Sc 1751

Manufactured by Santa Cruz Biotechnology

Sourced in United Kingdom

Sc-1751 is a laboratory equipment product manufactured by Santa Cruz Biotechnology. It is a high-quality, versatile instrument designed for use in various scientific research applications. The core function of Sc-1751 is to assist in the analysis and processing of biological samples.

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Lab products found in correlation

Lsm 510, by Zeiss (2 mentions) Lsm 780, by Zeiss (2 mentions) Ab20841, by Abcam (1 mentions) Ab12148, by Abcam (1 mentions) Sc 32318, by Santa Cruz Biotechnology (1 mentions) Myc tag 9b11, by Cell Signaling Technology (1 mentions) Alexa fluor conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions) Axioplan 2 microscope, by Zeiss (1 mentions) Blotto non fat dry milk, by Santa Cruz Biotechnology (1 mentions) Ab34710, by Abcam (1 mentions) Bca assay, by Thermo Fisher Scientific (1 mentions) Sc 8035, by Santa Cruz Biotechnology (1 mentions) Sc 47724, by Santa Cruz Biotechnology (1 mentions) Image lab software, by Bio-Rad (1 mentions) Ab5694, by Abcam (1 mentions) Ab5392, by Abcam (1 mentions) A21207, by Thermo Fisher Scientific (1 mentions) Mab2160, by Merck Group (1 mentions) Alexa fluor 488 conjugated donkey anti rabbit, by Thermo Fisher Scientific (1 mentions) A21203, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Goat polyclonal anti-TIA-1 (sc-1751) (2 citations)

8 protocols using sc 1751

Immunostaining and Stress Granule Analysis

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Cells grown on glass coverslips were fixed and immunostained according to the protocol in [6] (link) using mouse monoclonal antibodies to IAV nucleoprotein (AAH5; Abcam), G3BP (clone 23, BD Transduction Labs), and PABP1 (sc-32318, Santa Cruz Biotechnology); goat polyclonal antibody to TIA-1 (sc-1751, Santa Cruz Biotechnology), influenza virus (ab20841, Abcam), rabbit antibody to TIAR (clone D32D3, Cell Signaling), YB-1 (ab12148, Abcam) and myc-tag (9B11; Cell Signaling) at manufacturer-recommended dilutions. AlexaFluor-conjugated secondary antibodies (Molecular Probes) were used at 1∶1000 dilution. Images were captured using Zeiss Axioplan II microscope or Zeiss LSM 510 laser scanning microscope. Quantification of stress granules was done in at least 3 random fields of view with greater than 200 cells analysed on each slide. Cells were considered stress granule-positive if two or more stress granule marker foci were present in the cytoplasm. For western blot analysis, whole cell lysates were resolved on denaturing 10% polyacrylamide gels and analyzed using primary antibodies described above and the antibodies to phospho-Ser-51- eIF2α (rabbit, D9G8, Cell Signaling), eIF4A (goat, sc-14211, Santa Cruz Biotechnology), and actin (rabbit, 4968; Cell Signaling).

Khaperskyy D.A., Emara M.M., Johnston B.P., Anderson P., Hatchette T.F, & McCormick C. (2014). Influenza A Virus Host Shutoff Disables Antiviral Stress-Induced Translation Arrest. PLoS Pathogens, 10(7), e1004217.

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Western Blot Analysis of Protein Expression

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After 24 h IL-1β stimulation, F-NL cells were lysed with RIPA buffer and frozen at −80 °C. Proteins were purified with high speed centrifugation and their concentration was determined by bicinchoninic acid (BCA) assay (ThermoFisher, Waltham, MA, USA). Proteins were diluted with 4× Laemmli buffer and boiled for 10 min. 10–20 μg of total protein were separated with SDS-PAGE and transferred into PVDF membrane. After 1 h blocking with 5% Blotto non-fat dry milk (Santa Cruz, Active Santa Cruz, CA, USA) in TBS 0.05% tween 20, the membranes were incubated with specific antibodies recognizing human forms of COX-2 (160112, Cayman Chemical), GAPDH (sc-47724, Santa Cruz), α-Tubulin (sc-8035, Santa Cruz), α-SMA (ab5694, Abcam, Cambridge, UK), COL1 (ab34710, Abcam) and TIA-1 (sc-1751, Santa Cruz). The optical densitometry (OD) of the protein bands was analyzed using Image Lab™ Software (Bio-Rad, Hercules, CA, USA). Data were normalized with the loading control GAPDH or α-Tubulin and fold changes from control condition were calculated.

