Ab183666

Manufactured by Abcam

Sourced in United Kingdom

Ab183666 is an antibody product offered by Abcam. It is a primary antibody used for research applications. The core function of this product is to serve as a detection reagent for target proteins in various experimental techniques.

Automatically generated - may contain errors

Lab products found in correlation

Image pro plus 6.0 program, by Media Cybernetics (1 mentions) Ab2413, by Abcam (1 mentions) Alexa fluor 594 goat anti rabbit igg, by Thermo Fisher Scientific (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions) Leica tcs sp5 confocal microscope, by Leica camera (1 mentions) Phosphate buffered saline (pbs), by Merck Group (1 mentions) Ab24170, by Abcam (1 mentions) Na934v, by GE Healthcare (1 mentions) Na931v, by GE Healthcare (1 mentions) Na935, by GE Healthcare (1 mentions) Anti myc 4a6, by Merck Group (1 mentions) Ahp500gt, by Bio-Rad (1 mentions) Anti gapdh 6c5, by Merck Group (1 mentions) Anti lamin b sc 6216, by Santa Cruz Biotechnology (1 mentions) Hoechst 33342, by Dojindo Laboratories (1 mentions) Alexa fluor 488, by Thermo Fisher Scientific (1 mentions) Alexa fluor 594, by Thermo Fisher Scientific (1 mentions) Ab13370, by Cell Signaling Technology (1 mentions) P akt ser473, by Cell Signaling Technology (1 mentions) Arg56418, by Arigo Biolaboratories (1 mentions)

10 protocols using ab183666

Fluorescent Immunohistochemistry of Fibronectin and Integrin β1

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For fluorescent immunohistochemistry (IHC) of fibronectin and integrin β1,
deparaffinized slides were subjected to antigen retrieval in boiled sodium
citrate buffer solution (pH 6.0) for 10 min, then blocked for 20 min in blocking
buffer (5% bovine serum albumin and 0.01% Triton X-100 in phosphate buffer
solution (PBS; Sigma). After blocking, samples were incubated with rabbit
polyclonal anti-fibronectin (Abcam, ab2413, 1:300) and rabbit polyclonal
anti-integrin β1 (Abcam, ab183666, 1:300) overnight at 4°C. Alexa Fluor 594 goat
anti-rabbit IgG (Life Technologies, USA, 1:500) were used to detect primary
antibodies, and then counterstained with 4′,6-diamidino-2-phenylindole (DAPI;
Invitrogen). Immunofluorescent images were captured using a Leica TCS SP5
confocal microscope (Leica). The expression intensity was measured using the
Image-Pro Plus 6.0 program (Media Cybernetics, Rockville, MD, USA).

Liu Y., Wang H., Dou H., Tian B., Li L., Jin L., Zhang Z, & Hu L. (2020). Bone regeneration capacities of alveolar bone mesenchymal stem cells sheet in rabbit calvarial bone defect. Journal of Tissue Engineering, 11, 2041731420930379.

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Antibody-based Protein Localization Assay

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The following antibodies and reagents were used: anti-LAMP1 (ab24170, abcam), anti-LAMP2 (clone H4B4, Developmental Studies Hybridoma Bank), anti-Lamin B (sc-6216, Santa Cruz), anti-Rab11 (clone 47, BD Biosciences), anti-GAPDH (6C5, EMD Millipore), anti-DYKDDDDK (clone L5, Novus Biologicals), anti-myc (4A6, Merck Millipore) (562, MBL Life Science), anti-TGN46 (AHP500GT, Bio-Rad), anti-integrin beta 1 (ab183666, abcam), anti-EEA1 (610456, BD Biosciences), anti-Transferrin receptor (3C11F11, proteintech), anti-mouse IgG HRP linked whole antibody from sheep (NA931V, GE Healthcare), anti-rabbit IgG HRP linked whole antibody from donkey (NA934V, GE Healthcare), anti-rat IgG HRP linked whole antibody from goat (NA935, GE Healthcare), Hoechst 33342 (Dojindo), donkey anti-mouse IgG (H+L) highly cross-adsorbed secondary antibody, Alexa Fluor 594 (A-21203, Invitrogen), donkey anti-rabbit IgG (H+L) highly cross-adsorbed secondary antibody, Alexa Fluor 488 (A-11008, Invitrogen), donkey anti-rat IgG (H+L) highly cross-adsorbed secondary antibody, Alexa Fluor 488 (A-21208, Invitrogen), Donkey anti-Sheep IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 594 (A-11016, Invitrogen). Biotinylated PhoSL was kindly provided by J Chemical (58 (link)), and other biotinylated lectins were purchased from J Chemical (ConA, #J203; L4-PHA, #J212; SSA, #J218; MAM, #J210).

