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Itgb1

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ITGB1 is a laboratory reagent used in cell biology research. It is the gene that encodes the Integrin Beta 1 protein, which is a subunit of the Integrin protein complex. Integrins are cell surface receptors that mediate cell-cell and cell-extracellular matrix interactions.

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Lab products found in correlation

Gapdh, by Cell Signaling Technology (2 mentions) Sc 376638, by Santa Cruz Biotechnology (2 mentions) Ab133625, by Abcam (2 mentions) Bca kit, by Thermo Fisher Scientific (2 mentions) Ripa lysis buffer, by Merck Group (2 mentions) Ab14041, by Abcam (2 mentions) Ab68418, by Abcam (2 mentions) Ab22595, by Abcam (2 mentions) Ab22759, by Abcam (2 mentions) Ab92726, by Abcam (2 mentions) Thbs1, by Cell Signaling Technology (2 mentions) Sparc, by Cell Signaling Technology (2 mentions) Ab3269p, by Merck Group (2 mentions) Itgb3, by Cell Signaling Technology (2 mentions) Itga5, by Cell Signaling Technology (2 mentions) Itgb4, by Cell Signaling Technology (2 mentions) Ecl reagent, by Cytiva (1 mentions) N cadherin, by Cell Signaling Technology (1 mentions) Pf573228, by Merck Group (1 mentions) P ndrg1 t346, by Cell Signaling Technology (1 mentions)

Spelling variants of this product

Anti-ITGB1 (7 citations) Anti-ITGB1 antibodies (3 citations)

11 protocols using itgb1

1

Western Blot Analysis of EMT Markers

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Proteins in clinical tissues and HCC cells were extracted using RIPA buffer and subjected to concentration measurements using BCA kit. After being separated in SDS-PAGE gels, the protein (20 µg) on SDS-PAGE gels (4–20%) were transferred to polyvinylidene fluoride membrane. These membranes were incubated with 5% non-fat dry milk (diluted in TBST) at room temperature for 1 h and primary antibodies of E-cadherin (dilution 1:1,000; cat. no. 3195; Cell Signaling Technologies, Inc., Danvers, MA, USA), N-cadherin (dilution 1:500; cat. no. 4061; Cell Signaling Technologies, Inc.), ITGB1 (dilution 1:1,000; cat. no. 4706; Cell Signaling Technologies, Inc.) and GAPDH (dilution 1:2,000; cat. no. sc-32233; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) overnight at 4°C. GAPDH was used as internal control. Protein signals were detected using ECL reagents (Amersham Biosciences; GE Healthcare, Chicago, IL, USA).
Zhang L., Zhang T., Deng Z, & Sun L. (2019). MicroRNA-3653 inhibits the growth and metastasis of hepatocellular carcinoma by inhibiting ITGB1. Oncology Reports, 41(3), 1669-1677.
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2

Pharmacological Modulation of PI3K/AKT Pathway

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LY294002 (Cell Signaling, #9901) and Bapta-AM (Selleckchem, #S7534) were used at a 50 μM concentration. PP2 (Cayman Chemical, #13198–1), U0126 (Cell signaling, #9903), PF-573228 (Sigma, #PZ0117), Gefitinib (Invivogen, #tlrl-gef) were used at 10 μM concentration. Thapsigargin (Cayman Chemical, #10522) was used at 5 μM concentration. Wotrmannin (Cell Signaling, #9951) and MK-2206 2HCl (Selleckchem, #S1078) were used at 1 μM concentration.
Crystal violet was purchased from Sigma (#3886).
Antibodies for AKT (#4691), p-AKT-S473 (#4060), p-NDRG1-T346 (#5482), NDRG1 (#9485), p-P70S6K-T389 (#9234), P70S6K (#2708), RICTOR (#2114), p-PRAS40-T246 (#2997), PRAS40 (#2691), TSC2 (#4308), p-TSC2-T1462 (#3617), 4EBP1 (#9644), p-4EBP1-T37/46 (#2855), ITGB1 (#9699), ILK (#3856), SRC (#2123), YES (#3201), were from Cell Signaling. Antibody against β-Actin (#A3854) was from Sigma.
Merhi A., Delrée P, & Marini A.M. (2017). The metabolic waste ammonium regulates mTORC2 and mTORC1 signaling. Scientific Reports, 7, 44602.
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3

