Actin antibody

Manufactured by Abcam

Sourced in United States

Actin antibody is a primary antibody that binds to and detects actin, a highly conserved protein found in all eukaryotic cells. Actin is a key component of the cytoskeleton and is involved in various cellular processes such as cell motility, cell division, and muscle contraction.

Automatically generated - may contain errors

Lab products found in correlation

Anti adenovirus type 5 antibody, by Abcam (2 mentions) Electrochemiluminescence reagent, by Cytiva (1 mentions) Horseradish peroxidase, by Santa Cruz Biotechnology (1 mentions) Anti rabbit igg antibody, by Santa Cruz Biotechnology (1 mentions) Anti mouse secondary antibody, by Vector Laboratories (1 mentions) Anti rabbit secondary antibody, by Vector Laboratories (1 mentions) Dkk3 antibody, by Santa Cruz Biotechnology (1 mentions) Smad4 antibody, by Abcam (1 mentions) Goat anti rabbit secondary antibody, by Vector Laboratories (1 mentions) Fitc conjugated pna, by Merck Group (1 mentions) Alexa fluor 488 or 568 conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions) Anti β 3 tubulin, by R&D Systems (1 mentions) Anti ki67, by Abcam (1 mentions) Bca protein assay kit, by Thermo Fisher Scientific (1 mentions) Protease and phosphatase inhibitor, by Thermo Fisher Scientific (1 mentions) Cytokeratin 14, by Abcam (1 mentions) Immobilon fl pvdf membrane, by Merck Group (1 mentions) Nupage 10 bis tris gel, by Thermo Fisher Scientific (1 mentions) Egf receptor, by Cell Signaling Technology (1 mentions) Ripa lysis buffer, by Thermo Fisher Scientific (1 mentions)

14 protocols using actin antibody

Western Blot Analysis of ADK Protein Expression

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

An equal amount of proteins was loaded on 12% sodium
dodecyl sulfate-polyacrylamide gel electrophoresis (SDSPAGE)
and transferred to polyvinylidene fluoride membranes.
Blots were blocked using non-fat dry milk for 1 hour. After
blocking, membranes were probed with ADK primary
antibody (Santa Cruz, 1/1000). Then anti-rabbit IgG antibody
conjugated with horseradish peroxidase (Santa Cruz, 1/8000)
was used for 1 hour at room temperature. Subsequently, the
blots were reprobed with a ß-actin antibody (1:2000, Abcam,
ab8227, USA). Then, the blots were treated with Electro
Chemi Luminescence reagents (Amersham Biosciences,
UK). For quantification of protein intensities, western blot
bands were visualized on radiograph film and evaluated by
Image J software. Human umbilical cord tissue was used in
accordance with the Declarations of Tehran University of
Medical Science and Stem Cell Research Center Committee
after obtaining written informed consent from the mother.

Estiri H., Fallah A., Soleimani M., Aliaghaei A., Karimzadeh F., Babaei Abraki S, & Ghahremani M.H. (2018). Stable Knockdown of Adenosine Kinase by Lentiviral Anti-ADK miR-shRNAs in Wharton’s Jelly Stem Cells. Cell Journal (Yakhteh), 20(1), 1-9.

+ Open protocol

+ Expand

Western Blot Analysis of DKK3 and SMAD4

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

Western was performed using a 1:1000 dilution of DKK3 antibody (Santa Cruz) and a 1:5000 dilution of goat anti-rabbit secondary antibody (Vector). While the calculated molecular weight of DKK3 is 38 kDa, the size on Western has been reported as 50-55 kDa in reducing conditions due to glycosylation.^{13 (link)} For SMAD4, a 1:2000 dilution of SMAD4 antibody (Abcam) was used with a 1:8000 dilution of anti-rabbit secondary antibody (Vector). ß-actin was used as a loading control with a 1:10000 dilution of ß-actin antibody (Abcam) and a 1:10000 dilution of anti-mouse secondary antibody (Vector).

