Log In Sign up
The largest database of trusted experimental protocols

Adi spa 901

Manufactured by Enzo Life Sciences
Visit Supplier
Sourced in United States

The ADI-SPA-901 is a high-performance automated dispenser designed for precise and consistent liquid handling. It is capable of aspirating and dispensing a wide range of sample volumes with a high degree of accuracy and repeatability. The device is suitable for use in various laboratory applications that require accurate liquid transfer.

Automatically generated - may contain errors

Lab products found in correlation

Ideal chip seq kit for transcription factors, by Diagenode (3 mentions) Ab76076, by Abcam (3 mentions) Adi spa 810, by Enzo Life Sciences (3 mentions) Lsm 710 confocal microscope, by Zeiss (3 mentions) T8328, by Merck Group (2 mentions) Elisa, by R&D Systems (2 mentions) Nextflex chip seq kit and barcodes, by PerkinElmer (2 mentions) Normal rabbit igg, by Merck Group (2 mentions) Bioruptor pico, by Diagenode (2 mentions) T5168, by Merck Group (2 mentions) Ecl kit, by Thermo Fisher Scientific (2 mentions) Ab77231, by Abcam (1 mentions) Sc 899, by Santa Cruz Biotechnology (1 mentions) Ab1791, by Abcam (1 mentions) Sc 5286, by Santa Cruz Biotechnology (1 mentions) Mms 126r, by Fortrea (1 mentions) Streptavidin donor bead, by PerkinElmer (1 mentions) Nuclear extraction kit, by Active Motif (1 mentions) Optiplate, by PerkinElmer (1 mentions) Envision multilabel plate reader, by PerkinElmer (1 mentions)

18 protocols using adi spa 901

1

ChIP-seq of Heat Shock Factor 1

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
ChIP-seq of HSF1 was performed according to the method described by [17 (link)]. The HSF1 polyclonal antibody was from Stressgen (Enzo Life Sciences, ADI-SPA-901). For each IP, 5 μl of HSF1 antibody was used with 250 μl of precleared chromatin. A ChIP library was prepared using four independent ChIP experiments at ZT14, and one lane was sequenced to obtain about 20 million uniquely mapped reads.
Sobel J.A., Krier I., Andersin T., Raghav S., Canella D., Gilardi F., Kalantzi A.S., Rey G., Weger B., Gachon F., Dal Peraro M., Hernandez N., Schibler U., Deplancke B, & Naef F. (2017). Transcriptional regulatory logic of the diurnal cycle in the mouse liver. PLoS Biology, 15(4), e2001069.
+ Open protocol
+ Expand
2

Antibody-based Immunoblotting and ChIP-seq

Check if the same lab product or an alternative is used in the 5 most similar protocols
Antibodies used for immunoblotting were anti-SUMO2/3 (M114-3, MBL), anti-UBC9 (sc10759, Santa Cruz Biotechnology), anti-PIAS1 (ab77231, Abcam), anti-tubulin (sc5286, Santa Cruz Biotechnology), anti-laminB1 (sc6216, Santa Cruz Biotechnology), anti-Pol2 (sc-899, Santa Cruz Biotechnology), anti-histone H3 (ab1791, Abcam), and anti-HSF1 (ADI-SPA-901; ENZO Life Sciences). Antibodies used for ChIP-seq were anti-SUMO2/3 (M114-3, MBL), anti-PIAS1 (ab77231, Abcam), and anti-Pol2 (MMS-126R, Covance).
Niskanen E.A., Malinen M., Sutinen P., Toropainen S., Paakinaho V., Vihervaara A., Joutsen J., Kaikkonen M.U., Sistonen L, & Palvimo J.J. (2015). Global SUMOylation on active chromatin is an acute heat stress response restricting transcription. Genome Biology, 16(1), 153.
+ Open protocol
+ Expand
3

Quantifying HSF1 Binding to HSE

Check if the same lab product or an alternative is used in the 5 most similar protocols
Biotinylated and standard oligonucleotides were purchased from Integrated DNA Technologies (IDT), diluted in Tris-EDTA buffer (pH 8.0) and annealed. Cells were treated (heat shocked for 15 min at 43 °C (HS) or treated for 1 h with 500 ng/mL CD40 ligand, 100 ng/mL IL-4, or 100 ng/mL IL-21) and nuclear extracts were prepared using the Nuclear Extraction kit (Active Motif). For the entire assay, solutions were diluted to their working concentrations in AlphaLISA Immunoassay buffer solution 5X (AL001F). Protein A acceptor beads (6760137, PerkinElmer) were incubated with polyclonal rabbit HSF1 antibody (ADI-SPA-901, Enzo Life Sciences) for 1 h at room temperature with agitation. Streptavidin donor beads (6760002, PerkinElmer) were added and the mixture was kept in the dark with agitation. Meanwhile, 2 μg of nuclear extract were incubated with annealed biotinylated HSPA1A HSE or mutated HSE for 30 min in white 384-well Optiplates (6005620, PerkinElmer). A mixture of acceptor and donor beads was added to each well and incubated for 1 h. Luminescence was measured in an EnVision Multilabel Plate Reader (PerkinElmer).
Fernando T.M., Marullo R., Gresely B.P., Phillip J.M., Yang S.N., Lundell-Smith G., Torregroza I., Ahn H., Evans T., Győrffy B., Privé G.G., Hirano M., Melnick A.M, & Cerchietti L. (2019). BCL6 evolved to enable stress tolerance in vertebrates and is broadly required by cancer cells to adapt to stress. Cancer discovery, 9(5), 662-679.
+ Open protocol
+ Expand
4