Quantitative Protein Analysis of sEVs

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sEV and cells were resuspended in RIPA lysis buffer (Millipore-Sigma) and incubated on ice for 20 min prior to centrifugation at 12,000 g for 10 min. Supernatants were used for western blotting in SDS-PAGE electrophoresis. Protein concentration was determined by a BCA kit (Thermofisher). The following antibodies were used: ADIPOQ (AB3269P, Millipore-Sigma), CD36 (ab133625, Abcam), HSP90alpha (PA3–013, Thermofisher), POSTN (ab14041, Abcam), PGAM1/4 (sc-376638, Santa Cruz); THBS1 (14778, Cell Signaling), ITGB1 (4706, Cell Signaling), EGFR (2232, Cell Signaling), SPARC (8725, Cell Signaling), IGFBP5 (AF578, R&D systems), CD9 (ab92726, Abcam), CD63 (ab68418, Abcam), CANX (ab22595, Abcam) and FASN (ab22759, Abcam). Given the large difference in the abundance for some of the proteins among different cell types, some of the blots were overexposed to show the signal in as many cell types as possible. Because of this, in some cases, the signal is beyond the linear range, and thus the quantification might not be absolutely accurate. Band intensity quantification was performed using ImageJ software (NIH).
Garcia-Martin R., Brandao B.B., Thomou T., Altindis E, & Ronald Kahn C. (2022). Tissue differences in the exosomal/small extracellular vesicle proteome and their potential as indicators of altered tissue metabolism. Cell reports, 38(3), 110277.
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4

Immunohistochemical Analysis of Lung Adenocarcinoma

Cited 1 time
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Microarrays of human lung adenocarcinoma tissues (HLugA150CS03 and HlugA180Su04) were obtained from Shanghai Outdo Biotech Co., Ltd. (Shanghai, China). Samples underwent USP51, PD‐L1 (17952‐1‐AP, Proteintech, Wuhan, Hubei, China) and ITGB1 (34971T, Cell Signaling Technology) antibody staining for IHC assays with the Envision Kit (Dako) in accordance as per kit protocols. Several pathologists then blindly scored the immunostaining, as described previously [24 (link)]. This protocol received ethical approval from the Nankai University (NKUIRB2021010).
Li J., Xiao X., Ou Y., Cao L., Guo M., Qi C., Wang Z., Liu Y., Shuai Q., Wang H., Sun P., Shi Y., Yang G, & Yang S. (2023). USP51/PD‐L1/ITGB1‐deployed juxtacrine interaction plays a cell‐intrinsic role in promoting chemoresistant phenotypes in non‐small cell lung cancer. Cancer Communications, 43(7), 765-787.
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5

Immunohistochemical Staining of Cellular Markers

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Following deparaffinization and rehydration, epitopes were retrieved by boiling slides for 10 min at 100 C in sodium citrate buffer (10 mM sodium citrate, 0.05% Tween-20, pH 6.0). Slides were incubated for 1 h at room temperature in primary antibody (Rab11, Abcam ab3612; Rab11b, Thermo-Fisher PA5-31348; K8, Abcam ab53280; Ki-67, Cell Signaling Technology 9027; ITGB1, Cell Signaling Technology 9699). Primary antibodies were detected using the VECTASTAIN Elite ABC HRP Kit (Vector Laboratories), followed by detection with ImmPACT DAB (Vector Laboratories) following manufacturer’s directions. Images were taken with an Olympus BX43.
Howe E.N., Burnette M.D., Justice M.E., Schnepp P.M., Hedrick V., Clancy J.W., Guldner I.H., Lamere A.T., Li J., Aryal U.K., D’Souza-Schorey C., Zartman J.J, & Zhang S. (2020). Rab11b-mediated integrin recycling promotes brain metastatic adaptation and outgrowth. Nature Communications, 11, 3017.
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6

Protein Extraction and Western Blot Analysis

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Total proteins from GC tissues and cells were extracted using radio immunoprecipitation assay (RIPA) lysis buffer (Beyotime, Shanghai, China) containing the protease inhibitors and phosphatase inhibitors. Protein samples (20 μL) in each group were separated by SDS-PAGE before being transferred onto polyvinylidene fluoride (PVDF) membranes (Beyotime, Shanghai, China), which were then blocked by 5% non-fat milk followed by incubation overnight at 4°C with primary antibodies against ITGB1 (Cell Signaling Technology, Beverly, MA, USA; #34971), FAK (Cell Signaling Technology, Beverly, MA, USA; #3285), pY397-FAK (Cell Signaling Technology, Beverly, MA, USA; #3281), CDC42 (Abcam, Cambridge, MA, USA; #ab187643), PXN (Abcam, Cambridge, MA, USA; #ab32084), pY118-PXN (Abcam, Cambridge, MA, USA; #ab109547), AKT (Proteintech, Wuhan, China; #10176-2-AP), pS473-AKT (Proteintech, Wuhan, China; #66444-1-IG), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Santa Cruz Biotechnology, Dallas, TX, USA; #sc-47724) (Table S4). Subsequently, the membranes were incubated with horseradish peroxidase-labeled secondary antibody (Proteintech, Wuhan, China) at room temperature for 1 h. Protein signals were developed using the Enhanced Chemiluminescence Kit (Beyotime, Shanghai, China).
Zhang L., Yao L., Zhou W., Tian J., Ruan B., Lu Z., Deng Y., Li Q., Zeng Z., Yang D., Shang R., Xu M., Zhang M., Cheng D., Yang Y., Ding Q, & Yu H. (2021). miR-497 defect contributes to gastric cancer tumorigenesis and progression via regulating CDC42/ITGB1/FAK/PXN/AKT signaling. Molecular Therapy. Nucleic Acids, 25, 567-577.
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7

