The purified polyclonal antibody against SNPH residues 225–428 was described previously (Kang et al., 2008 (link)). Sources of other antibodies or reagents are as follows: anti–βIII-tubulin, anti-MAP2, anti–Tau-1, anti-dynein IC74, and anti–GAP-43 were from EMD Millipore; anti–cytochrome c was from BD; anti-TOM20 was from Santa Cruz Biotechnology, Inc.; anti-Miro1/2 (HPA010687) and anti-Trak2 were from Sigma-Aldrich; ECL-HRP–linked secondary antibodies were from GE Healthcare; and Alexa Fluor 546– or Alexa Fluor 488–conjugated secondary antibodies were from Invitrogen.
Anti βiii tubulin
Manufactured by Merck Group
Sourced in United Kingdom, Japan
Anti-βIII-tubulin is a monoclonal antibody used in research applications to detect and quantify the expression of the βIII-tubulin protein, which is a component of the cytoskeleton. This antibody can be used in techniques such as Western blotting, immunocytochemistry, and immunohistochemistry to study the expression and localization of βIII-tubulin in various cell types and tissues.
Automatically generated - may contain errors
Lab products found in correlation
Bovine serum albumin (bsa), by Merck Group (6 mentions)
Anti gfap, by Agilent Technologies (5 mentions)
Alexa fluor 488, by Thermo Fisher Scientific (5 mentions)
Anti map2, by Merck Group (3 mentions)
Anti gaba, by Merck Group (3 mentions)
Anti sox2, by Merck Group (3 mentions)
Anti β actin, by Merck Group (2 mentions)
Triton x 100, by Merck Group (2 mentions)
Ecl western blotting detection reagent, by Merck Group (2 mentions)
Anti gad67, by Merck Group (2 mentions)
Las 1000plus, by Fujifilm (2 mentions)
Quantity one 1 d analysis software, by Bio-Rad (2 mentions)
Alexa fluor 594 or 488 secondary antibodies, by Thermo Fisher Scientific (2 mentions)
Dfc365fx, by Leica (2 mentions)
Anti nanog, by Abcam (2 mentions)
Anti gfap, by Merck Group (2 mentions)
Lsm 510 meta microscope, by Zeiss (2 mentions)
Anti nestin, by Abcam (2 mentions)
Anti nestin, by Merck Group (2 mentions)
Anti βii tubulin, by Fortrea (2 mentions)
43 protocols using anti βiii tubulin
1
Immunofluorescence Staining Antibody Protocol
2
Pluripotent Stem Cell Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
AP staining was performed using an ES-AP detection kit (EMD Millipore) according to manufacturer’s recommendations. SA-β-gal activity was detected using SA-β-gal staining kit (Cell Signaling Technology) according to manufacturer’s protocol. For immunocytochemical analysis, we used anti-SSEA4, TRA-1-60, TRA-1-81 (mouse, 1:100; EMD Millipore), anti–β-III-tubulin (mouse, 1:400; EMD Millipore), rabbit anti–LC-3B (1:200; Cell Signaling Technology), rabbit anti-ASM (1:1,000, Abcam), mouse anti-LAMP1 (1:100; Abcam), and mouse anti-LBPA (1:500; Echelon).
3
Immunofluorescence Analysis of Endothelial Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Differentiated ES cells were fixed with phosphate-buffered saline 4% paraformaldehyde (PBS-PFA) (Santa Cruz), treated for 1 h with PBS 0.1% Triton X100 (Merck Life Science) containing 5% donkey serum (Dako North America Inc) and 5% BSA (Merck Life Science) and incubated with primary antibodies for 3 h at 37 °C in a moist chamber. The following primary antibodies were used: anti-βIII-tubulin 1:1000 diluted in PBS (Merck Life Science) and anti-VE-cadherin 1: 100 diluted in PBS (R&D Systems). After rinsing three times with PBS, cells were incubated with 555 AlexaFluor donkey anti-goat and 488 AlexaFluor donkey anti-mouse (Invitrogen, Thermofisher Scientific) at 5 µg/ml for 1 h. After rinsing three times with PBS, nuclei were stained with DAPI and slides were sealed with aqueous mounting solution (Dako North America Inc). Images were captured by using a Leica TCS SPE confocal laser-scanning microscope, analyzed with Leica Confocal Software (LCS; Leica Microsystems). To quantify VE-cadherin+ endothelial cells, sequential images, covering the entire surface of a chamber slide well (0.7 cm2), were acquired with a Leica AF6000LX workstation. The area occupied by endothelial cells was measured with ImageJ software by using the Angiogenesis Analyzer for ImageJ [25 ].
