Polycaprolactone diol (PCL, MW∼2000) and N-hydroxy succinimide (NHS) were purchased from Sigma-Aldrich (Beijing, China). Branched polyethyleneimine (PEI, MW 1800) and N,N′-dicyclohexylcarbodiimide (DCC) were purchased from Alfa Aesar (Shanghai, China). Dithiodiglycolic acid, 4-dimethylaminopyridine (DMAP) was purchased from TCI (Shanghai, China). Hyaluronic acid (HA) was purchased from Macklin (Shanghai, China). N, N-dimethylformamide (DMF) and dimethyl sulfoxide (DMSO) were purchased from Aladdin (Shanghai, China). Lonidamine (LND) was purchased from Meilunbio (Dalian, China). BPTES was purchased from Famo biotechnology (Shanghai, China). Primary antibodies including rabbit anti-Bax, rabbit anti-Bcl-2, rabbit anti-PARP, rabbit anti-cytochrome c, rabbit anti-P21, rabbit anti-Cyclin D1, rabbit anti-Cyclin E2 and rabbit anti-Tubulin were obtained from Proteintech (Wuhan, China). Rabbit anti-Ki67 was purchased from Cell Signaling Technology, Inc. Secondary antibody of horseradish peroxidase-conjugated goat IgG was purchased from Beyotime (Jiangsu, China).
Rabbit anti tubulin
Manufactured by Proteintech
Sourced in United States
Rabbit anti-Tubulin is an antibody that specifically recognizes tubulin, a structural protein found in the cytoskeleton of eukaryotic cells. This antibody can be used for the detection and analysis of tubulin in various experimental techniques, such as immunoblotting, immunohistochemistry, and immunocytochemistry.
Automatically generated - may contain errors
Lab products found in correlation
N hydroxysuccinimide, by Merck Group (1 mentions)
N n dicyclohexylcarbodiimide, by Thermo Fisher Scientific (1 mentions)
Hyaluronic acid, by Maclin Biochemical Technology (1 mentions)
Rabbit anti bax, by Proteintech (1 mentions)
Rabbit anti parp, by Proteintech (1 mentions)
Rabbit anti p21, by Proteintech (1 mentions)
Rabbit anti cyclin d1, by Proteintech (1 mentions)
Rabbit anti ki67, by Cell Signaling Technology (1 mentions)
Polycaprolactone diol, by Merck Group (1 mentions)
Rabbit anti bcl 2, by Proteintech (1 mentions)
Mouse anti gapdh, by Proteintech (1 mentions)
Enhanced chemiluminescence detection system, by Thermo Fisher Scientific (1 mentions)
Bradford method, by Bio-Rad (1 mentions)
Pvdf membrane, by Bio-Rad (1 mentions)
Alexa 488, by Thermo Fisher Scientific (1 mentions)
Alexa 647, by Thermo Fisher Scientific (1 mentions)
Rabbit anti gfp, by Santa Cruz Biotechnology (1 mentions)
Mouse anti ha clone 16b12, by Fortrea (1 mentions)
Rabbit anti tgf β1, by Proteintech (1 mentions)
Rabbit anti wnt5a, by Proteintech (1 mentions)
6 protocols using rabbit anti tubulin
1
Synthesis and Characterization of Polycaprolactone-Based Nanoparticles
2
Quantifying NFIL3 and NFKBIA in Breast Cancer
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Total proteins were extracted from breast cancer cells using RIPA lysis buffer and subsequently quantified using the Bradford method (Bio–Rad Laboratories, Hercules, CA, USA). Twenty micrograms of protein were loaded and separated by polyacrylamide gel electrophoresis and then transferred to PVDF membranes (Bio–Rad). After blocking with 5% BSA, membranes were incubated with rabbit anti-NFIL3 (1:1000, Cat.#11,773–1-AP, Proteintech, Rosemont, IL, USA), rabbit anti-NFKBIA (1:1000, Cat.#10,268–1-AP, Proteintech), mouse anti-GAPDH (1:5000, Cat.#600,04–1-Ig, Proteintech) or rabbit anti-tubulin (1:5000, Cat.#66,031–1-Ig, Proteintech) at 4 °C overnight. Then, membranes were incubated with peroxidase-conjugated goat anti-rabbit or goat anti-mouse secondary antibody for 1 h after washing. Finally, proteins were visualized through an enhanced chemiluminescence detection system (Thermo Scientific, Rockford, IL, USA). The relative gray value was measured using Image J software.
