Mouse anti actin

Manufactured by Santa Cruz Biotechnology

Sourced in United States, United Kingdom

Mouse anti-actin is a primary antibody that specifically binds to the actin protein, a highly conserved cytoskeletal protein found in all eukaryotic cells. It can be used in various immunological techniques, such as Western blotting, immunohistochemistry, and immunocytochemistry, to detect and study the expression and localization of actin in biological samples.

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Lab products found in correlation

Pvdf membrane, by Merck Group (7 mentions) Protease inhibitor cocktail, by Merck Group (5 mentions) Supersignal west femto chemiluminescence kit, by Thermo Fisher Scientific (4 mentions) Rabbit anti scf, by Abcam (4 mentions) Pvdf membrane, by GE Healthcare (4 mentions) Bis tris polyacrylamide gel, by Thermo Fisher Scientific (4 mentions) Mouse anti gapdh, by Santa Cruz Biotechnology (3 mentions) Bradford assay, by Bio-Rad (3 mentions) Enhanced chemoluminescence, by PerkinElmer (3 mentions) Rabbit anti akt, by Cell Signaling Technology (3 mentions) Pvdf membrane, by Bio-Rad (3 mentions) Mouse anti smn, by BD (2 mentions) Phosstop, by Roche (2 mentions) Odyssey infrared imaging system, by LI COR (2 mentions) Rabbit anti atf4, by Cell Signaling Technology (2 mentions) Immunocruz western blotting luminol reagent, by Santa Cruz Biotechnology (2 mentions) Rabbit anti sam68, by Santa Cruz Biotechnology (2 mentions) Protease and phosphatase inhibitor, by Merck Group (2 mentions) Hrp conjugated goat anti mouse igg, by Southern Biotech (2 mentions) Mouse anti gapdh, by Abcam (2 mentions)

Close spelling variants of this product

Anti-actin (514 citations) Mouse anti-β-actin (325 citations) Mouse anti-β-actin monoclonal antibody (86 citations) Mouse monoclonal anti-β-actin (67 citations) Mouse anti-β-actin mAb (12 citations) Mouse anti-human β-actin monoclonal antibody (11 citations) Mouse monoclonal anti-β-actin (sc-47778) (6 citations) Mouse anti-human β-actin antibody (5 citations) Mouse anti-human β-ACTIN (sc-47778, (5 citations) Mouse anti-β-actin primary antibody (3 citations)

75 protocols using mouse anti actin

Protein Quantification and Western Blot Analysis

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The D17 cell line was treated with 20 µM GANT61 or vehicle DMSO for 96 hrs and cell lysate was made in lysis buffer. Protein was quantified using a BCA protein Assay Reagent (bicinchonic acid) (Pierce). A total of 40 µg of protein was loaded per sample in the 7.5% polyacrylamide gels under denaturing and reducing conditions and protein was transferred to nitrocellulose membranes. After transfer of protein, the membrane was probed and incubated overnight at 4°C with antibodies Rabbit anti-GLI1 (Abcam ab49314), Rabbit anti-GLI2 (Abcam ab26056), Rabbit anti-Patch1(Sigma-Aldrich P0088), Rabbit anti-Pax6 (Abcam ab5790), and Mouse anti-Actin (Santa Cruz sc-56459) in 5% non-fat milk. Then membranes were washed and subsequently exposed to the appropriate HRP-conjugated secondary antibodies for 1 hr. at room temperature. Bands were visualized by the enhanced chemiluminescence (Pierce) in Fluorchem E Imaging system (Protein Simple, CA). Constitutive expression levels of GLI1 and GLI2 were performed similarly, in the absence of treatment for Abrams, D17 and Moresco cell lines.

Shahi M.H., Holt R, & Rebhun R.B. (2014). Blocking Signaling at the Level of GLI Regulates Downstream Gene Expression and Inhibits Proliferation of Canine Osteosarcoma Cells. PLoS ONE, 9(5), e96593.

