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Anti β tubulin 4

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Anti-β-tubulin IV is a laboratory reagent used in the detection and analysis of the β-tubulin IV protein, which is a component of the cytoskeleton in cells. It is commonly used in various cell biology and biochemistry applications, such as immunocytochemistry, western blotting, and flow cytometry.

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Lab products found in correlation

Anti muc5ac, by Thermo Fisher Scientific (3 mentions) Vectashield antifade mounting medium, by Vector Laboratories (3 mentions) Alexa fluor 488, by Thermo Fisher Scientific (2 mentions) Axioobserver z1, by Leica (2 mentions) Rnaseout, by Thermo Fisher Scientific (2 mentions) Facsaria sorter, by BD (1 mentions) First strand synthesis kit, by Thermo Fisher Scientific (1 mentions) Rnaseout, by Merck Group (1 mentions) Spe confocal microscope, by Leica (1 mentions) Alexa fluor 488 or 594, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Anti-tubulin (542 citations) β-tubulin (395 citations) Anti-β-tubulin (260 citations) β-tubulin-IV (2 citations)

4 protocols using anti β tubulin 4

1

Immunofluorescence Staining of ALI-Cultured NHBE Cells

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Immunofluorescence staining on ALI-cultured NHBE cells was performed as described previously with some modifications83 (link). First, cells were fixed using 4% paraformaldehyde. Then, cells were blocked with PBS supplemented with 5% BSA and 0.2% Triton X-100 for 1 hour at room temperature. Primary antibody incubation was performed overnight at 4C in PBS supplemented with 1% BSA and 0.2% Triton X-100 using anti-β-tubulin IV (Sigma) or anti-MUC5AC (Thermo) at 1:100 dilution. Secondary antibodies conjugated with Alexa-fluor 488 (Life Technologies) were used at 1: 100 dilution. 4′-6-Diamidino-2-phenylindole, dihydrochloride was used to label the nuclear DNA and samples were mounted with Vectashield antifade mounting medium (Vector Labs, Burlingame, Calif). Confocal images were taken using Zeiss AxioObserver Z1 or Leica STP8000 and processed using ImageJ.
Park H.R., O’Sullivan M., Vallarino J., Shumyatcher M., Himes B.E., Park J.A., Christiani D.C., Allen J, & Lu Q. (2019). Transcriptomic response of primary human airway epithelial cells to flavoring chemicals in electronic cigarettes. Scientific Reports, 9, 1400.
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2

Intracellular Sorting and RNA Extraction

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Intracellular sorting followed by RNA extraction was adapted from a previously described protocol 30 (link). NHBE cells in ALI culture (day 21) were trypsinized and fixed with 4% paraformaldehyde in PBS with RNASEout (1:100, Life Technologies) for 30 minutes at room temperature. Following 2 washes with wash buffer (PBS supplemented with 0.5% BSA, 0.05% Tween-20 and 1:100 RNASEout), cells were incubated with either anti-β-tubulin IV (Sigma) or anti-MUC5AC (Thermo Scientific) antibodies for 1 hour at 4°C in PBS with 1% BSA, 0.05% Tween-20 and RNASEout (1:25). Following 3 washes with wash buffer, cells were incubated with secondary antibody (1:100), washed again for another 3 washes and sorted by FACS (BD FACS-Aria Sorter). RNA was then extracted using the PFFE Kit (Qiagen) and transcribed using random hexamers in the First-strand Synthesis Kit (Life Technologies). Quantitative PCR was performed for GSDMB using primers above and with the following additional primers (FOXJ1 and MUC5AC) to determine whether efficient separation of ciliated or goblet cells was achieved.
Panganiban R.A., Sun M., Dahlin A., Park H.R., Kan M., Himes B.E., Mitchel J.A., Iribarren C., Jorgenson E., Randell S.H., Israel E., Tantisira K., Shore S., Park J.A., Weiss S.T., Wu A.C, & Lu Q. (2018). A functional splicing variant associated with decreased asthma risk abolishes the ability of gasdermin B (GSMDB) to induce epithelial cell pyroptosis. The Journal of allergy and clinical immunology, 142(5), 1469-1478.e2.
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3

Immunofluorescence Staining of NHBE Cells

Cited 2 times
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Immunofluorescence staining on ALI-cultured NHBE cells was performed as described previously with some modifications29 (link),62 (link). First, cells were fixed using 4% paraformaldehyde. Then, cells were blocked with PBS supplemented with 5% BSA and 0.2% Triton X-100 for 1 h at room temperature. Primary antibody incubation was performed overnight at 4 °C in PBS supplemented with 1% BSA and 0.2%Triton X-100 using anti-β-tubulin IV (Sigma) or anti-MUC5AC (Thermo) at 1:100 dilution. Secondary antibodies conjugated with Alexa-fluor 488 (Life Technologies) were used at 1:100 dilution. 4′-6-Diamidino-2-phenylindole, dihydrochloride was used to label the nuclear DNA and samples were mounted with Vectashield antifade mounting medium (Vector Labs, Burlingame, Calif). Confocal images were taken using Zeiss AxioObserver Z1 or Leica STP8000 and processed using ImageJ.
Park H.R., Vallarino J., O’Sullivan M., Wirth C., Panganiban R.A., Webb G., Shumyatcher M., Himes B.E., Park J.A., Christiani D.C., Allen J, & Lu Q. (2021). Electronic cigarette smoke reduces ribosomal protein gene expression to impair protein synthesis in primary human airway epithelial cells. Scientific Reports, 11, 17517.
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4

Immunofluorescence Staining of NHBE Cells and Lung Tissue

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Immunofluorescence staining on ALI-cultured NHBE cells and paraffin-embedded lung tissue sections was performed as described previously with some modifications.31 (link) For ALI-cultured HNBE cells, fixation was performed in situ on inserts using 4% paraformaldehyde. For lung tissue sections, which were received from the Marsico Lung Institute/Cystic Fibrosis Center at the University of North Carolina, Chapel Hill, deparaffinization was performed by processing through a series of ClearRite and ethanol solutions. Antigen retrieval was performed using 10mM citrate buffer, pH 6.0. Both ALI-cultured NHBE cells and lung tissue sections were blocked with PBS supplemented with 5% normal donkey serum and 0.2% Triton X-100 for 1 hour at room temperature. Primary antibody incubation was performed overnight at 4°C in PBS supplemented with 1% BSA and 0.2% Triton X-100 using anti-GSDMB (Abgent) and anti-β-tubulin IV (Sigma) at 1:250 dilution. Secondary antibodies conjugated with Alexa-fluor 488 or 594 (Life Technologies) were used at 1:100 dilution. DAPI was used to label the nuclear DNA and samples were mounted with Vectashield anti-fade mounting medium (Vector Labs). Confocal images were taken using Leica SPE Confocal Microscope and the images were processed using ImageJ.
Panganiban R.A., Sun M., Dahlin A., Park H.R., Kan M., Himes B.E., Mitchel J.A., Iribarren C., Jorgenson E., Randell S.H., Israel E., Tantisira K., Shore S., Park J.A., Weiss S.T., Wu A.C, & Lu Q. (2018). A functional splicing variant associated with decreased asthma risk abolishes the ability of gasdermin B (GSMDB) to induce epithelial cell pyroptosis. The Journal of allergy and clinical immunology, 142(5), 1469-1478.e2.
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