Pasini A., Brand O.J., Jenkins G., Knox A.J, & Pang L. (2018). Suberanilohydroxamic acid prevents TGF-β1-induced COX-2 repression in human lung fibroblasts post-transcriptionally by TIA-1 downregulation. Biochimica et Biophysica Acta, 1861(5), 463-472.

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Comprehensive Immunostaining Protocol for Protein Detection

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The following primary antibodies were used: anti-GFP (1:500; 632380, Clontech), anti-HuR (1:200; 11910-1-AP, Proteintech), anti-DNAPTP6 (1:300; 16938-1-AP, Proteintech), anti-MAP2 (1:1000; ab5392, Abcam), anti-G3BP1 (1:300; 13057-2-AP, Proteintech), recombinant BG4 (anti-G4, manually prepared, used at 1 μg/ml), anti-6×His (1:1000; ab18184, Abcam), anti-TIA1 (1:1000; sc-1751, Santa Cruz Biotechnology), anti-Tuj1 (tubulin β-3) (1:1000; 802001, BioLegend), anti–cleaved caspase-3 (1:200; ab2302, Abcam), anti-DYKDDDDK tag (1:800; 14793S, Cell Signaling Technology), anti-FLAG (1:1000; F1804, Sigma-Aldrich), anti-FMRP (1:1000; MAB2160, Millipore), and anti-digoxigenin (1:1000; ab76907, Abcam). For immunocytochemistry and immunohistochemistry, the following secondary antibodies were used: Alexa Fluor 488–conjugated donkey anti-rabbit (1:500; A-21206, Thermo Fisher Scientific), Alexa Fluor 594–conjugated donkey anti-rabbit (1:500; A-21207, Thermo Fisher Scientific), Alexa Fluor 488–conjugated donkey anti-mouse (1:500; A-21202, Thermo Fisher Scientific), Alexa Fluor 594–conjugated donkey anti-mouse (1:500; A-21203, Thermo Fisher Scientific), Alexa Fluor 594–conjugated donkey anti-goat (1:500; A-11058, Thermo Fisher Scientific), and DyLight 405–AffiniPure donkey anti-chicken IgY (1:500; 703-475-155, Jackson ImmunoResearch).

Asamitsu S., Yabuki Y., Matsuo K., Kawasaki M., Hirose Y., Kashiwazaki G., Chandran A., Bando T., Wang D.O., Sugiyama H, & Shioda N. (2023). RNA G-quadruplex organizes stress granule assembly through DNAPTP6 in neurons. Science Advances, 9(8), eade2035.

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Visualizing FUS Protein Localization

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N2A cells or primary cortical neurons were seeded on gelatin or poly-d-lysine hydrobromide-coated glass coverslips. Twenty-four hours after the transfection with EGFP-FUS, cells were rinsed with 1× PBS, fixed with 4% formaldehyde in 1× PBS, and permeabilized with 0.25% Triton X-100 in 1× PBS. Primary fibroblast cells were cultured, fixed, and permeabilized similarly as above. The primary antibodies used in this study were mouse anti-FUS (sc-47711; Santa Cruz Biotechnology, Santa Cruz, CA), rabbit anti-G3BP1 (13057-2-AP; Proteintech), and goat anti-TIA1 (sc-1751; Santa Cruz). The secondary antibodies were Alexa Fluor 488 donkey anti-mouse (A-21202; Life Technologies), Alexa Fluor 647 donkey anti-rabbit (A-31573; Life Technologies), and Alexa Fluor 568 donkey anti-goat (A-11057; Life Technologies). The samples were mounted using Vectashield Mounting Medium (Vector Laboratories, Burlingame, CA). Images were acquired using a Nikon A1 confocal microscope with a 60× objective.

Kuang L., Kamelgarn M., Arenas A., Gal J., Taylor D., Gong W., Brown M., St. Clair D., Kasarskis E.J, & Zhu H. (2017). Clinical and experimental studies of a novel P525R FUS mutation in amyotrophic lateral sclerosis. Neurology: Genetics, 3(4), e172.

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Western Blot Analysis of Protein Markers

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Cells were washed twice with sterile PBS and then lysed on ice for 10 min in lysis buffer [17 (link)]. Lysates were centrifuged at 13,000 rpm for 10 min at 4 °C. Supernatants were collected and protein concentration was determined by the Bradford method. Samples were stored at − 80 °C until used. Proteins were separated on a 10% SDS polyacrylamide gel (10 μg of total proteins per well) and transferred to a polyvinylidene difluoride membrane. Membranes were incubated for 1 h in TBST 5% dried milk to saturate all non-specific binding sites. Incubation with primary antibodies was overnight at 4 °C, using mouse anti-DJ-1 antibody (1:1000; sc-55572, Santa Cruz Biotechnology), rabbit anti-β-tubulin (1:1000; #2128, Cell Signaling Technology), rabbit anti-eIF4A3 (1:1000; ab32485, Abcam), or goat anti-TIA1 (1:200; sc-1751, Santa Cruz Biotechnology). Blots were developed using horseradish peroxidase (HRP)-conjugated secondary antibodies (1:10000; Vector Laboratories) and the ECL chemiluminescence system (SuperSignal West Dura Extended Duration Substrate, Thermo Scientific).