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Investigating C9orf72 and TFEB Regulation

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C9orf72-Flag and C9orf72 shRNA constructs were described previously [27 (link)]. GFP-TFEB (38119, Shawn Ferguson, Yale University), pRK5-HA GST RagB (19301, David Sabatini, Whitehead Institute), pRK5-HA GST RagC (19304, David Sabatini, Whitehead Institute), myc-Raptor (1859, David Sabatini, Whitehead Institute), pRK5-HA GST RagB 99L (19303, David Sabatini, Whitehead Institute), and pRK5-HA GST RagC 75N (19305, David Sabatini, Whitehead Institute) were obtained from Addgene.
Antibodies used in the human cell studies include the following: mouse anti-Flag (Sigma), mouse anti-GFP, rabbit anti-GAPDH (ThermoFisher), mouse anti-actin (Santa Cruz), mouse anti-C9orf72 (Bio-Rad), mouse anti-C9orf72 (Proteintech), mouse anti-HA (Bethyl Lab), rabbit anti p-70S6K, rabbit anti p-p70S6K, rabbit anti-RagA, rabbit anti-RagC, rabbit anti-Raptor, rabbit anti-LAMP1, rabbit anti-DYKDDDDK, rabbit anti-pTFEB, rabbit anti-TFEB (Cell Signaling), mouse anti-TFEB (Mybiosources), and rabbit anti-GFP (Abcam).

Ji Y.J., Ugolino J., Zhang T., Lu J., Kim D, & Wang J. (2020). C9orf72/ALFA-1 controls TFEB/HLH-30-dependent metabolism through dynamic regulation of Rag GTPases. PLoS Genetics, 16(4), e1008738.

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Bone Marrow Protein Extraction and Western Blot

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Bone marrow was flushed out of the bone and then dissociated in 66% Trichoracetic acid (TCA) in water. Extracts were incubated on ice for at least 15 min and centrifuged at 16,100 × g at 4°C for 10 min. Precipitates were washed in acetone twice and the dried pellets were solubilized in 9M urea, 2% TritonX-100, and 1% DTT. Samples were separated on 4–12% Bis-Tris polyacrylamide gels (Invitrogen) and transferred to PVDF membrane (Millipore). The blots were incubated with primary antibodies overnight at 4°C and then with secondary antibodies. Blots were developed with the SuperSignal West Femtochemiluminescence kit (Thermo Scientific). Primary antibodies used: rabbit-anti-SCF (Abcam, catalogue number ab64677, 1:1000) and mouse-anti-Actin (Santa Cruz, clone AC-15, 1:20,000).

Zhou B.O., Yu H., Yue R., Zhao Z., Rios J.J., Naveiras O, & Morrison S.J. (2017). Bone marrow adipocytes promote the regeneration of stem cells and hematopoiesis by secreting SCF. Nature cell biology, 19(8), 891-903.

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Western Blot Immunodetection Protocol

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Whole cell lysates were prepared by boiling cell pellets for 10 min in SDS lysis buffer [2% SDS, 10% Glycerol, 62 mM Tris-HCl, pH 6.8 and a complete mini-protease inhibitor cocktail (Roche Applied Science, Indianapolis, IN)]. After protein quantification with Bio-Rad Dc Protein Assay (Bio-Rad Laboratories, Hercules, CA), equal amounts of proteins were loaded, separated on a polyacrylamide gel, and transferred to a nitrocellulose membrane. Protein bands were immuno-detected with appropriate antibodies, e.g., rabbit-anti-XPC [47 (link)], mouse-anti-E-Cadherin (BD Transduction Laboratories, San Jose, CA, Cat. No. 610181, 1:1000), rabbit-anti-Snail (Cell Signaling, Danvers, MA, Cat. No. 3879, 1:1000), rabbit-anti-pERK1/2 (Cell Signaling, Cat. No. 9101, 1:1000), rabbit-anti-ERK2 (Cell Signaling, Cat. No. 9108, 1:1000), mouse-anti-γH2AX (Millipore, Billerica, MA, Cat. No. 05-636, 1:1000), rabbit-anti-H2AX (Cell Signaling, Cat. No. 7631, 1:1000), mouse-anti-Actin (Santa Cruz Biotechnology, Dallas, TX, Cat. No. sc-47778, 1:1000), and goat-anti-Lamin B (Santa Cruz Biotechnology, Cat. No. sc-6216, 1:1000).