Wang Z., Lin L., Thomas D.G., Nadal E., Chang A.C., Beer D.G, & Lin J. (2015). The Role of Dickkopf-3 Overexpression in Esophageal Adenocarcinoma. The Journal of thoracic and cardiovascular surgery, 150(2), 377-385.e2.

+ Open protocol

+ Expand

Immunofluorescence and Western Blot Analysis of SMGs

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

The cultured SMGs were fixed with 4% paraformaldehyde (PFA) and incubated with fluorescein isothiocyanate (FITC)-conjugated PNA (1:200, Sigma-Aldrich, MO), anti-βIII-tubulin (1:1000, R&D Systems), anti-FGF7 (1:1000, AB Biotech), anti-FGF10 (1:2000, Millipore), or anti-ki-67 (1:1500, Abcam, UK). The antibodies were detected using Alexa Fluor 488 or 568 conjugated secondary antibodies (Life Technologies). Images were obtained using confocal microscopy (C1, Nikon). The total amount of protein obtained from the cultured tissue was calculated using a BCA protein assay kit (Pierce Biotechnology, IL) and then the following antibodies were used in Western blot analysis: anti-FGF7 (1:1000) and anti-FGF10 (1:2000). Goat anti-rabbit horseradish peroxidase conjugated secondary antibody (1:8000, Santa Cruz Biotec, CA) was used as a secondary antibody. ß-Actin antibody (1:5000, Abcam) was used as a loading control. Western blot data were quantified by comparing the relative density of target bands to the loading control by using Image J software. Additional information is supplied in the Supplementary data.

Taketa H., Sathi G.A., Farahat M., Rahman K.A., Sakai T., Hirano Y., Kuboki T., Torii Y, & Matsumoto T. (2015). Peptide-modified Substrate for Modulating Gland Tissue Growth and Morphology In Vitro. Scientific Reports, 5, 11468.

+ Open protocol

+ Expand

Antibody Characterization for Adenovirus

Cited 2 times

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

Mouse monoclonal anti-E1A antibodies, previously described [11 (link),12 (link)], and as used by us earlier [9 (link),10 (link)], were grown in-house and used as the supernatant from the hybridoma cells. Mouse monoclonal anti-72kDa DBP antibody was previously described [13 (link)] and was used at a dilution of 1:400 for Western blot. Anti-adenovirus type 5 antibody was purchased from Abcam (catalog number ab6982, Cambridge, MA, USA). Actin antibody was purchased from Abcam (catalog number ab3280). Secondary antibodies were acquired from Jackson ImmunoResearch (West Grove, PA, USA) and were used at a dilution of 1:200,000.

Costa R., Akkerman N., Graves D., Crisostomo L., Bachus S, & Pelka P. (2020). Characterization of Adenovirus 5 E1A Exon 1 Deletion Mutants in the Viral Replicative Cycle. Viruses, 12(2), 213.

+ Open protocol

+ Expand

Antibody Detection Assay for Adenovirus E1A

Cited 1 time

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

As performed previously [2 (link)], mouse monoclonal anti-E1A M58 antibody was previously described [13 (link)] and was grown in-house and used as the supernatant of M58 hybridoma cells. The M58 antibody binds within the region encoded by the first exon of E1A (between residues 63 and 99 of E1A) and recognizes all mutant E1A proteins used in this study [14 (link)]. Mouse monoclonal anti-72k DNA binding protein (DBP) antibody was previously described [15 (link)] and was used at a dilution of 1:400 for western blot. Anti-adenovirus type 5 antibody was purchased from Abcam (catalog number ab6982, Cambridge, MA, USA). Actin antibody was purchased from Abcam (catalog number ab3280). Secondary antibodies were acquired from Jackson ImmunoResearch (West Grove, PA, USA) and were used at a dilution of 1:200,000.

Crisostomo L., Soriano A.M., Frost J.R., Olanubi O., Mendez M, & Pelka P. (2017). The Influence of E1A C-Terminus on Adenovirus Replicative Cycle. Viruses, 9(12), 387.

