Primary antibodies used for immunoblotting, co-immunofluorescence (co-IF), and immunohistochemistry: ARID1A/BAF250A (Cell Signaling Technologies, D2A8U), VIMENTIN (Cell Signaling Technologies, D21H3) for co-IF with E-CADHERIN and immunohistochemistry, E-CADHERIN (Cell Signaling Technologies, 4A2), Cytokeratin 19 (Santa Cruz Biotechnologies, A-3), KI-67 (MIB-1) (Dako), Cleaved CASPASE 3 (A175) (Cell Signaling Technologies, 5A1E), HSP90 alpha (Invitrogen, PA3–013), VIMENTIN (Cell Signaling Technologies, 5G3F10) for immunoblotting and co-IF with HSP90. ARID1B/BAF250B (Cell Signaling Technologies, E9J4T), PBRM1/BAF180 (Cell Signaling Technologies, D3F7O), BRM (Cell Signaling Technologies, D9E8B), BRG1 (Cell Signaling Technologies, A52), SMARCC1/BAF155 (Cell Signaling Technologies, D7F8S), SMARCC2/BAF170 (Cell Signaling Technologies, D8O9V), BAF60a (Santa Cruz Biotechnologies, 23), BAF53 (Santa Cruz Biotechnologies, C-7), SMARCB1/BAF47 (Cell Signaling Technologies, D8M1X). Chemical reagents: NVP-AUY-922 (LC Laboratories).
Vimentin
Manufactured by Cell Signaling Technology
Sourced in United States, United Kingdom, China, France, Denmark, Macao
Vimentin is a protein commonly used as a marker for mesenchymal cells and their derivatives. It is a type III intermediate filament that is important for maintaining cell structure and integrity.
Automatically generated - may contain errors
Lab products found in correlation
E cadherin, by Cell Signaling Technology (933 mentions)
N cadherin, by Cell Signaling Technology (571 mentions)
Snail, by Cell Signaling Technology (255 mentions)
β actin, by Cell Signaling Technology (217 mentions)
Gapdh, by Cell Signaling Technology (205 mentions)
β catenin, by Cell Signaling Technology (195 mentions)
Pvdf membrane, by Merck Group (128 mentions)
P akt, by Cell Signaling Technology (115 mentions)
Mmp 9, by Cell Signaling Technology (85 mentions)
Cleaved caspase 3, by Cell Signaling Technology (70 mentions)
β actin, by Santa Cruz Biotechnology (67 mentions)
β actin, by Merck Group (66 mentions)
Mmp 2, by Cell Signaling Technology (62 mentions)
Bcl 2, by Cell Signaling Technology (61 mentions)
E cadherin, by BD (60 mentions)
Gapdh, by Santa Cruz Biotechnology (57 mentions)
Bcl2 associated x (bax), by Cell Signaling Technology (55 mentions)
Cyclin d1, by Cell Signaling Technology (53 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (49 mentions)
Ripa buffer, by Thermo Fisher Scientific (47 mentions)
1 510 protocols using vimentin
1
Comprehensive Immunohistochemical Profiling
2
Analyzing Molecular Mechanisms of Cancer
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The inhibitor of RNA polymerase II/III, α-amanitin, was purchased from MedChem Express (Monmouth Junction, NJ). The luciferin (in vivo grade) used for animal experiments was purchased from Promega (Madison, WI). For Western blot analysis, the primary antibodies against β-actin, c-Myc, BMI1, p27 Kip1, Cyclin D1, CDK2, CDK4, ERK1/2, p-ERK1/2(Thr202/Tyr204), ZEB1, β-Catenin, N-Cadherin, Vimentin, ATM, p-ATM, and Rad51 were all purchased from Cell Signaling Technology (Beverly, MA); HMMR#1 was purchased from OriGene (Rockville, MD); and HMMR#2 was purchased from GeneTex (Irvine, CA). For immunohistochemical (IHC) staining, the primary antibody HMMR#1 was purchased from Origene (Rockville, MD), ZEB1 was from Abcam (Cambridge, MA), and β-Catenin and Vimentin were purchased from Cell Signaling Technology (Beverly, MA).
