β actin

Manufactured by MP Biomedicals

Sourced in United States

β-actin is a cytoskeletal protein that is ubiquitously expressed in eukaryotic cells. It is a component of the microfilament system and plays a crucial role in various cellular processes, including structural support, cell motility, and intracellular transport.

Automatically generated - may contain errors

Lab products found in correlation

Close spelling variants of this product

Anti-β-actin (15 citations) Mouse anti-β-actin (10 citations) β-actin antibody (8 citations) β-actin (clone C4) (7 citations) Anti-β-actin antibody (4 citations) Antibody against β-actin (3 citations) Anti- β-Actin C4 (2 citations) β-Actin (C4, (2 citations)

31 protocols using β actin

Antibody Profiling of Subcellular Organelles

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Primary antibodies were: DLL1 (rabbit, Santa Cruz, sC-9102), Alpha 1 Sodium Potassium ATPase (mouse, Abcam, ab7671), GM130 (mouse, BD Biosciences, 610823), KDEL (mouse, Abcam, ab1223), EEA1 (rabbit, Abcam, ab2900), Rab5 (mouse, Sigma, R7904), Rab6A (mouse, Sigma, WH0005870M1), Transferrin receptor (mouse, Zymed, 13-6800), Caveolin (rabbit, BD Biosciences, 610060), Rab11 (rabbit, Sigma, R5903), Pan-Cadherin (mouse, Sigma, C1821), β-Actin (mouse, MPBiomedicals, 69100). Monoclonal antibodies against DLL1 were described [34] (link). Secondary antibodies were: Donkey anti-rat-Alexa488, goat anti-rat-Alexa488, goat anti-mouse-Alexa555, goat anti-rabbit-Alexa488, goat anti-mouse-Alexa633, goat anti-rabbit-Alexa555 (Molecular probes, Invitrogen).

Müller J., Rana N.A., Serth K., Kakuda S., Haltiwanger R.S, & Gossler A. (2014). O-fucosylation of the Notch Ligand mDLL1 by POFUT1 Is Dispensable for Ligand Function. PLoS ONE, 9(2), e88571.

+ Open protocol

+ Expand

Western Blot Analysis of Notch Signaling

Check if the same lab product or an alternative is used in the 5 most similar protocols

To obtain whole cell lysates, cells were washed with PBS and harvested in RIPA lysis buffer (G-Biosciences, St. Louis, USA) supplemented with 1× EDTA, phosphatase and protease inhibitors (Thermo Scientific). After incubation, samples were centrifuged at 4 °C for 30 min at 18,000CRF to remove cell debris. Protein concentration in the supernatant was measured using the Pierce bicinchoninic acid (BCA) assay kit (Thermo scientific). The whole cell lysates were fractionated by SDS-PAGE and transferred to a nitrocellulose membrane using a Pierce G2 Fast Blotter transfer apparatus (Thermo Scientific). Membranes were incubated with 5% milk in PBST (0.1% Tween-20) for two hours to prevent nonspecific binding of antibodies. Primary antibodies against β-actin (1:10,000) (MP Biomedicals, Santa Ana, California), Jag1, Jag2 (1:1000) (Cell Signaling Technology) were incubated overnight at 4 °C. Next, the membranes were washed three times for 5 min with PBST and incubated with 1:5000 diluted horseradish peroxidase conjugated (HRP) anti-mouse or anti-rabbit antibodies (GE Healthcare UK, Amersham, UK) for 2 h. Blots were washed three times with PBST and developed with SuperSignal West Pico Chemiluminescent Substrate (Thermo Scientific).

Kondratyev M., Pesic A., Ketela T., Stickle N., Beswick C., Shalev Z., Marastoni S., Samadian S., Dvorkin-Gheva A., Sayad A., Bashkurov M., Boasquevisque P., Datti A., Pugh T.J., Virtanen C., Moffat J., Grénman R.A., Koritzinsky M, & Wouters B.G. (2023). Identification of acquired Notch3 dependency in metastatic Head and Neck Cancer. Communications Biology, 6, 538.