Cui T., Srivastava A.K., Han C., Yang L., Zhao R., Zou N., Qu M., Duan W., Zhang X, & Wang Q.E. (2015). XPC inhibits NSCLC cell proliferation and migration by enhancing E-Cadherin expression. Oncotarget, 6(12), 10060-10072.

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Evaluating Inflammatory Markers in Cells

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Recombinant human TNF-α was purchased from Pepro Tech. Blocking mAb against β2 integrin (TS1/18) and MHC class I mAb (W6/32) were purchased from American Type Culture Collection (ATCC). The following primary antibodies were used: rat anti-human EMCN (1:1,000, ab45771, Abcam), rat anti-mouse EMCN V.7C7 (1:1,000, sc-65495, Santa Cruz Biotechnology), mouse anti-actin (1:1,000, Santa Cruz Biotechnology), mouse anti-tubulin (1:2,000, CalBiochem) and rabbit anti-GAPDH (1:1,000, Santa Cruz Biotechnology). Antisera against human E-selectin (clone CL2), VCAM-1 (clone E1/6) and ICAM-1 (clone Hu 5/3) were purchased from ATCC. CD31 was detected using rat anti-mouse CD31 (1:250, BD Pharmingen). For flow cytometry of cells from ciliary body, PE-NIMP-R14 (Novus), Brilliant Violet 421 anti-CD45 and PE-Cy7 anti-F4/80 (Biolegend) and PE-CD31 (BD Pharmingen) or their corresponding isotype controls were used.

Zahr A., Alcaide P., Yang J., Jones A., Gregory M., dela Paz N.G., Patel-Hett S., Nevers T., Koirala A., Luscinskas F.W., Saint-Geniez M., Ksander B., D'Amore P.A, & Argüeso P. (2016). Endomucin prevents leukocyte–endothelial cell adhesion and has a critical role under resting and inflammatory conditions. Nature Communications, 7, 10363.

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Splenic Endothelial Cell Protein Extraction and Analysis

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Approximately 30,000 CD45⁻Ter119⁻VE-cadherin⁺ splenic endothelial cells were flow cytometrically sorted into 50 μl of 66% Trichoracetic acid (TCA) in water. Extracts were incubated on ice for at least 15 min and centrifuged at 16,100 × g at 4°C for 10 min. Precipitates were washed in acetone twice and the dried pellets were solubilized in 9M urea, 2% TritonX-100, and 1% DTT. Samples were separated on 4-12% Bis-Tris polyacrylamide gels (Invitrogen) and transferred to PVDF membrane (Millipore). The blots were incubated with primary antibodies overnight at 4°C and then with secondary antibodies. Blots were developed with the SuperSignal West Femtochemiluminescence kit (Thermo Scientific). Primary antibodies used: rabbit-anti-SCF (Abcam, 1:1000) and mouse-anti-Actin (Santa Cruz, clone AC-15, 1:20,000).

Inra C.N., Zhou B.O., Acar M., Murphy M.M., Richardson J., Zhao Z, & Morrison S.J. (2015). A perisinusoidal niche for extramedullary haematopoiesis in the spleen. Nature, 527(7579), 466-471.