3
Immunoblot and Immunofluorescence Assays
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The primary antibodies used in the immunoblotting against E-cadherin, Twist1, STAT6, STAT3, GAPDH and Actin were from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The N-cadherin, Vimentin, ZO-1, Snail1, Slug, ZEB1, MMP9, p-STAT6 (Tyr641), p-STAT3 (Tyr705), AKT, p-AKT, Erk1/2, p-Erk1/2, nanog and CD44 antibodies were from Cell signaling Technology (Beverly, MA, USA). Fibronectin antibody was from Millipore (Billerica, MA, USA). CD166 and histone H3 antibodies were from Abcam (Cambridge, UK). CD133 antibody was from MiltenyiBiotec (GmbH, Bergisch Gladbach, Germany). IL-13Rα1 and IL-13Rα2 antibody were from Sangon Biotech (Shanghai, China). The primary antibody used in the immunofluorescence against E-cadherin and Vimentin were from Cell signaling Technology (Beverly, MA, USA) and DAKO (Carpinteria, CA, USA), respectively. The pharmacological reagents used in this study are the following: recombinant human IL-13 (Peprotech, Rocky Hill, NJ, USA); JAK inhibitor 1 (JAKi1) and LY294002 (Calbiochem, San Diego, CA, USA).
4
Protein and Cellular Analyses of Tissues
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were lysed by incubation on ice for 20 min. Proteins were extracted with a protein extraction kit (Beyotime, Shanghai) and used for WB assays. Tissue samples were incubated with 10% neutral buffer and then embedded in formalin paraffin for IHC. Antibodies against alpha-SMA, FAP, Ki67, E-cadherin, Vimentin, LASP1, Anxa2, p-STAT3, STAT3, Nanog, CD9, CD68, Tsg101, Bcl-2 and Bax were purchased from Cell Signaling Technology (Danvers, MA, USA). An anti-beta-actin antibody was used as the control. For IF staining, cells were seeded on previously prepared coverslips and incubated for 24 hours. After being fixed with 4% paraformaldehyde, the cells were treated with 0.1% Triton-100 and then incubated with 5% goat serum. Then, the cells were stained with primary and secondary antibodies. Primary antibodies against Vimentin, alpha-SMA and FAP were purchased from Cell Signaling Technology (Danvers, MA, USA). The nuclei were stained with 4',6-diamidino-2-phenylindole (DAPI). Then, fluorescence was detected using a Nikon Eclipse 80i microscope.
5
Antibody Characterization for Biomedical Research
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Antibodies used for all the experiments were as follows: for immunoflourescence, β-hCG (ab9582, Abcam), Vimentin (5144 S, Cell signaling technology), E-cadherin (3195 S, Cell signaling technology), P-cadherin (sc-7893, Santa Cruz Biotechnology), BRCA1 (9010 S, Cell signaling technology) primary antibodies were followed by secondary antibodies conjugated with FITC (35552, Cell Signaling Technology). For immunoblotting, Vimentin (5144 S, Cell signaling technology), E-cadherin (3195 S, Cell signaling technology), β-actin (sc-47778, Santa Cruz Biotechnology). For immunoprecipitation: β-hCG (sc-271062, Santa Cruz Biotechnology), TGF beta Receptor II (ab78419, Abcam), normal IgG (sc-2025, Santa Cruz Biotechnology). ChIP: BRCA1 (A301-377 A, Bethyl laboratories). Immunohistochemical analysis: β-hCG (AM305-5M, Biogenex), BRCA1 (AR345-5 R, Biogenex).
6
Comprehensive Western Blot Analyses
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For western blot, samples were resolved by SDS-PAGE, transferred to nitrocellulose membranes, and blotted with various antibodies; anti-GAPDH (#2118), N-cadherin (#13116), E-cadherin (#3195), Vimentin (#5741), Slug (#9585), Snail-1 (#3879), ZEB1 (#3396), ZO-1 (#8193), phospho-Met (Tyr1349) (#3121), Ron (#4269), pβ-catenin (Thr41 or Ser45) (#9565), β-catenin (#8480), S6 (#2217), total Akt (#9272), phospho-c-Myc (Thr58/Ser62) (#9401), Ki-67 (#9027), phospho-AKT (Ser473) (#9271), phospho-EGFR (Tyr1068) (#11862), LRIG1 (#12752), c-Met (#8198) and antibodies were purchased from Cell Signaling Technology. LRIG1 (G-20) (#sc-50075) and EGFR (#sc-03) antibodies were purchased from Santa Cruz Biotechnology. c-Myc antibody (clone 9E11) (#MS-127-P0) was purchased from Neomarkers. Fibronectin (#GTX112794), CD44 (#GTX102111) and Twist (#GTX127310) were purchased from Genetex (CA, USA). Actin (#A5441) and Tubulin (#T5168) were purchased from Sigma (MO, USA). All antibodies used horseradish peroxidase-conjugated secondary antibodies (Biorad), followed by developing with SuperSignal West chemicals (Pierce). An AlphaInnotech imaging station with FluorChem software was used to capture images. All data are representative of more than three independent experiments.
