimentin (bs-0756R) at 1/300 dilution
        Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
        Predicted band size: 53 kD
        Observed band size: 53 kD
        Sample:
        Lane 1: Hela (Human) Cell Lysate at 30 ug
        Lane 2: NIH/3T3 (Mouse) Cell Lysate at 30 ug
        Lane 3: U251 (Human) Cell Lysate at 30 ug
        Lane 4: SH-SY5Y (Human) Cell Lysate at 30 ug
        Lane 5: A549 (Human) Cell Lysate at 30 ug
        Primary: Anti-Vimentin (bs-0756R) at 1/1000 dilution
        Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
        Predicted band size: 55 kD
        Observed band size: 57 kD
        Paraformaldehyde-fixed, paraffin embedded (human cervical cancer); 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 (Vimentin) Polyclonal Antibody, Unconjugated (bs-0756R) at 1:400 overnight at 4°C, followed by a conjugated secondary (sp-0023) for 20 minutes and DAB staining.
        Paraformaldehyde-fixed, paraffin embedded (human cervical cancer); 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 (Vimentin) Polyclonal Antibody, Unconjugated (bs-0756R) at 1:400 overnight at 4°C, followed by a conjugated secondary (sp-0023) for 20 minutes and DAB staining.
        Tissue/cell: mouse kidney 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-Vimentin Polyclonal Antibody, Unconjugated(bs-0756R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
        Tissue/cell: HeLa 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 (Vimentin) Polyclonal Antibody, Unconjugated (bs-0756R) 1:50, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody (bs-0295G-FITC) at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.
        Tissue/cell: U251 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 (Vimentin) Polyclonal Antibody, Unconjugated (bs-0756R) 1:50, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody (bs-0295G-FITC) at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.
        Tissue/cell: U-87MG 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 (Vimentin) Polyclonal Antibody, Unconjugated (bs-0756R) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody (bs-0295G-FITC) at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.
        Tissue/cell: U-2OS 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 (Vimentin) Polyclonal Antibody, Unconjugated (bs-0756R) 1:50, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody (bs-0295G-FITC) at 37°C for 90 minutes, DAPI (5ug/ml, blue, C-0033) was used to stain the cell nuclei.
        Tissue/cell: mouse mesenchymal stem cells;
        Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
        Incubation: Anti-Vimentin Polyclonal Antibody, Unconjugated(bs-0756R) 1:300, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
        Blank control: Jurkat cells(blue).
        Primary Antibody:Rabbit Anti-Vimentin antibody antibody(bs-0756R), 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-0756R, 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.
        Blank control:A549.
        Primary Antibody (green line): Rabbit Anti-Vimentin antibody (bs-0756R)
        Dilution: 1μg /10^6 cells;
        Isotype Control Antibody (orange line): Rabbit IgG .
        Secondary Antibody : Goat anti-rabbit IgG-AF488
        Dilution: 1μg /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.
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