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ITGB4 is a protein that functions as a receptor for laminin and plays a role in cell-matrix adhesion. It is commonly used as a marker for specific cell types and can be detected using various laboratory techniques.

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Lab products found in correlation

Biotinylated horse anti rabbit antibody, by Vector Laboratories (2 mentions) Permount, by Thermo Fisher Scientific (2 mentions) Dab detection kit, by Roche (2 mentions) Cc1 buffer, by Roche (2 mentions) Itgb4, by Vector Laboratories (2 mentions) Synaptophysin, by Vector Laboratories (2 mentions) Discovery xt processor, by Roche (2 mentions) Ezprep buffer, by Roche (2 mentions) Avidin biotin blocking, by Roche (2 mentions) Vimentin, by Vector Laboratories (2 mentions) Hnrnpl, by Fortis Life Sciences (2 mentions) Itgb1, by Cell Signaling Technology (2 mentions) Itga5, by Cell Signaling Technology (2 mentions) Cleaved caspase 3, by Cell Signaling Technology (1 mentions) Hoechst 33342, by Thermo Fisher Scientific (1 mentions) Alexa fluor 488 donkey anti rabbit igg, by Thermo Fisher Scientific (1 mentions) Alexa fluor 555 goat anti mouse igg, by Thermo Fisher Scientific (1 mentions) Odyssey fc, by LI COR (1 mentions) Snai1, by Cell Signaling Technology (1 mentions) Fibronectin, by Cell Signaling Technology (1 mentions)

6 protocols using itgb4

1

Immunohistochemical Staining of Paraffin Sections

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Immunohistochemical and immunofluorescent staining of paraffin-embedded sections was performed by using a Discovery XT processor (Ventana Medical Systems). Tissue sections were deparaffinized with EZPrep buffer (Ventana Medical Systems) and subjected to antigen retrieval with CC1 buffer (Ventana Medical Systems). Sections were blocked for 30 minutes with Background Buster solution (Innovex) and for 8 minutes with avidin-biotin blocking (Ventana Medical Systems). Sections were incubated with anti-AR (Abcam, ab133273 1 μg/mL); ITGB4 (Cell Signaling, cat# 14803, 0.5 μg/mL); Vimentin (Cell Signaling, cat# 5741, 0.5 μg/mL); and synaptophysin (Abcam, ab32127, 1 μg/mL) antibodies for 5 hours followed by biotinylated horse anti-rabbit antibodies (Vector Labs, cat# PK6101) at 1:200 dilution (for AR, ITGB4, Vimentin) or HRP-conjugated goat anti-rabbit (PI-1000) antibodies at 1:250 dilution (for synaptophysin) for 60 minutes. Detection was performed with DAB detection kit (Ventana Medical Systems) according to manufacturer instruction. Slides were counterstained with hematoxylin and mounted using Permount (Fisher Scientific).
Han H., Wang Y., Curto J., Gurrapu S., Laudato S., Rumandla A., Chakraborty G., Wang X., Chen H., Jiang Y., Kumar D., Caggiano E.G., Capogiri M., Zhang B., Ji Y., Maity S.N., Hu M., Bai S., Aparicio A.M., Efstathiou E., Logothetis C.J., Navin N., Navone N.M., Chen Y, & Giancotti F.G. (2022). Mesenchymal and stem-like prostate cancer linked to therapy-induced lineage plasticity and metastasis. Cell reports, 39(1), 110595.
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2

Immunohistochemical Staining of Paraffin Sections

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Immunohistochemical and immunofluorescent staining of paraffin-embedded sections was performed by using a Discovery XT processor (Ventana Medical Systems). Tissue sections were deparaffinized with EZPrep buffer (Ventana Medical Systems) and subjected to antigen retrieval with CC1 buffer (Ventana Medical Systems). Sections were blocked for 30 minutes with Background Buster solution (Innovex) and for 8 minutes with avidin-biotin blocking (Ventana Medical Systems). Sections were incubated with anti-AR (Abcam, ab133273 1 μg/mL); ITGB4 (Cell Signaling, cat# 14803, 0.5 μg/mL); Vimentin (Cell Signaling, cat# 5741, 0.5 μg/mL); and synaptophysin (Abcam, ab32127, 1 μg/mL) antibodies for 5 hours followed by biotinylated horse anti-rabbit antibodies (Vector Labs, cat# PK6101) at 1:200 dilution (for AR, ITGB4, Vimentin) or HRP-conjugated goat anti-rabbit (PI-1000) antibodies at 1:250 dilution (for synaptophysin) for 60 minutes. Detection was performed with DAB detection kit (Ventana Medical Systems) according to manufacturer instruction. Slides were counterstained with hematoxylin and mounted using Permount (Fisher Scientific).
Han H., Wang Y., Curto J., Gurrapu S., Laudato S., Rumandla A., Chakraborty G., Wang X., Chen H., Jiang Y., Kumar D., Caggiano E.G., Capogiri M., Zhang B., Ji Y., Maity S.N., Hu M., Bai S., Aparicio A.M., Efstathiou E., Logothetis C.J., Navin N., Navone N.M., Chen Y, & Giancotti F.G. (2022). Mesenchymal and stem-like prostate cancer linked to therapy-induced lineage plasticity and metastasis. Cell reports, 39(1), 110595.
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3

