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TUBB3 is a gene that provides instructions for making the beta-III tubulin protein, which is a component of microtubules. Microtubules are cytoskeletal structures that play a crucial role in various cellular processes, such as cell division, intracellular transport, and the maintenance of cell shape and structure. The TUBB3 gene is commonly used as a target for research and diagnostic applications in the life sciences industry.

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Lab products found in correlation

Taqman primer, by Thermo Fisher Scientific (2 mentions) Pou5f1, by Thermo Fisher Scientific (2 mentions) Vegfa, by Santa Cruz Biotechnology (1 mentions) Hrp conjugated goat anti rabbit igg antibody, by GE Healthcare (1 mentions) Sc 372, by Santa Cruz Biotechnology (1 mentions) Evos fl auto fluorescent microscope, by Thermo Fisher Scientific (1 mentions) Species specific fluorescent secondary antibodies, by Thermo Fisher Scientific (1 mentions) Tubb3, by Fortrea (1 mentions) Nanodrop spectrophotometer, by Thermo Fisher Scientific (1 mentions) Eif2ak2, by Thermo Fisher Scientific (1 mentions) Taqman reverse transcription reagent, by Thermo Fisher Scientific (1 mentions) High pure rna isolation kit, by Roche (1 mentions)

5 protocols using tubb3

1

Western Blot Analysis of Lung Proteins

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Proteins were extracted from snap‐frozen left lobes of the lung and analyzed according to previously described Western blot procedures 5. The antibodies were used at the following concentrations for 1 hour at RT: vascular endothelial growth factor A (VEGFA, 1:200, Santa Cruz), β‐tubulin (1:1,000, TUBB3, Thermo Scientific), and Pro‐surfactant protein C (SFTPC, 1:2,000, Millipore, Billerica, MA). Secondary HRP‐conjugated goat anti‐rabbit IgG antibody (1:15,000, GE Healthcare, Buckinghamshire, U.K., http://www.gehealthcare.com/) was used for 1 hour at RT.
Firsova A.B., Bird A.D., Abebe D., Ng J., Mollard R, & Cole T.J. (2017). Fresh Noncultured Endothelial Progenitor Cells Improve Neonatal Lung Hyperoxia‐Induced Alveolar Injury. Stem Cells Translational Medicine, 6(12), 2094-2105.
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2

Fluorescent Immunostaining of RGCs and AAV-293 Cells

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Primary RGCs and AAV-293 cells were fixed in 4% paraformaldehyde (PFA) for 30 minutes, rinsed in PBS (3 × 10 minutes) and blocked with 5% normal donkey serum with 0.15% Tween-20 in PBS at pH 7.4. Cells were then incubated with either both beta III Tubulin antibody (Tubb3, 1:250, Covance, Denver, PA) and p65 antibody (1:400; sc-372, Santa Cruz Biotechnology, Dallas, TX) or with Tubb3 alone, followed by species-specific secondary fluorescent antibodies (Thermo Fisher Scientific, Grand Island, NY). Negative controls were incubated with secondary antibodies only. The cells were imaged using an EVOS FL Auto fluorescent microscope with onstage incubation capabilities (Thermo Fisher Scientific, Grand Island, NY).
Dvoriantchikova G., Pappas S., Luo X., Ribeiro M., Danek D., Pelaez D., Park K.K, & Ivanov D. (2016). Virally delivered, constitutively active NFκB improves survival of injured retinal ganglion cells. The European journal of neuroscience, 44(11), 2935-2943.
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3

Stem Cell Characterization by qPCR

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RNA was isolated via QIAcube (serial number: 50946) and RNeasy Mini Kit protocol “Purification of total RNA from 5 × 106 − 1 × 107 animal cells (with QIAshreddder homogenization)” (Firmware Version FIW-50–001-J_FW_MB.hex and program version FIW-50–002-G_PLC_MB.prs or higher) following manufacturer’s instructions. cDNA was synthesized using the High-Capacity cDNA Reverse Transcription Kit (Applied Biosysystems, Cat. # 4368814) according to manufacturer’s instructions. Quantitative PCR with the Taqman™ primers POU5F1 (Thermofisher, Cat. # 4448892), PAX6 (Thermofisher, Cat. # 4448892), NES (Thermofisher, Cat. # 4448892), TUBB3 (Thermofisher, Cat. # 4448892), and OTX2 (Thermofisher, Cat. # 4448892) was carried out by a liquid handling robotic system and run via Quantstudio 12K Flex (serial number: 285880333).
Kuang Y.L., Munoz A., Nalula G., Santostefano K.E., Sanghez V., Sanchez G., Terada N., Mattis A.N., Iacovino M., Iribarren C., Krauss R.M, & Medina M.W. (2019). Evaluation of commonly used ectoderm markers in iPSC trilineage differentiation. Stem cell research, 37, 101434.
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4

Stem Cell Characterization by qPCR

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RNA was isolated via QIAcube (serial number: 50946) and RNeasy Mini Kit protocol “Purification of total RNA from 5 × 106 − 1 × 107 animal cells (with QIAshreddder homogenization)” (Firmware Version FIW-50–001-J_FW_MB.hex and program version FIW-50–002-G_PLC_MB.prs or higher) following manufacturer’s instructions. cDNA was synthesized using the High-Capacity cDNA Reverse Transcription Kit (Applied Biosysystems, Cat. # 4368814) according to manufacturer’s instructions. Quantitative PCR with the Taqman™ primers POU5F1 (Thermofisher, Cat. # 4448892), PAX6 (Thermofisher, Cat. # 4448892), NES (Thermofisher, Cat. # 4448892), TUBB3 (Thermofisher, Cat. # 4448892), and OTX2 (Thermofisher, Cat. # 4448892) was carried out by a liquid handling robotic system and run via Quantstudio 12K Flex (serial number: 285880333).
Kuang Y.L., Munoz A., Nalula G., Santostefano K.E., Sanghez V., Sanchez G., Terada N., Mattis A.N., Iacovino M., Iribarren C., Krauss R.M, & Medina M.W. (2019). Evaluation of commonly used ectoderm markers in iPSC trilineage differentiation. Stem cell research, 37, 101434.
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5

Quantitative mRNA Expression Analysis

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Total RNA purification was performed using High Pure RNA Isolation Kit (Roche Life Science) with an on‐column enzymatic DNA degradation, according to the manufacturer's instructions. RNA concentration was determined on a Nanodrop spectrophotometer and cDNA created using equal amounts of RNA and the TaqMan® Reverse Transcription Reagents with random primers according to manufacturer's instructions (Applied Biosystems). Quantitative PCR was performed using the iTaq™ SYBR® Green Mastermix with ROX (BioRad) with the following probes: NADH (NDUFC2, Hs01072843_m1), tau (MAPT, Hs00902194_m1), PKR (EIF2AK2, Hs00169345_m1), β‐actin (ActB, Mm00607939_s1), IFN‐β (Ifnb1, Mm00439552_s1), TNF‐α (Tnf, Mm00443260_g1), tau (MAPT, Mm00521988_m1), neuron‐specific class III beta‐tubulin (Tubb3, Mm00727586_s1) and PKR (EIF2AK2, Mm01235643_m1, all Thermo Fisher Scientific Inc.).
Reimer L., Betzer C., Kofoed R.H., Volbracht C., Fog K., Kurhade C., Nilsson E., Överby A.K, & Jensen P.H. (2020). PKR kinase directly regulates tau expression and Alzheimer's disease‐related tau phosphorylation. Brain Pathology, 31(1), 103-119.
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