Quantitative Protein Analysis of sEVs

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sEV and cells were resuspended in RIPA lysis buffer (Millipore-Sigma) and incubated on ice for 20 min prior to centrifugation at 12,000 g for 10 min. Supernatants were used for western blotting in SDS-PAGE electrophoresis. Protein concentration was determined by a BCA kit (Thermofisher). The following antibodies were used: ADIPOQ (AB3269P, Millipore-Sigma), CD36 (ab133625, Abcam), HSP90alpha (PA3–013, Thermofisher), POSTN (ab14041, Abcam), PGAM1/4 (sc-376638, Santa Cruz); THBS1 (14778, Cell Signaling), ITGB1 (4706, Cell Signaling), EGFR (2232, Cell Signaling), SPARC (8725, Cell Signaling), IGFBP5 (AF578, R&D systems), CD9 (ab92726, Abcam), CD63 (ab68418, Abcam), CANX (ab22595, Abcam) and FASN (ab22759, Abcam). Given the large difference in the abundance for some of the proteins among different cell types, some of the blots were overexposed to show the signal in as many cell types as possible. Because of this, in some cases, the signal is beyond the linear range, and thus the quantification might not be absolutely accurate. Band intensity quantification was performed using ImageJ software (NIH).
Garcia-Martin R., Brandao B.B., Thomou T., Altindis E, & Ronald Kahn C. (2022). Tissue differences in the exosomal/small extracellular vesicle proteome and their potential as indicators of altered tissue metabolism. Cell reports, 38(3), 110277.
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8

Bone Protein Expression Analysis in Col-OMT Mice

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The femora of control and Col-OMT male mice were removed from soft tissue, and the bone marrow was flushed away. Femora were frozen in liquid nitrogen and then crushed with a mortar and pestle. Then, 10 µg of protein from either bone tissue or cell lysates was separated by 10% SDS-PAGE and transferred to PVDF membranes. After blocking with 5% skim milk, the membrane was probed with a primary antibody against Osx (Santa Cruz Biotechnology), Itgb3 (Cell Signaling Technology, Beverly, MA, USA), Itgb1 (Cell Signaling Technology), p-vinculin (Abcam), GAPDH (Bioworld Technology, St Louis Park, MN, USA), and HSP90 (Enzo Life Sciences, Farmingdale, NY, USA). A horseradish peroxidase-conjugated goat anti-rabbit IgG (Enzo Life Sciences) secondary antibody was used for visualization. Signals were detected using a Las-4000 imager (GE Healthcare Life Science, Pittsburgh, PA, USA).
Moon Y.J., Yun C.Y., Choi H., Kim J.R., Park B.H, & Cho E.S. (2018). Osterix regulates corticalization for longitudinal bone growth via integrin β3 expression. Experimental & Molecular Medicine, 50(7), 1-11.
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9

Quantitative Protein Analysis by Western Blot

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Protein quantification by Western blot was performed as we previously described (Chang et al., 2017 (link)). Briefly, total proteins were extracted and separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and then transferred onto PVDF membrane (Millipore). After incubation with primary antibodies against ITGB1 (#9699, 1;1000, Cell Signaling Technology, Danvers, MA, United States), EZH2 (17-662, 1;1000, Millipore), MYC (#18583, 1;1000, Cell Signaling Technology), β-catenin (#8480, 1;1000, Cell Signaling Technology), histone H3 (#14269, 1;1000, Cell Signaling Technology), or GAPDH (#5174, 1;1000, Cell Signaling Technology), the membranes were further incubated with Goat anti-Rabbit or Goat anti-Mouse secondary antibodies.
Chang R., Xiao X., Fu Y., Zhang C., Zhu X, & Gao Y. (2021). ITGB1-DT Facilitates Lung Adenocarcinoma Progression via Forming a Positive Feedback Loop With ITGB1/Wnt/β-Catenin/MYC. Frontiers in Cell and Developmental Biology, 9, 631259.
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10

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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