Repici M., Hassanjani M., Maddison D.C., Garção P., Cimini S., Patel B., Szegö É.M., Straatman K.R., Lilley K.S., Borsello T., Outeiro T.F., Panman L, & Giorgini F. (2018). The Parkinson’s Disease-Linked Protein DJ-1 Associates with Cytoplasmic mRNP Granules During Stress and Neurodegeneration. Molecular Neurobiology, 56(1), 61-77.

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Immunostaining of Stress Granules

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Cells grown on glass coverslips were fixed using 4% paraformaldehyde in PBS for 10 min at room temperature and permeabilized with PBS containing 0.1% Triton X-100 at room temperature for 10 min. After blocking with 1% bovine serum albumin in PBST (PBS + 0.1% Tween 20) for 30 min, SGs were immunostained with rabbit anti-PABPC1 (Abcam, ab21060, 1:500 in PBST) or goat anti-TIA1 (Santa Cruz Biotechnology, sc-1751, 1:500 in PBST) for 60 min. FITC-conjugated goat anti-rabbit antibody (Jackson ImmunoResearch, 111-095-144, 1:500 in PBST) or Texas red-conjugated donkey anti-goat (Jackson ImmunoResearch, 705-075-147, 1:500 in PBST) was applied at RT for 60 min. Cells were mounted on glass slides in SlowFade Gold Antifade reagent (Thermo Fisher) with DAPI. Fluorescence images were collected using the EVOS FL Auto Cell Imaging System (Thermo Fisher).

Zheng D., Wang R., Ding Q., Wang T., Xie B., Wei L., Zhong Z, & Tian B. (2018). Cellular stress alters 3′UTR landscape through alternative polyadenylation and isoform-specific degradation. Nature Communications, 9, 2268.

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Immunofluorescence Assay for SG Markers

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Cells were fixed for 20 min in 4% formaldehyde, 1X PBS, followed by permeabilization for 10 min with 0.5% Triton, 1X PBS. Cells were rinsed with 1X PBS and blocked with blocking buffer (1X PBS, 2% BSA, 0.02% Triton). Cells were incubated with the primary antibodies against SG marker like TIA1 (TIA1, dilution 1:100, cat.# sc-1751,Santacruz) and antibodies against RBPs Sundararaman et al., 2016 (link)) diluted in blocking buffer for 2 hour at room temperature or overnight at 4°C. Then, the cells were thoroughly washed with 1X PBS, 0.2% Tween 20, and incubated for 2 hour with secondary antibodies (Alexa Fluor 647, cat. # A21447, Alexa Fluor 594, cat. # A21207, Life technologies and Alexa Fluor 488, cat. # 111-546-144, JacksonImmuno, dilution 1:500) diluted in blocking buffer. Cells were washed, incubated for 5 min with DAPI and washed again. Cells were stored in the dark at 4°C in 1X PBS or 50% glycerol/PBS for long-term storage. All images were taken using high content screen microscopy, ImageXpress Micro.

Markmiller S., Soltanieh S., Server K.L., Mak R., Jin W., Fang M.Y., Luo E.C., Krach F., Yang D., Sen A., Fulzele A., Wozniak J.M., Gonzalez D.J., Kankel M.W., Gao F.B., Bennett E.J., Lécuyer E, & Yeo G.W. (2018). Context-Dependent and Disease-Specific Diversity in Protein Interactions within Stress Granules. Cell, 172(3), 590-604.e13.

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Immunostaining of Oocytes

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Immunostaining of oocytes was done as described (Gagnon and Mowry, 2011) . The primary anti-Tia1 antibody (sc-1751, Santa Cruz) was diluted 1:250 and the secondary anti-goat-Alexa633 antibody (A21082, Invitrogen) was diluted 1:500 in PBT + 2 % horse serum and 2 % BSA. Fluorescence was visualized by confocal imaging (LSM780, Zeiss).

Bauermeister D., Claußen M, & Pieler T. (2015). T-cell internal antigen 1 counteracts somatic RNA degradation during early Xenopus embryogenesis. The International journal of developmental biology, 59(10-12).

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