Kitano M., Kizuka Y., Sobajima T., Nakano M., Nakajima K., Misaki R., Itoyama S., Harada Y., Harada A., Miyoshi E, & Taniguchi N. (2021). Rab11-mediated post-Golgi transport of the sialyltransferase ST3GAL4 suggests a new mechanism for regulating glycosylation. The Journal of Biological Chemistry, 296, 100354.

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Western Blot Analysis of Cellular Signaling Proteins

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The cells were lysed using RIPA lysis buffer (P0013B; Beyotime Institute of Biotechnology), containing a protease and phosphatase inhibitor cocktail. Lysate protein concentrations were determined using a BCA kit (P0010S; Beyotime Institute of Biotechnology). The denatured protein samples were separated by 10% SDS-PAGE in electrophoresis buffer and transferred to a PVDF membrane. The membranes were blocked in 5% milk for 2 h, then incubated with primary antibodies overnight at 4°C. The antibodies were directed against Rap1A (rabbit polyclonal, ab197673, 1:1,000; Abcam), β1 integrin (rabbit polyclonal, ab183666, 1:3,000; Abcam), matrix metalloproteinase (MMP)9 (rabbit polyclonal, 10375-2-AP, 1:1,000; ProteinTech Group, Inc.), E-cadherin (rabbit monoclonal, 24E10, 1:1,000; Cell Signaling Technology, Inc.), Slug (rabbit polyclonal, arg55242, 1:1,000; Abcam), AKT (rabbit polyclonal, arg56418, 1:600; Arigo), SP1 (rabbit polyclonal, ab13370, 1:3,000) and pAKTser473 (rabbit monoclonal, 4060, 1:2,000; Cell Signaling Technology, Inc.). The membranes were incubated with HRP-conjugated secondary antibodies for 2 h, and then antigen-antibody complexes were detected using an electrochemiluminescence kit to compare the corresponding protein expression levels among different samples.

Li Q., Xu A., Chu Y., Chen T., Li H., Yao L., Zhou P, & Xu M. (2019). Rap1A promotes esophageal squamous cell carcinoma metastasis through the AKT signaling pathway. Oncology Reports, 42(5), 1815-1824.

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Profiling Inflammatory Signaling in Liver

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Total liver tissue homogenates and whole cell lysates were used for Western blotting to determine (i) protein levels of proinflammation cytokines: TNF-α (Proteintech, 60291-1-Ig, 1:1000) and IL-1β (Abcam, ab234437, 1:1000); (ii) the activation state of NF-κB p50/p65 signals including NF-κB p105 (Proteintech, 66992-1-Ig, 1:1000), p-NF-κB p50 (Abclonal, AP0125, 1:1000)/NF-κB p50 (Proteintech, 66992-1-Ig, 1:1000), p-NF-κB p65 (Affinity,AF3388, 1:2000)/NF-κB p65 (Abclonal, A19653, 1:500), and p-IκB (Affinity,AF2002, 1:2000)/IκB (Affinity,AF5002, 1:2000); and (iii) protein levels of Sema7a (Proteintech, 67397-1-Ig, 1:1000) and integrin β1 (Abcam, ab183666, 1:5000). Quantification of Western blotting results was determined by densitometric scanning using ImageJ software. Raw data of Western blotting results were included in Additional file 4.

Li X., Xie W., Pan Q., Zhang X., Zhang L., Zhao N., Xie Q., Ding J, & Chai J. (2023). Semaphorin 7A interacts with nuclear factor NF-kappa-B p105 via integrin β1 and mediates inflammation. Cell Communication and Signaling : CCS, 21, 24.