3
Importin Antibody Validation Protocol
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The antibody against importin 7 was raised in rabbits against a peptide from the C-terminal region of the protein (CLADQRRAAHESKMIEKHG) and affinity-purified. The antibody against importin β was raised in rabbits against the full-length protein and affinity purified. Antibodies against Nup358 were described previously [30] (link). Rabbit-anti-tubulin was from Proteintech (11224-1-AP). For the detection of tagged proteins, mouse-anti-myc (clone 9E10, Serotec), mouse-anti-HA (clone 16B12, Covance) and rabbit-anti-GFP (Santa Cruz) were used. For immunofluorescence, secondary antibodies from donkey, coupled to Alexa 488, Alexa 594 or Alexa 647 were used (1∶2000; Molecular Probes). For immunoblotting, HRP-coupled donkey anti-rabbit IgG (Dianova) was used as secondary antibody.
4
Western Blotting Analysis of Cell Signaling Proteins
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The cultured cells were collected and lysed in RIPA buffer (Beyotime) with protease inhibitors (Beyotime) after rinsing twice with PBS (precooled at 4°C). Protein was quantified with a BCA protein concentration determination kit (Beyotime), and then separated by SDS- polyacrylamide gel electrophoresis and electroblotted onto polyvinylidene fluoride membranes. After blocking in 5% nonfat milk for 2 h at 25°C, the membranes were probed with primary antibodies followed by horseradish peroxidase-conjugated secondary antibodies (Proteintech Group, Inc.). Primary antibodies used were rabbit anti-VEGF protein (1 : 1000, Proteintech Group, Inc.), rabbit anti-Wnt5a (1 : 1000, Proteintech Group, Inc.), rabbit anti-TGF-β1 (1 : 1000, Proteintech Group, Inc.) and rabbit anti-Tubulin (l : 1000, Proteintech Group, Inc.). We used BeyoECL Star (Beyotime) reagent to detect protein.
5
Protein Analysis of HUVEC and Tumor Tissue
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HUVECs were washed twice with ice-cold PBS and lysed with cell lysis buffer (Beyotime Biotechnology) containing an EDTA-free protease inhibitor (Roche). Tumor tissue was incubated with ice-cold cell lysis buffer containing protease inhibitor and disrupted with Tissuelyser-24. Protein concentration was quantified using the Pierce™ BCA Protein Assay Kit (Thermo Fisher Scientific, Waltham, MA, USA). Western blotting was performed as previously described (23 (link)). Blots were incubated with Pierce ™ ECL Western Blotting Substrate (Thermo Fisher), and detected by AI 680 (General Electric Company, Boston, MA) Antibodies used for western blotting included rabbit anti-phospho-STAT6 (Tyr-641, 1:1000, Cell Signaling Technology); rabbit anti-STAT6 (1:1000, Abcam); rabbit anti-NRP1 (1:1000, Affinity); rabbit anti-NRP1 (1:1000, Abcam); and rabbit anti-Tubulin (1:5000, Proteintech). The secondary antibody was horseradish peroxidase (HRP)-conjugated goat anti-rabbit immunoglobulin G (IgG) (1:8000, Proteintech).
6
Western Blot Analysis of SH3GL2 Expression
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Cell, tumor, and spleen tissues were lysed in RIPA buffer with PMSF. Equal amounts of protein were separated by the sodium dodecyl sulfate/polyacrylamide electrophoresis gel and then the protein was transferred to the nitrocellulose polyvinylidene fluoride membrane. Different primary antibodies were used: rabbit anti-SH3GL2 (1:1000, cat. no12435-1-AP; Proteintech, Chicago, IL, USA) and rabbit anti-TUBULIN (1:1,000; cat. no. sc-5274; Santa Cruz Biotechnology, Dallas, TX, USA). The proteins were then incubated with horseradish peroxidase–conjugated anti-rabbit IgG (1:10000, cat. no. 7074s; Cell Signaling Technology, Inc., Danvers, MA, USA) for one hour at 37°C, and the membranes were then visualized by an enhanced chemiluminescence detection system.
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