4
Immunohistochemical Characterization of Glial and Neural Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Glial fibrillary acidic protein (GFAP) was detected by chicken polyclonal anti-GFAP (cat. no. AB5541; Merck KGaA). Other antibodies were as follows: Rabbit polyclonal anti-light chain (LC)3B (cat. no. 2775), rabbit monoclonal anti-Atg7 (D12B11; cat. no. 8558), anti-cyclin dependent kinase (CDK)2 (78B2; cat. no. 2546; Cell Signaling Technology, Inc.) and anti-p62 (cat. no. P0067), mouse monoclonal anti-nestin (cat. no. N5413) (both Sigma-Aldrich; Merck KGaA), anti-βIII-tubulin (cat. no. AB15708; Merck KGaA), anti-O4 (cat.no. O7139; Sigma-Aldrich; Merck KGaA), anti-β-actin (cat. no. sc-47778; Santa Cruz Biotechnology, Inc.), anti-tubulin βIII (cat. no. MAB1637; Merck KGaA), anti-doublecortin (DCX; cat. no. ab2253) and anti-caspase-3 antibody (cat. no. 9662; Cell Signaling Technology, Inc.). Curcumin (cat. no. C1386-5G) and monodansylcadaverine (MDC) were purchased from Sigma-Aldrich; Merck KGaA (cat. no. D4008-100MG). 3-Methyladenine (3MA, an inhibitor of phosphatidylinositol 3-kinases and autophagosome formation; cat. no. s2767) was purchased from Selleck Chemicals (Shanghai, China).
5
Immunofluorescence Analysis of Cilia
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cells were washed with Dulbecco's Phosphate-Buffered Saline (DPBS, Welgene, Korea) and fixed in 4% formaldehyde (Electron Microscopy Sciences, Hatfield, PA, USA) for 10 min at room temperature. After washing with DPBS, the cells were blocked and permeabilized with 3% BSA and 0.3% TritonX-100 (Sigma-Aldrich) in DPBS for 1 h at room temperature.
The cells were stained with anti-Arl13b (1:100~250) in 1% BSA in DPBS and washed with 0.1% BSA in DPBS, followed by incubation in the proper secondary antibody. If needed, anti-βIII tubulin (Merck Millipore, AB9354) was used for staining. Hoechst33342 was used to visualize the nuclei. Cell images were taken with a confocal microscope or a FLoid cell imaging station (Thermo Fisher Scientific) and the ciliated cells were counted by two persons blinded to the treatments. We used Arl13B as a cilia marker for the entire study. To obtain the percentage of ciliated cells, the number of Arl13B+ cells was counted as ciliated cells and divided by the total number of cells.
The cells were stained with anti-Arl13b (1:100~250) in 1% BSA in DPBS and washed with 0.1% BSA in DPBS, followed by incubation in the proper secondary antibody. If needed, anti-βIII tubulin (Merck Millipore, AB9354) was used for staining. Hoechst33342 was used to visualize the nuclei. Cell images were taken with a confocal microscope or a FLoid cell imaging station (Thermo Fisher Scientific) and the ciliated cells were counted by two persons blinded to the treatments. We used Arl13B as a cilia marker for the entire study. To obtain the percentage of ciliated cells, the number of Arl13B+ cells was counted as ciliated cells and divided by the total number of cells.