Immunofluorescence Imaging of HSP90, HSF1, and ESR1

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were plated onto Nunc Lab-Tek II chambered coverglass (#155383, Nalge Nunc International, Rochester, NY) and fixed for 15 min with 4% PFA solution in PBS, washed, treated with 0.1% Triton-X100 in PBS for 5 min, and washed again in PBS (3 × 5 min). IF imaging was performed using primary antibodies: anti-HSP90 (1:200; ADI-SPA-836, Enzo Life Science), anti-HSF1 (1:300; ADI-SPA-901, Enzo Life Sciences), or anti-ESR1 (1:200; C15100066, Diagenode) and secondary Alexa Fluor (488 or 594) conjugated antibodies (Abcam). Finally, the DNA was stained with DAPI. Images were taken using Carl Zeiss LSM 710 confocal microscope with ZEN navigation software.
Vydra N., Janus P., Kus P., Stokowy T., Mrowiec K., Toma-Jonik A., Krzywon A., Cortez A.J., Wojtas B., Gielniewski B., Jaksik R., Kimmel M, & Widlak W. (2021). Heat shock factor 1 (HSF1) cooperates with estrogen receptor α (ERα) in the regulation of estrogen action in breast cancer cells. eLife, 10, e69843.
+ Open protocol
+ Expand
5

Quantitative Western Blot Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were lysed in Laemmli sample buffer (30% glycerol; 3% SDS; 188 mM Tris–Cl, pH 6.8; 0.015% bromophenol blue; 3% β-mercaptoethanol). Equal volumes of lysates were run on SDS-PAGE, after which proteins were transferred to nitrocellulose membrane. Membranes were blocked with nonfat dried milk diluted in PBS-Tween20 for 1 h at room temperature (RT). Proteins bound to membrane were analyzed using primary antibodies against HSF1 (ADI-SPA-901, Enzo), HSF2 (3E2, EMD Millipore) and β-tubulin (T8328, Merck). Next, the membranes were incubated in secondary HRP-conjugated antibodies, and the proteins were detected with enhanced chemiluminescence.
Himanen S.V., Puustinen M.C., Da Silva A.J., Vihervaara A, & Sistonen L. (2022). HSFs drive transcription of distinct genes and enhancers during oxidative stress and heat shock. Nucleic Acids Research, 50(11), 6102-6115.
+ Open protocol
+ Expand
6

ChIP-seq and ChIP-qPCR of HSF1 in Heat-Shocked MCF7 Cells

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
The ChIP assay in MCF7 wild-type cells, untreated and heat-shocked at 43 °C for 15 min, was performed according to the protocol from the iDeal ChIP-seq Kit for Transcription Factors (Diagenode, Denville, NJ, USA) using anti-HSF1 antibody (#ADI-SPA-901, Enzo), sequenced, and analyzed as described in detail in [26 (link)]. ChIP-Seq heatmaps were prepared using the peakHeatmap function from the ChIPseeker Bioconductor package (v. 1.26.2), with margins of 3000 nucleotides upstream and downstream from the promoter. The raw ChIP-seq data were deposited in the NCBI GEO database; acc. no. GSE137558 (GSM4081758, GSM4081759, and GSM4081762). For ChIP-qPCR (described in detail in [26 (link)]) in HSF1+ and HSF1− MCF7 cells, anti-HSF2 antibody (#AF5227, R&D Systems) was used in addition to anti-HSF1. The sequences of used primers are presented in Table S2.
Janus P., Kuś P., Vydra N., Toma-Jonik A., Stokowy T., Mrowiec K., Wojtaś B., Gielniewski B, & Widłak W. (2022). HSF1 Can Prevent Inflammation following Heat Shock by Inhibiting the Excessive Activation of the ATF3 and JUN&FOS Genes. Cells, 11(16), 2510.
+ Open protocol
+ Expand
7