7
Protein Expression Profiling in Cell Lysates
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were lysed with RIPA buffer (25 mM Tris HCl (pH 7.6), 1% sodium deoxycholate, 150 mM NaCl and 0.1% SDS) containing protease inhibitor cocktail (Roche, Basel, Switzerland). Total cell lysates with 60–80 µg protein were boiled for five minutes in SDS-PAGE reducing buffer, and loaded to SDS-PAGE gels. Separated proteins in the gel were transferred onto a nitrocellulose membrane. Primary antibodies used for immunoblotting were against androgen receptor (#3125, Cell Signaling Technology), vimentin (#5741, Cell Signaling Technology), fibronectin (15613-1-AP, Proteintech), Keratin 8/18 (#4546, Cell Signaling Technology), and GAPDH (#2118S, Cell Signaling Technology). Fluorescence-labeled secondary anti-mouse IgG antibody (LI-COR Biosciences, Lincoln, NE, USA) was used to detect the primary antibodies. The fluorescence intensity of the secondary antibody was assayed using an Odyssey Imaging System (LI-COR).
8
Proteomic Analysis of ST09 Treatment
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A total of 75,000 cells/mL were seeded and treated with ST09 (20, 40, 60, and 80 nM) for 48 h and the whole cell lysate was prepared as described [13 (link)]. Next, 30 µg of cell lysates was electrophoresed on 10 to 12% of SDS-PAGE (poly acrylamide gel electrophoresis) and were transferred to a polyvinylidene fluoride membrane (Millipore, Burlington, MA, United States). Blocking was performed using 5% skim milk in 1× PBS and then probed with primary antibodies: MMP2 from Biolegend, MMP1 from elabscience, Apaf, Bad, Bcl2, cytochrome c, Tubulin from Santa-Cruz Biotechnology, CA, and Caspase 9, Caspase 3, PARP, Vimentin, Bax, and GAPDH from Cell Signaling Technology, Beverly, MA, USA, followed by HRP-conjugated secondary anti-rabbit, anti-mouse antibodies (Cell Signaling Technology). The blots were developed using chemiluminescence reagent (Clarity Western ECL blotting substrate, Biorad) and the blot images were captured by the Chemidoc-XRS Biorad gel doc system. The protein band images were quantified using GelQuant.Net, BiochemLab solutions.
9
Western Blot Analysis of Cell Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were lysed in lysis buffer, and an equal amount of protein was loaded into each well of a sodium dodecyl sulfate (SDS)-PAGE (12%) gel. Proteins were resolved by an applied potential and then blotted to polyvinylidene difluoride (PVDF) membranes (Merck Millipore, Burlington, MA). Blots were rinsed, blocked for 1 h, and probed at 4 °C with primary antibodies against: (i) PTPRA (1:1000; Proteintech, Rosemont, IL), (ii) RASSF8(1:1000; Proteintech), (iii) N-cadherin (1:1000; Cell Signaling Technologies, Danvers, MA), (iv) E-cadherin (1:1000; Cell Signaling Technologies), (v) ALIX (1:1000; Proteintech), (vi) CD63 (1:1000; Abcam, Cambridge, UK), (vii) heat shock protein 70 (HSP70, 1:2000; Proteintech), (viii) tumor susceptibility gene 101 TSG101 (1:1000; Proteintech), (ix) GAPDH (1:5000; Proteintech), and (x) vimentin (1:1000; Cell Signaling Technologies).
The following day, blots were rinsed and probed for 1 h at room temperature (RT) with horseradish peroxidase (HRP)-tagged secondary antibody. Following additional rinse step, blots were developed with a Pierce™ ECL (enhanced chemiluminescence) Western Blotting Substrate (Thermo Fisher Scientific) and detected bands were quantified using Quantity One® 1-D Analysis Software (Bio-Rad, USA).
The following day, blots were rinsed and probed for 1 h at room temperature (RT) with horseradish peroxidase (HRP)-tagged secondary antibody. Following additional rinse step, blots were developed with a Pierce™ ECL (enhanced chemiluminescence) Western Blotting Substrate (Thermo Fisher Scientific) and detected bands were quantified using Quantity One® 1-D Analysis Software (Bio-Rad, USA).
10
Analyzing STIM1 and HIF-1α expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
RNA extraction and q-PCR were performed as described previously25 . Expression of STIM1 and HIF-1α was detected using a TaqMan q-PCR assay system (Applied Biosystems, Foster City, CA, USA). Western blot was performed as previous described25 . The antibodies used in this study were: STIM1 (Cell Signaling Technology, 5668), HIF-1α (Abcam, 113642), E-cadherin (Cell Signaling Technology, 3195), and vimentin (Cell Signaling Technology, 49636).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!