+ Open protocol

+ Expand

Immunoblotting Analysis of Kinase Signaling

Check if the same lab product or an alternative is used in the 5 most similar protocols

Immunoblotting was performed as described previously [66 (link)]. The following antibodies were used: LIMK1 (BD Transduction Laboratories, #611748), PRKAR1A (BD, #610609), PRKACA/CB (BD, #610980), CFL2 (Abcam, ab39985), CFL phospho S3 (Cell Signaling, #3311). Blots were normalized to β-actin (MP Biomedicals, #69100). Quantification of bands was performed using the ImageJ software (v4.18).

Sigloch F.C., Burk U.C., Biniossek M.L., Brabletz T, & Schilling O. (2015). miR-200c dampens cancer cell migration via regulation of protein kinase a subunits. Oncotarget, 6(27), 23874-23889.

+ Open protocol

+ Expand

Comprehensive Western Blot Methodology

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Western blot studies were carried out as previously described in our laboratory.18 (link) Briefly, cells were grown to confluence and then proteins were extracted using RIPA buffer and quantified by the Bradford method. For the analysis, 50 μg of protein was resolved over 10% polyacrylamide gels and electrotransferred onto a nitrocellulose membrane. The membranes were blocked with a blocking buffer for 1 h at room temperature and then incubated overnight at 4°C with the corresponding primary antibody in blocking buffer, followed by incubation with the appropriate secondary antibody for 1 h and detection by chemiluminescence. Bands were quantified using the ImageJ photo analyzing program (US National Institutes of Health, Bethesda, ML, USA). The following primary antibodies were used: ZEB1 (Millipore, Villerica, MA, USA; Cat. ABN285), E-Cadherin (BD Transduction Laboratories, San Jose, CA, USA; Cat. 610181), N-Cadherin (Abcam, Cambridge, UK; Cat. ab12221), Vimentin (Abcam; Cat. ab8978), CK-18 (Abcam; Cat. ab7797), and β-actin as loading control (MP Bio, Santa Ana, CA, USA; Cat. 69100).

Orellana-Serradell O., Herrera D., Castellon E.A, & Contreras H.R. (2017). The transcription factor ZEB1 promotes an aggressive phenotype in prostate cancer cell lines. Asian Journal of Andrology, 20(3), 294-299.

+ Open protocol

+ Expand

Western Blot Analysis of Ube3a Knockout

Check if the same lab product or an alternative is used in the 5 most similar protocols

Western blot analysis was carried out for Ube3a^−/− and Ube3a^+/+ MEFs. The cells were washed with PBS, collected, and homogenized by sonication with an ice-cold lysis buffer containing the following (in mM): 10 HEPES pH 7.5, 150 NaCl, 50 NaF, 1 EDTA, 1 EGTA, 10 Na4P2O7 (EMD Millipore, Billerica, MA, USA), PMSF (Roche, Mannheim, Germany), and protease inhibitor cocktail (Roche, Mannheim, Germany). The samples (15 µg) were loaded on an SDS PAGE 4–20% gradient, followed by a transfer to polyvinylidene difluoride (PVDF; Roche, Mannheim, Germany) membranes and probed with primary antibodies using standard techniques. The primary antibodies and the dilutions for the Western blots were as follows: BAX 1:2000 (α-rabbit; Abcam #ab182733, Cambridge, UK), BCL-2 1:2000 (α-rabbit, Abcam #ab182858, Cambridge, UK), and UBE3A 1:1000 (α-mouse; Sigma-Aldrich #E8655, St. Louis, MO, USA). β-ACTIN 1:40,000 (α-mouse; MP Biomedicals #69100, Irvine, CA, USA) was used as a loading control. Secondary antibodies were used, respectively. The blots were developed and imaged using the Image Quant LAS 4000 system. All signals were normalized by the total protein and quantified using Image Studio Lite Ver 5.2 software.

Simchi L., Panov J., Morsy O., Feuermann Y, & Kaphzan H. (2020). Novel Insights into the Role of UBE3A in Regulating Apoptosis and Proliferation. Journal of Clinical Medicine, 9(5), 1573.