Immunostaining of Regenerated Skin

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Sectioned tissue–derived regenerated human skin or normal adult human skin was fixed with 10% formalin solution (Sigma (St. Louis, MO) HT5012) for 12 minutes. Sections were blocked (PBS, 2% bovine serum albumin, 2.5% normal goat serum, and 0.3% Triton X-100) for 30 minutes. Primary antibodies were then added to blocking buffer and put onto sections for 1 hour. The following antibodies were used at the following concentrations: FLG at 1:200 (Abcam (Waltham, MA): ab3137), LOR at 1:400 (Abcam: ab198994), KI67 (Abcam: Ab16667) at 1:300, cleaved caspase-3 at 1:300 (Cell Signaling Technology (Danvers, MA): 9661S), HNRNPL (Bethyl (Montgomery, TX): A303-895A) at 1:400, K10 (Abcam: ab9025) at 1:300, and ITGB4 (Cell Signaling Technology: 14803S) at 1:150. Secondary antibodies were used at 1:500 for 30 minutes and included Alexa Fluor 555 goat anti-mouse IgG (Thermo Fisher Scientific: A21424) and Alexa Fluor 488 donkey anti-rabbit IgG (Thermo Fisher Scientific: A21206). Hoechst 33342 (Thermo Fisher Scientific: H3570) was used at 1:1,000 to stain nuclei.
Li J., Chen Y., Tiwari M., Bansal V, & Sen G.L. (2021). Regulation of integrin and extracellular matrix genes by HNRNPL is necessary for epidermal renewal. PLoS Biology, 19(9), e3001378.
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4

Quantifying Protein Abundance with Western Blot

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Abundance of specific proteins was quantified using Western blot analysis as described (76 (link)) with the following primary antibodies: ABCG2 (Cell Signaling, 4477S, 1:250), SMAD4 (Santa Cruz, sc-7966, 1:500), phosphorylated SMAD3 (Cell Signaling, 9520S, 1:500), SMAD3 (Cell Signaling, 9523S, 1:500), fibronectin (Cell Signaling, 26836S, 1:500), SNAI1 (Cell Signaling, C15D3, 1:500), ITGB4 (Cell Signaling, 14803S, 1:500), INHBA (Abcam, ab56057, 1:500), and FST (Santa Cruz, sc365003, 1:500). The following secondary antibodies were utilized to detect bound primary antibodies: IRDye 680RD or 800CW goat anti-rabbit or mouse IgM, 1:5000, LI-COR, 926-32211, 926-68070, 926-68071. Images of Western blots probed with the fluorescent secondary antibody were collected with a LI-COR Odyssey Fc or CLx, and target proteins were quantified and normalized to total protein present. Total protein was ascertained utilizing REVERT staining. Total protein staining was used for normalization of Western blots aligning with reports demonstrating that total protein staining is a more accurate loading control than the use of housekeeping proteins (83 , 84 (link), 85 (link)). Western blots were quantified using Image studio v5.2.
Webb B.M., Bryson B.L., Williams-Medina E., Bobbitt J.R., Seachrist D.D., Anstine L.J, & Keri R.A. (2021). TGF-β/activin signaling promotes CDK7 inhibitor resistance in triple-negative breast cancer cells through upregulation of multidrug transporters. The Journal of Biological Chemistry, 297(4), 101162.
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5

Integrin and RNA Pol II Expression Analysis

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IP samples or 20 to 80 ug of cell lysates were loaded into 4% to 12% Bis-Tris (Thermo Fisher Scientific: NW04122BOX) or 3% to 8% Tris-acetate (Thermo Fisher Scientific: EA03752BOX) and transferred to PVDF membranes. Membranes were blocked in 5% BSA in TBS. Membranes were exposed to primary antibodies in blocking buffer overnight at 4 degrees. The following primary antibodies were used: ITGB4 (Cell Signaling Technology: 14803S) at 1:200, ITGB1 (Cell Signaling Technology: 9699S) at 1:250, ITGA5 (Cell Signaling Technology: 4705S) at 1:200, ITGAV (Cell Signaling Technology: 4711S) at 1:250, HNRNPL (Bethyl: A303-895A) at 1:1,000, and RNA Pol II (Active Motif (Carlsbad, CA): 91151) at 1:1,000. The loading control Beta-Actin (Santa Cruz Biotechnology (Dallas, TX) sc-47778) was used at 1:5,000. The secondary antibodies used were donkey anti-rabbit IRDye 680RD (Li-Cor 926–68073) and donkey anti-mouse IRDye 800CW (Li-Cor 926–32212) at 1:5,000.
Li J., Chen Y., Tiwari M., Bansal V, & Sen G.L. (2021). Regulation of integrin and extracellular matrix genes by HNRNPL is necessary for epidermal renewal. PLoS Biology, 19(9), e3001378.
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6

Integrin and Transcription Factor Regulation

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PLX4032, PD98059, AZD6244 and PF-573228 were purchased from Selleck Chemicals. The siRNAs against TCF4 (sc-61657), c-Myc (sc-29226), TCF3 (sc-36618) or TCF12 (sc-35552) were purchased from Santa Cruz Biotechnology. Antibodies against ITGA4, ITGA5, ITGB1, ITGB3, ITGB4, ITGB5, FAK, c-Myc, TCF12, and Porin were purchased from Cell Signaling Technology; p-FAK (Y397) antibodies were from Cell Signaling and Thermo Fisher Scientific, ID2 antibodies were from Cell Signaling, Santa Cruz Biotechnology and Thermo Fisher; V5, HMB45, COX5A, NDUFS4 and NDUFA9 antibodies were from Abcam; TCF4 antibodies were from Abnova and Santa Cruz; and PGC1α antibodies were from Santa Cruz and Millipore.
Luo C., Lim J.H., Lee Y., Granter S.R., Thomas A., Vazquez F., Widlund H.R, & Puigserver P. (2016). A PGC1α-mediated transcriptional axis suppresses melanoma metastasis. Nature, 537(7620), 422-426.
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