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β1 Integrin Immunoprecipitation and Immunoblotting

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For β1 integrin immunoprecipitation, cells were lysed in NP-40 buffer containing 50 mM Tris-HCl (pH 7.5), 150 mM NaCl, 0.3% Nonidet P-40, and protease inhibitor cocktail (Sigma-Aldrich). Cell lysates (500 μl) were incubated with anti-β1 integrin antibody (Abcam #ab183666) or control IgG and Protein A/G PLUS-Agarose (40 μl, Santa Cruz Biotechnology, sc-2003) overnight at 4 °C. The beads were washed three times with NP-40 buffer, followed by immunoblotting. Immunoblotting was performed following standard protocol. Antibodies were purchased from the indicated companies, including Cell Signaling (PTEN, #9188S; S473P-AKT, #3787S; AKT, #2910S; T416P-SRC, #6943S; SRC, #2109S; Notch1, #4380S; YAP1, #14074S; T402P-PYK2, #3291S; PYK2, #3090S; T397P-FAK, #3283S;FAK, #3285S; T1068P-EGFR, #3777S and EGFRviii, #64952S), Abcam (LOX, #ab174316; CXCL5, #ab9802; SEMA3E, #ab128619, SERPINE2, #ab134905 and β1 integrin, #ab179471), R&D system (FABP5, #AF3077), EMD Millipore (Vinculin, #05–386), Santa Cruz (P53, #sc-126) and Sigma (ȕ-actin, #A3854).

Chen P., Zhao D., Li J., Liang X., Li J., Chang A., Henry V.K., Lan Z., Spring D.J., Rao G., Wang Y.A, & DePinho R.A. (2019). Symbiotic macrophage-glioma cell interactions reveal synthetic lethality in PTEN null glioma. Cancer cell, 35(6), 868-884.e6.

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Western Blot of Cell Adhesion Proteins

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Western blot was performed as previously described [32 (link)]. The following antibodies were used: anti-GAPDH (ab9485; Abcam), anti-paxillin (610568, BD), anti-FAK (610087, BD), anti-p-FAK (Tyr397) (44-624G, Thermofisher), anti-p-paxillin (Tyr118) (44-722G, Thermofisher), anti-p-paxillin (44-720G, Thermofisher), anti-Rac1 (507720, Zenbio), anti-VE-Cadherin (ab33168; Abcam), anti-integrin α5 (ab6131; Abcam), anti-integrin β1 (ab183666; Abcam), anti-integrin β3 (ab197662; Abcam), anti-vinculin (ab129002; Abcam). The secondary antibody was HRP-labeled IgG (A0208, A0216; Beyotime).

Guo Y., Mei F., Huang Y., Ma S., Wei Y., Zhang X., Xu M., He Y., Heng B.C., Chen L, & Deng X. (2021). Matrix stiffness modulates tip cell formation through the p-PXN-Rac1-YAP signaling axis. Bioactive Materials, 7, 364-376.

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Western Blot Analysis of Tumor Tissue

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Tumor tissues were lysed in 5% Triton X-100 buffer followed by centrifugation (20,000×g, 4 °C, 15 minutes) and collecting of supernatants. Equal amounts of protein (10 μg) were mixed with loading dye, boiled for 5 min, separated on a denaturing SDS-polyacrylamide gel and transferred to a PVDF membrane. The membrane was blocked in 5% dry milk or BSA in TBS buffer with 0.1% Tween (TBS-T) for 1 hour and incubated overnight with antibodies against α-smooth muscle actin (ab7817, Abcam), transforming growth factor β (3711, Cell Signaling), β1-integrins (ab183666, Abcam), phosphorylated AKT1 (ab81283, Abcam), or β-actin (4967, Cell Signaling) as a reference. Then, the membrane was washed twice with TBS-T and incubated with HRP-conjugated secondary antibody (ab6721, Abcam or sc-2005, Santa Cruz Biotechnology) at room temperature for 1 h, followed by several washings with TBS-T and deionized water. Protein bands were visualized by ChemiDoc XRS+ imaging system (Bio-Rad Laboratories, Hercules, CA) using chemiluminescence mode.