+ Open protocol

+ Expand

BRCA2, PAR, and DNA Damage Signaling

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cell lysis was performed using Mammalian Protein Extraction Reagent (MPER; Thermo Scientific), supplemented with protease inhibitor and phosphatase inhibitor (Thermo Scientific). Protein concentrations were measured using a Bradford assay. Next, proteins were separated by SDS/PAGE and transferred to polyvinylidene fluoride (PVDF; Immobilon, Merck, Burlington, MA, USA) membranes and blocked in 5% skimmed milk (Sigma) in Tris‐buffered saline (TBS) containing 0.05% Tween‐20 (Sigma). Immunodetection was performed with antibodies directed against BRCA2 (Calbiochem, Merck, Burlington, MA, USA; #OP95), PAR (Trevigen, Gaithersburg, MD, USA; #4336‐BPC‐100), phospho‐ATR (thr1898; Millipore, Burlington, MA, USA; #ABE462), STING (1 : 1000; Cell Signaling, Danvers, MA, USA, #13647), cGAS (1 : 1000; Cell Signaling; #15102S) IRF3 (1 : 1000; Cell Signaling; #4302), p‐IRF3 (1 : 100; Cell Signaling; #29047), and β‐actin (MP Biomedicals, Santa Ana, CA, USA; #69100). Horseradish peroxidase (HRP)‐conjugated secondary antibodies (DAKO, Glostrup, Denmark) were used for visualization using chemiluminescence (Lumi‐Light; Roche Diagnostics, Basel, Switzerland) on a Bio‐Rad bioluminescence device, equipped with quantity one/chemidoc xrs software (Bio‐Rad).

Schoonen P.M., Kok Y.P., Wierenga E., Bakker B., Foijer F., Spierings D.C, & van Vugt M.A. (2019). Premature mitotic entry induced by ATR inhibition potentiates olaparib inhibition‐mediated genomic instability, inflammatory signaling, and cytotoxicity in BRCA2‐deficient cancer cells. Molecular Oncology, 13(11), 2422-2440.

+ Open protocol

+ Expand

Western Blot Analysis of Metabolic Enzymes

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cell extracts were obtained from fresh plates 24 h after incubation with the corresponding growth medium. Then, cells were incubated for 30 min on ice with lysis buffer, scraped, sonicated and centrifuged at 15,000 g for 20 minutes at 4 °C. Supernatants were recovered and the protein content was quantified by the BCA kit (Pierce Biotechnology). Western blot analysis was carried out size-separating an equal amount of protein by electrophoresis on SDS polyacrylamide gels, and then the proteins were electroblotted onto polyvinylidene fluoride transfer membranes (PVDF) (Bio-Rad Laboratories, Hercules, CA, USA). The membranes were blocked with 5% of non-fat dry milk in PBS with 0.1% Tween, and then incubated with specific primary antibodies overnight at 4 °C. Next, membranes were treated with the appropriate secondary antibody for 1 hour at room temperature. All blots were visualized on Fujifilm X-ray (VWR International, Radnor, PA, USA) with chemiluminescence detection using Immobilon ECL Western Blotting Detection Kit Reagent (EMD Millipore, Billerica, MA, USA). The antibodies used were CAD (Santacruz Biotechnology), CAD-P (Cell Signaling), PDH (Merck Millipore) PDH-P (Cell signaling) and β-actin (MP Biomedicals). Also, anti-mouse (Dako) and, Anti-rabbit (Amersham Biosciences) secondary antibodies were used.

Katzir R., Polat I.H., Harel M., Katz S., Foguet C., Selivanov V.A., Sabatier P., Cascante M., Geiger T, & Ruppin E. (2019). The landscape of tiered regulation of breast cancer cell metabolism. Scientific Reports, 9, 17760.