6
Immunofluorescence Analysis of Neuronal Markers
7
Western Blot Analysis of Hippocampal Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mouse hippocampus were prepared from P30 control and CCK-Cre;Erbb4F/F homogenized in lysis buffer containing 50 mM Tris pH 7.5, 150 mM NaCl, 5 mM EDTA, 1% Triton X-100, 0.5 mM prevanadate with Protease Inhibitor Cocktail (Complete, Mini, Roche). Samples were denatured and run on 10% SDS–PAGE gels. Gels were electrophoretically transferred onto PVDF membranes (Whatman GmbH). Membranes were blocked with 5% BSA (Sigma) in TBS (20 mM Tris-HCl, pH 7.5, 150 mM NaCl) for 1 h and probed with primary antibodies: anti-β III-tubulin (1:2000; Sigma #T8660), anti-GAD65 (1:1000; Millipore #MAB351R) and anti-GAD67 (1:1000; Chemicon #MAB5406), in 1% BSA in TBS + 0.1% Tween20. Subsequently, they were treated with horseradish-peroxidase-conjugated secondary antibodies and ECL western blotting detection reagents (Immobilon, Millipore) Signals were acquired as 16 bit images with a luminescent image analyzer (LAS-1000PLUS; Fujifilm) and quantified with Quantity One 1D Analysis Software (Bio-Rad Laboratories).
8
Immunocytochemical Analysis of α-Synuclein
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For immunocytochemical staining, SH-SY5Y cells or primary hipocampal neurons were plated onto cover slips coated with poly-L -lysine. Cells were rinsed with PBS, fixed with 4% paraformaldehyde and permeabilized with Tris-HCl buffer (pH 7.4) containing 0.25% Triton for 20 min. Nonspecific epitopes were blocked with 1% BSA. Then, incubations with primary antibodies were carried out in buffer containing 0.1% Triton at 4°C for 16 h. The following antibodies were used: anti-α-syn (1 : 1000; BD Transduction Laboratories), anti-phospho-Ser129 α-syn (1 : 200; ab51253 from Abcam plc, Cambridge, UK) and anti-β-III tubulin (1 : 5000; Sigma Aldrich). Phospho-Ser129 α-syn was detected using biotinylated goat-anti-rabbit (1 : 200; Vector Laboratories, Burlingame, CA, USA) and streptavidin-conjugated Dylight fluorophore 488 (1 : 400; Vector Laboratories). α-Syn and β-III Tubulin were detected directly with Dylight 594 (1 : 400; Vector Laboratories) secondary antibody. In some experiments, nuclear staining was achieved by incubation with 4',6-diamidino-2-phenylindole (Dapi) (Biotium Inc., Hayward, CA, USA). Images were acquired with a Zeiss LSM 710 NLO confocal microscope (Jena, Germany).
9
Immunocytochemistry Analysis of Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For immunocytochemistry, cells were fixed with 4% paraformaldehyde for 20 minutes at RT, washed PBS, and blocked with 10% bovine serum and 0.1% Triton X-100. Primary antibodies include anti β-III TUBULIN (T2200, Sigma Aldrich), anti Mitochondria (NB600-556, Novus Biologicals). Secondary antibodies used were conjugated with Alexa 488 or Alexa 555 (Invitrogen) and nuclei were counterstained with 1 μg/ml Hoechst 33342 (Invitrogen). Coverslips were mounted using PBS/Glycerol (1:1), visualized using a confocal microscope Fluoview FV1000 (Olympus) and acquired with the software FV10-ASW Version 2.0.
10
Immunofluorescence Analysis of ESCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For immunofluorescence analysis, ESCs were fixed, permeabilized, and incubated with primary antibodies and an appropriate secondary antibody (13 (link)). Nuclei were counterstained with 4',6-diamidino-2-phenylindole (1:5000; Calbiochem). Sectioned SFEBs were obtained and stained as previously described (15 (link)). The following primary antibodies were used: anti-Yap (1:300; NB110-58358; Novus Biological) and anti-βIII tubulin (1:400; Sigma). Alexa Fluor 594 or 488 secondary antibodies were used (1:400; Thermo Fisher Scientific). Cells were visualized using an inverted microscope (Leica Microsystems), and the images were captured with a digital camera (DFC365 FX; Leica Microsystems) using LAS-AF software (Leica Microsystems).
Confocal images were acquired with LSM510META microscope (Carl Zeiss GmbH) using LSM510 software (Zeiss). After acquisition, the images were color corrected using the brightness, contrast, and color-balance commands applied to every pixel in each image.
Confocal images were acquired with LSM510META microscope (Carl Zeiss GmbH) using LSM510 software (Zeiss). After acquisition, the images were color corrected using the brightness, contrast, and color-balance commands applied to every pixel in each image.
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!