Durymanov M., Kroll C., Permyakova A., O'Neill E., Sulaiman R., Person M, & Reineke J. (2018). Subcutaneous Inoculation of 3D Pancreatic Cancer Spheroids Results in Development of Reproducible Stroma-Rich Tumors. Translational Oncology, 12(1), 180-189.

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Multiprotein Immunofluorescence Staining

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The samples were rinsed with PBS and fixed for 10 min in 4 % (w v⁻¹) paraformaldehyde, permeabilized for 10 min in 0.2 % (v v⁻¹) Triton X-100, and blocked for 1 h in 5 % (w v⁻¹) BSA at room temperature. Then, the samples were incubated with the following primary antibodies overnight at 4 °C: anti-HtrA3 (A14649; ABclonal), anti-YAP1 (14074S; Cell Signaling Technology), anti-RUNX2 (12556; Cell Signaling Technology), anti-BMP2 (ab284387; Abcam), and anti-integrin β1 (ab183666; Abcam) antibodies. After washing three times in PBS, the samples were probed with the secondary antibodies, Alexa Fluor 594 (ab150080; Abcam), for 1 h in the dark. The cell nuclei were stained with DAPI. Images were acquired using a confocal microscope.

Guo Y., Ma S., Wang D., Mei F., Guo Y., Heng B.C., Zhang S., Huang Y., Wei Y., He Y., Liu W., Xu M., Zhang X., Chen L, & Deng X. (2024). HtrA3 paves the way for MSC migration and promotes osteogenesis. Bioactive Materials, 38, 399-410.

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Extracellular Vesicle Protein Marker Analysis

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Primary antibodies were: anti-ALIX(1:1000), ab186429 (Abcam); anti-TSG101(1:1000), GTX70255 (GeneTex); anti-flotillin1(FLOT)(1:1000), 610820 (BD); anti-integrin β 1(1:1000), ab183666 (Abcam); ant-CD9(1:1000), ab223052 (Abcam); anti-Annexin A5(1:1000), #8555 S (CST); anti-Annexin A2(1:7000)(ANXA2), ab178677 (Abcam); anti-VWF(1:1000), 65707 (CST); anti-INSR(1:200), sc57342 (Santa Cruz Biotechnology); anti-Calnexin(1:1000), ab22595 (Abcam); anti-TOM20(1:1000), 11802-1-AP (Proteintech); anti-β-actin(1:1000), 8457 S (CST).

Tian W., Shi D., Zhang Y., Wang H., Tang H., Han Z., Wong C.C., Cui L., Zheng J, & Chen Y. (2024). Deep proteomic analysis of obstetric antiphospholipid syndrome by DIA-MS of extracellular vesicle enriched fractions. Communications Biology, 7, 99.

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Western Blot Analysis of Cell Adhesion and Muscle Proteins

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After protein quantization, 15 μg of protein was loaded, resolved on 10% sodium dodecyl sulfate (SDS)-polyacrylamide gels, electrotransferred onto nitrocellulose membranes, and blocked with serum. The membranes were incubated overnight with rabbit antibodies against ITGA1 (1:1000 dilution; ab181434; Abcam, Cambridge, UK), rabbit antibodies against ITGB1 (1:5000 dilution; ab183666; Abcam, Cambridge, UK), rabbit antibodies against LAMC1 (1:1000 dilution; ab233389; Abcam, Cambridge, UK), and rabbit antibodies against CKM (1:1000 dilution; sc-365046; Santa Cruz Biotechnology, Inc., Dallas, TX, USA). After 3 times of washing, secondary antibodies were added. Blots were detected by an enhanced chemiluminescence (ECL) reagent (ECL-Plus/Kit; Amersham, Piscataway, NJ, USA). Besides, β-actin (1:1000 dilution; Sigma-Aldrich, St. Louis, MO, USA) was used as loading control.

Li Y., Zhang D., Jin B., Xia L, & Zhang A. (2020). Proteomic Analysis of Uterine Tissues During Peri-Implantation Period in Mice with Experimentally Induced Adenomyosis that Treated with anti-Ngf: Implications for Cell-Cell Adhesion and Metabolic Processes. Reproductive Sciences, 28(1), 207-217.

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