Western Blot and ELISA Characterization

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were lysed in TBST buffer as described previously [30 (link)]. The protein content was determined using the BCA™ Protein Assay Kit (Pierce). After transfer on PVDF membranes (BioRad) blots were blocked in 5% skim milk and incubated with antibodies (4 °C, overnight) directed against HSF-1 (ADI-SPA-901; Enzo Life Sciences Inc., Farmingdale, NY, USA), HSF-1 phospho S326 (pHSF-1) (ab76076; abcam), Hsp70 (ADI-SPA-810; Enzo Life Sciences; cmHsp70.1; multimmune GmbH), Hsp27 (ADI-SPA-800; Enzo Life Sciences), Akt (Cell Signaling Technology Europe, Frankfurt, Germany) and β-actin (A5316; Sigma-Aldrich, St. Louis, MO, USA). Horseradish-peroxidase (HRP)-conjugated anti-rabbit/anti-mouse antibodies (Promega, Fitchburg, WI, USA, W401B/W402B) were used as secondary antibodies. Immune complexes as detected by ECL detection system (GE Healthcare, Chicago, IL, USA) were imaged digitally (ChemiDoc Touch Imaging System, Biorad, Hercules, CA, USA). Hsp70 protein concentrations in cell lysates were quantified by ELISA (R&D systems, Minneaplis, MN, USA) following the manufacturer’s recommendations.
Kühnel A., Schilling D., Combs S.E., Haller B., Schwab M, & Multhoff G. (2019). Radiosensitization of HSF-1 Knockdown Lung Cancer Cells by Low Concentrations of Hsp90 Inhibitor NVP-AUY922. Cells, 8(10), 1166.
+ Open protocol
+ Expand
8

Heat Stress Response Regulation

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were either exposed to 10 mM MβCD for 10 min [18 (link)] or to 50 µg/mL nystatin for 1 h [20 (link)] at 37 °C followed by heat stress for the indicated time at 42 °C in serum-supplemented RPMI medium. Immediately after heat stress, cells were lysed in RIPA buffer and HSF1 (ADI-SPA-901, Enzo Life Sciences; RT-405, Thermo-Scientific, Waltham, MA, USA) and GAPDH (G9545, Sigma-Aldrich) were analyzed through western blotting with the indicated antibodies. Signals were visualized by the use of HRP-conjugated secondary antibodies.
Crul T., Csoboz B., Gombos I., Marton A., Peter M., Balogh G., Vizler C., Szente L, & Vigh L. (2020). Modulation of Plasma Membrane Composition and Microdomain Organization Impairs Heat Shock Protein Expression in B16-F10 Mouse Melanoma Cells. Cells, 9(4), 951.
+ Open protocol
+ Expand
9

Mapping HSF1 and HSF2 Binding Sites

Check if the same lab product or an alternative is used in the 5 most similar protocols
HSF1- and HSF2-bound DNA fragments were isolated from two biological replicates using ChIP as previously described (26 (link)). Specifically, ChIP-seq was performed in WT MEFs that were untreated, exposed to 30 μM menadione for 2 h or heat-shocked at 42°C for 1 h. Cells were crosslinked with 1% paraformaldehyde for 5 min, after which paraformaldehyde was quenched with 125 mM glycine. Cells were lysed and the chromatin was fragmented by sonication with Bioruptor Pico (Diagenode) using seven cycles (30 s on/off). Agarose gel electrophoresis was used to verify that fragment size after sonication was 300–400 bp. The following antibodies were used for immunoprecipitation: HSF1 (ADI-SPA-901, Enzo), HSF2 (26 (link)), and normal rabbit IgG (EMD Millipore). Crosslinks were reversed by incubating the samples at 65°C overnight, and the DNA was purified with phenol:chloroform. ChIP-seq libraries were generated using NEXTFLEX ChIP-seq kit and barcodes (Perkin Elmer). NovaSeq 6000 was used to sequence ChIP-seq libraries. The raw files are available in GEO accession: GSE183245.
Himanen S.V., Puustinen M.C., Da Silva A.J., Vihervaara A, & Sistonen L. (2022). HSFs drive transcription of distinct genes and enhancers during oxidative stress and heat shock. Nucleic Acids Research, 50(11), 6102-6115.
+ Open protocol
+ Expand
10

Quantification of MICA and MICB

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were lysed in TBST buffer as described previously [18 (link)]. The protein content in the cell lysates was determined using the BCA™ Protein Assay Kit (Pierce). On immunoblots, proteins were detected with antibodies against HSF1 (ADI-SPA-901; Enzo Life Sciences), HSF1 phospho S326 (pHSF1) (ab76076; abcam), Hsp70 (ADI-SPA-810; Enzo Life Sciences) and β-actin (A5316; Sigma-Aldrich).
MICA and MICB concentrations in the cell lysates were measured by ELISA (R&D Systems), and the concentrations were calculated relative to the total protein content of each sample.
Schilling D., Kühnel A., Tetzlaff F., Konrad S, & Multhoff G. (2015). NZ28-induced inhibition of HSF1, SP1 and NF-κB triggers the loss of the natural killer cell-activating ligands MICA/B on human tumor cells. Cancer Immunology, Immunotherapy, 64(5), 599-608.
+ 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?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!