+ Open protocol

+ Expand

Western Blot Analysis of Protein Lysates

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

Proteins were isolated using RIPA lysis buffer supplemented with protease and phosphatase inhibitors (Life Technologies). For Western blot analysis 5 μg of protein lysates were loaded per lane on NuPage 10% Bis-Tris gels (Life Technologies) in 2-(N-morpholino)ethanesulfonic acid (MES) running buffer (Life Technologies). Samples were denatured using 10% mercaptoethanol at 95°C for 10 minutes before loading. Samples were transferred to Immobilon FL PVDF membranes (Millipore) and blocked in Li-cor blocking buffer for 1 hour. Primary antibodies were incubated overnight at 4°C and secondary IRDye antibodies were incubated at room temperature for 1 hour (Licor). The following primary antibodies were used for Western blotting: Actin antibody (Abcam, ab3280), EGF Receptor (Cell Signaling, CS#4267), Phospho-Akt (Ser473) (Cell Signaling, CS#4060), Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (Cell Signaling, CS#4370), N-Cadherin (BD Biosciences, 610921), E-Cadherin (BD Biosciences, 610182) and Cytokeratin 14 (Abcam, ab15461). Near-infrared fluorescence visualization was measured using Odyssey CLx scanner (Li-Cor, Cambridge, UK).

Choudhary K.S., Rohatgi N., Halldorsson S., Briem E., Gudjonsson T., Gudmundsson S, & Rolfsson O. (2016). EGFR Signal-Network Reconstruction Demonstrates Metabolic Crosstalk in EMT. PLoS Computational Biology, 12(6), e1004924.

+ Open protocol

+ Expand

Investigating Naringenin's Effects on ER+ Breast Cancer

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

MCF-7 ER+ breast cancer cells (HTB-22) were purchased from ATCC. Dulbecco's Modified Eagle Medium was purchased from Gibco. Charcoal-stripped fetal bovine serum, naringenin and 4-OH-tamoxifen were from Sigma Aldrich. Antibodies for ERK1/2, p-ERK1/2, AKT, p-AKT, caspase 7, PARP and U0126 were obtained from Cell Signaling. Guava Via-Count Reagent was purchased from Millipore. Actin antibody was obtained from Abcam. Anti-ERα antibody (HC-20) was from Santa Cruz biotechnology. AlexaFluor 488 conjugated Goat anti-Rabbit secondary antibody was obtained from Jackson ImmunoResearch. Anti-mouse and anti-rabbit horseradish peroxidase conjugated secondary antibodies were purchased from Sigma Aldrich. The enhanced chemiluminescence (ECL) detection kit was from BioExpress.

Eanes L, & Patel Y.M. (2016). Inhibition of the MAPK pathway alone is insufficient to account for all of the cytotoxic effects of naringenin in MCF-7 breast cancer cells. Biochimie Open, 3, 64-71.

+ Open protocol

+ Expand

Comparative Analysis of Cancer and Normal Cell Lines

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

The human malignant growth cancer cell lines MDA-MB-231, MCF-7, Skbr3 (breast cancer), HeLa (cervical cancer), HT-29, HCT 116 (colorectal adenocarcinoma) and normal cell line chang liver are procured from the National Centre for Cell Science, Pune, India. The cells were developed in Dulbecco’s Modified Eagle Medium (DMEM) medium enhanced with 10% inactivated Fetal Bovine Serum (FBS), 100 μg of streptomycin/ml, 100 U/ml of penicillin and incubated at 37 ± 2°C with 5% CO₂. MTT dye (3-(4, 5-dimethylthiazol-2-yl) - 2, 5-diphenyl tetrazolium bromide) was acquired from Sigma Aldrich (Mumbai, India). All the experimental reagents and solvents were purchased from sigma Aldrich chemicals Pvt Ltd. Recombinant VEGF₁₆₅ (Cell signalling technology), P-p44/42 MAPK, P-SAPK/JNK (G9) and JNK antibodies were purchased from cell signaling technology, USA. HRP-conjugated mouse secondary antibody and HRP-conjugated goat anti-rabbit secondary antibodies were purchased from cell signalling technology. Actin antibody was obtained from Abcam, USA.