+ Open protocol

+ Expand

Synthesis and Characterization of ILK Inhibitor T315

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

T315, an ILK inhibitor developed in the laboratory of C.S.C., was synthesized according to an established procedure,^{10 (link)} and its identity and purity were confirmed by nuclear magnetic resonance spectroscopy (300 MHz), high-resolution mass spectrometry, and elemental analysis. Stock solutions of T315 were made in dimethyl sulfoxide (DMSO) and diluted in culture medium to a final DMSO concentration of 0.1%. Antibodies against various target proteins were purchased from the following commercial sources: Akt, p-473S-Akt, FOXO3a, ILK, MLC, p-18T/19S-MLC, Mammalian target of rapamycin, p-2448S-mTOR, Snail, and ZEB1 from Cell Signaling Technology, Inc. (Danvers, MA); Twist from Abcam (Cambridge, MA); and β-actin from MP Biomedicals (Irvine CA). Control small interfering RNA (siRNA) and siRNA for ILK were purchased from Cell Signaling Technology, Inc. Protein lysates were derived from 11 thyroid cancer cell lines donated generously from the laboratories shown in Supplementary Table I. DNA was isolated from the cell lines grown in our laboratory and were then sent to Dr. C. Korch at University of Colorado on a fee-for-service basis for performing DNA fingerprinting analysis using methods described by Schweppe et al.^{12 (link)} Identity was then confirmed by comparing with DNA fingerprinting from the original clones described in the previous publication by Schweppe et al.

Shirley L.A., McCarty S., Yang M.C., Saji M., Zhang X., Phay J., Ringel M.D, & Chen C.S. (2015). Integrin-linked kinase affects signaling pathways and migration in thyroid cancer cells and is a potential therapeutic target. Surgery, 159(1), 163-170.

+ Open protocol

+ Expand

Antibody Acquisition and Characterization

Check if the same lab product or an alternative is used in the 5 most similar protocols

Antibodies were commercially acquired as follows: p-Akt (Thr308) (no. 4056; Cell Signaling Technology), p-Akt (Ser473) (no. 9271; Cell Signaling Technology), Akt (no. 2920; Cell Signaling Technology), p-4E-BP1 (Thr70) (no. 9455; Cell Signaling Technology), p-p70 S6K (Thr389) (no. 9206; Cell Signaling Technology), p70 S6K (no. 2708; Cell Signaling Technology), p-eIF2α (no. 9721; Cell Signaling Technology), β-actin (no. 691002; MP Biomedicals), 4E-BP1 (sc-6024; Santa Cruz Biotechnology), and eIF2α (sc-133132; Santa Cruz Biotechnology). Fluorophore-labeled secondary antibodies IRDye 800 and IRDye 700 were from LI-COR Biosciences.

Hatanaka M., Maier B., Sims E.K., Templin A.T., Kulkarni R.N., Evans-Molina C, & Mirmira R.G. (2014). Palmitate Induces mRNA Translation and Increases ER Protein Load in Islet β-Cells via Activation of the Mammalian Target of Rapamycin Pathway. Diabetes, 63(10), 3404-3415.

+ Open protocol

+ Expand

Western Blot Analysis of β-Catenin

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cell lysates were prepared with reporter gene lysis buffer (Promega, Mannheim, Germany) supplemented with protease inhibitor cocktail I (Calbiochem, Darmstadt, Germany). Equal amounts of protein were mixed with 2× SDS loading buffer (0.35 M Tris, pH 6.8; 30% (v/v) glycerol; 10% (w/v) SDS; 100 mM Dithiothreitol; 0.01% (w/v) bromphenol blue), separated by electrophoresis in discontinuous SDS-polyacrylamide gels and transferred to Immobilon-P membranes (Millipore, Billerica, MA, USA). Antibodies specific for β-catenin (BD Transduction Labs, San Jose, CA, USA), β-actin (MP Biomedicals, Eschwege, Germany) and the secondary horseradish peroxidase-conjugated goat anti-mouse antibody (GE Healthcare, Freiburg, Germany) were used for immuno detection. Blots were subjected to Supersignal West Dura chemiluminescence substrate (Thermo Scientific, Rockford, IL, USA) and exposed to Fuji Medical x-ray film SuperRX (FujiFilm).

Herbst A., Jurinovic V., Krebs S., Thieme S.E., Blum H., Göke B, & Kolligs F.T. (2014). Comprehensive analysis of β-catenin target genes in colorectal carcinoma cell lines with deregulated Wnt/β-catenin signaling. BMC Genomics, 15, 74.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!