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Quantifying Protein Expression with Western Blot

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Western blot assays were performed using the following primary antibodies: goat anti-human legumain (AF2199; R&D); mouse anti-actin (Santa Cruz Biotechnology); rabbit anti-human MMP2 (1:500; Abcam); anti-MMP9 (1:1,000; Cell Signaling). Briefly, stimulated cells were lysed with RIPA buffer [50 mM Tris–HCl (pH 7.5), 150 mM NaCl, 1% Triton X-100] containing protease inhibitors (Complete Mini; Roche); 25-µg samples of the lysates were separated on 8–10% SDS-PAGE gels and transferred to PVDF membranes. The membranes were incubated with primary antibodies overnight at 4°C. The primary antibody incubation was followed by incubation with an HRP-conjugated secondary antibody. The bound antibodies were detected using an ECL kit (PI32209; Pierce).

Meng F, & Liu W. (2016). Knockdown of Legumain Suppresses Cervical Cancer Cell Migration and Invasion. Oncology Research, 23(1-2), 7-12.

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Antibodies and Plasmid Constructs for FAF1 Study

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The following antibodies were used in this study: mouse monoclonal Flag antibody (Sigma), rabbit anti-FAF1 (AbFrontier), rabbit anti-NMIIA antibody (BioLegend), mouse anti-RhoA (Abcam), mouse anti-p190B RhoGAP, mouse anti-β-catenin, mouse anti-γ-catenin, mouse-anti-N-cadherin (BD Bioscience), rabbit anti-MLC2, anti-PP-MLC2 (Cell Signaling Technology), rabbit anti-IQGAP1, mouse anti-actin, and anti-tubulin (Santa Cruz Biotechnology). Y-27632 (Y0503) was purchased from Sigma. GST-FAF1 WT, GST-FAF1 UAS-UBX, pFlag-CMV-2-FAF1 WT, pFlag-CMV-2-FAF1 (82–650), and pFlag-CMV-2-FAF1 (1–120) were prepared as previously described (Murshudov et al., 1997 (link); Song et al., 2005 (link)). pFlag-CMV-2-FAF1 H160A was generated by cloning employing mutagenesis. All plasmid constructs were verified by DNA sequencing.

Song S., Park J.K., Shin S.C., Lee J.J., Hong S.K., Song I.K., Kim B., Song E.J., Lee K.J, & Kim E.E. (2022). The complex of Fas-associated factor 1 with Hsp70 stabilizes the adherens junction integrity by suppressing RhoA activation. Journal of Molecular Cell Biology, 14(6), mjac037.

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Western Blot Analysis of Protein Targets

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Western blot analysis was performed as previously described (Paronetto et al., 2007 (link)). The primary antibodies (1:1,000) were rabbit anti-SAM68, mouse anti-ACTIN, mouse anti-GAPDH (all from Santa Cruz Biotechnology, Inc.), mouse anti-U2AF65 (Sigma-Aldrich), and mouse anti-SMN (BD).

Pagliarini V., Pelosi L., Bustamante M.B., Nobili A., Berardinelli M.G., D’Amelio M., Musarò A, & Sette C. (2015). SAM68 is a physiological regulator of SMN2 splicing in spinal muscular atrophy. The Journal of Cell Biology, 211(1), 77-90.

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Adipose JNK Activity Immunoblotting

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For adipose JNK activity, WAT from LysM/K2KO CD/HFD-fed mice was lysed and equal amounts of protein were immunoblotted after SDS-PAGE resolution. Antibodies used for immunoblotting are as follows: rabbit anti-p-JNK (1:2000, EMD Millipore, 07-175), rabbit anti-SAPK/JNK (1:2000, Cell Signaling, 9252 S), mouse anti-actin (1:5000, Santa Cruz, sc-47778).

Sweet D.R., Vasudevan N.T., Fan L., Booth C.E., Keerthy K.S., Liao X., Vinayachandran V., Takami Y., Tugal D., Sharma N., Chan E.R., Zhang L., Qing Y., Gerson S.L., Fu C., Wynshaw-Boris A., Sangwung P., Nayak L., Holvoet P., Matoba K., Lu Y., Zhou G, & Jain M.K. (2020). Myeloid Krüppel-like factor 2 is a critical regulator of metabolic inflammation. Nature Communications, 11, 5872.

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