Guruswamy D.K., Balaji K.D., Dharmappa K.K, & Jayarama S. (2020). Novel 3-(3, 5-difluoro-4-hydroxyphenyl)-1-(naphthalen-2-yl) prop-2-en-1-one as a potent inhibitor of MAP-kinase in HeLa cell lines and anti-angiogenic activity is mediated by HIF-1α in EAC animal model. Oncotarget, 11(50), 4661-4676.

+ Open protocol

+ Expand

Quantification of DNase Enzymes and Complexes

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

DNases were quantified using Human DNase 1 ELISA Kit (Nordic BioSite AB), Human DNase 1L3 ELISA Kit (Abbexa Ltd), and Human DNase 2A ELISA Kit (MyBioSource Inc) as indicated by the manufacturer’s instructions.
DNase 1L1 protein was quantified by flow cytometry using a rabbit anti-DNase 1L1 antibody (10 µg/mL, Atlas Antibodies), which was detected by an anti-rabbit Alexa647-conjugated secondary antibody (5 µg/mL, Biolegend).
For the detection of TREX 1 (three prime repair exonuclease 1) as well as DNase 1:actin complexes, custom-made ELISAs were used using 2 different TREX 1 antibodies (0.2 µg/mL, ThermoFisher; 0.6 µg/mL, horseradish peroxidase conjugated, Santa Cruz) or a DNase 1 (0.2 µg/mL, ThermoFisher) and an actin antibody (0.6 µg/mL, Abcam) as published previously.^{30 (link)} Details are provided in the Methods in the Data Supplement.

Haider P., Kral-Pointner J.B., Mayer J., Richter M., Kaun C., Brostjan C., Eilenberg W., Fischer M.B., Speidl W.S., Hengstenberg C., Huber K., Wojta J, & Hohensinner P. (2020). Neutrophil Extracellular Trap Degradation by Differently Polarized Macrophage Subsets. Arteriosclerosis, Thrombosis, and Vascular Biology, 40(9), 2265-2278.

+ Open protocol

+ Expand

Mitochondrial OXPHOS Protein Analysis

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

Mitochondria from the hippocampus of Crt^−/y and Crt^+/y mice were isolated as described above and separated on 12% Mini-PROTEAN® TGX pre-cast gels. Proteins were transferred to PVDF membranes and blocked using Odyssey blocking buffer (Li-Cor, Lincoln, Nebraska). Membranes were incubated with a Total OXPHOS Rodent WB Antibody Cocktail (Abcam, Cambridge, MA) at a 1:1000 dilution for 18 h at 4° C. This cocktail contains 5 antibodies, each directed at a specific protein in the OXPHOS complex. Complex 1 is probed using NDUF88, Complex II is represented by SDHB, Complex III by UQCRC2, Complex IV by MTCO1, and Complex V by ATP5A. The blot was concurrently probed with an Actin antibody (Abcam) at a concentration of 1:500 as a loading control. Following washing of the primary antibody, the blot was incubated with 1:10,000 dilution of IRDye 680RD Donkey anti-mouse secondary antibody (Li-Cor, Lincoln Nebraska) for the OXPHOS proteins and a IRDye 800CW Donkey anti-goat antibody for actin (Li-Cor). Fluorescent images were acquired on an Odyssey CLx (Li-Cor) imager and fluorescence was quantified using the software from the manufacturer. OXPHOS signals were normalized to the actin signal.

Perna M.K., Kokenge A.N., Miles K.N., Udobi K.C., Clark J.F., Pyne-Geithman G.J., Khuchua Z, & Skelton M.R. (2016). Creatine transporter deficiency leads to increased whole-body and cellular metabolism. Amino acids, 48(8), 2057-2065.

+ 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!