A21240

Manufactured by Thermo Fisher Scientific

Sourced in United States

The A21240 is a laboratory instrument designed for nucleic acid quantification. It utilizes a specialized detection method to determine the concentration of DNA or RNA samples. The device provides accurate and reliable measurements to support research and diagnostic applications.

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Lab products found in correlation

A11008, by Thermo Fisher Scientific (3 mentions) A11034, by Thermo Fisher Scientific (3 mentions) Axio observer z1, by Zeiss (3 mentions) Hoechst dye, by Thermo Fisher Scientific (2 mentions) Multispectral imaging system, by Nikon (2 mentions) A 21244, by Thermo Fisher Scientific (2 mentions) Lsm800 confocal microscope, by Zeiss (2 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (2 mentions) Lsm 880, by Zeiss (2 mentions) Axiovert 200m, by Zeiss (2 mentions) Diva buffer, by Biocare Medical (1 mentions) Mca1477, by Bio-Rad (1 mentions) Ab15277, by Abcam (1 mentions) Ab9610, by R&D Systems (1 mentions) Mab2018, by R&D Systems (1 mentions) Af1062, by R&D Systems (1 mentions) Rabbit anti histone h3, by Santa Cruz Biotechnology (1 mentions) Rabbit anti gfap, by Agilent Technologies (1 mentions) Eclipse ci microscope, by Nikon (1 mentions) Ab75769, by Abcam (1 mentions)

15 protocols using a21240

Immunofluorescent Staining of Tissue Sections

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Slides were deparaffinized through a graded alcohol series, and antigen retrieval was performed using a decloaking chamber (Biocare medical, #DC2002) at 125°C for 3 minutes using Diva buffer (Biocare medical, #DV2004G1). Slides were blocked for 1 hour using 0.2% bovine serum albumin + 2% normal goat serum, incubated with primary antibodies (CD3 clone CD3-12, bio-rad #MCA1477, Granzyme B, rabbit polyclonal, LSBio # 34084), diluted 1:500 each for 1 hour in blocking buffer for 1 hour, RT. Slides were washed in PBS, and incubated in secondary antibody (Fisher #A-21240) diluted 1:500 in blocking buffer for 1 hour, RT. Slides were washed in ddH2O, and incubated with Hoechst dye (Fisher, #H3569) diluted to 2 μg/ml in ddH2O for 10 minutes. Slides were imaged on a Nuance Multispectral Imaging system on a Nikon Eclipse Ci microscope.

Ravanpay A.C., Gust J., Johnson A.J., Rolczynski L.S., Cecchini M., Chang C.A., Hoglund V.J., Mukherjee R., Vitanza N.A., Orentas R.J, & Jensen M.C. (2019). EGFR806-CAR T cells selectively target a tumor-restricted EGFR epitope in glioblastoma. Oncotarget, 10(66), 7080-7095.

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Antibody Characterization for Cellular Analysis

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The following primary antibodies were used at indicated dilutions: mouse anti-APC/CC1 [immunohistochemistry (IHC) 1:20; Millipore OP80], mouse anti-CNP [western blotting (WB) 1:1000; Millipore C5922], rabbit anti-Dystrophin (WB 1 µg/ml, Abcam AB15277), mouse anti-beta-dystroglycan (WB 1:250; Developmental Studies Hybridoma Bank), rat anti-Ki67 (IHC: 1:100, Invitrogen 4-5698-82), rat anti-MBP (IHC 1:200; WB 1:1000; BioRad AA82-87), rabbit anti-Histone H3 (1:750; Santa Cruz), rabbit anti-MOG (WB: 1:1000; Abcam ab108505), rabbit anti-GFAP (1:1000; DAKO), rabbit anti-Olig2 (IHC 1:100; Millipore AB9610), goat anti-PDGFRα , (IHC 1:100; R&D Systems AF1062), mouse anti-SOX2 (IHC 1:100; R&D Systems MAB2018), mouse (IgG1) anti-β-Catenin [immunofluorescence (IF) Wholemount 1:500; BD Transduction Labs 610153], rabbit anti-γ-Tubulin (IF Wholemount 1:500; Sigma T5192). Olig2 was visualized using anti-rabbit CY5, Ki67 was visualized using anti-rat Alexa-Fluor 488, Sox2 was visualized using anti-mouse CY3, and PDGFRα was visualized using antigoat Alexa-Fluor 680. The following secondary antibodies were used at indicated dilutions: anti-mouse IgG1 (IF Wholemount 1:500; Fisher A-21240), anti-rabbit IRDye 800 Donkey (1:2500; Licor AB_2715510), anti-mouse IRDye 800 Donkey (1:2500; Licor AB_621847), anti-rat (IRDye 800 Goat) (1:4000; Licor AB_2721932).

Arreguin A.J., Shao Z, & Colognato H. (2024). Dmdmdx mice have defective oligodendrogenesis, delayed myelin compaction and persistent hypomyelination. Disease Models & Mechanisms, 17(4), dmm050115.

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Immunohistochemical Staining of Tissue Samples

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Slides were deparaffinized through a graded alcohol series, and antigen retrieval was performed using Digest-all 3 enzyme cocktail (Fisher, #3009) for 10 minutes at 37°C. Slides were blocked for 1 hour using 0.2% bovine serum albumin + 2% normal goat serum, incubated with primary antibody (clone 31G7, Fisher, #280005), diluted 1:100 for 1 hour in blocking buffer for 1 hour, RT. Slides were washed in PBS, and incubated in secondary antibody (Fisher #A-21240) diluted 1:500 in blocking buffer for 1 hour, RT. Slides were washed in ddH2O, and incubated with Hoechst dye (Fisher, #H3569) diluted to 2 μg/ml in ddH2O for 10 minutes. Slides were imaged on a Nuance Multispectral Imaging system on a Nikon Eclipse Ci microscope.

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Immunostaining of Malaria Gametocytes and Bone Cells

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Ten-μm-thick sections were air-dried, hydrated with PBS and exposed to 5% goat serum in PBS for 30 minutes at RT. For immunostaining of gametocytes, the sections were incubated with a mouse monoclonal anti-Pfs16 antibody (Bruce et al., 1994 (link)) (32F717-SF, kindly provided by Dr. R. Sauerwein, Radboud University Medical Centre, Nijmegen, The Netherlands), at a dilution of 1:500 in PBS, O/N at 4°C. For immunostaining of cells within the ossicles the following antisera were used: rabbit polyclonal anti-mouse and human Fatty Acid Binding Protein (FABP4, ab13979, Abcam Cambridge, UK), anti-human CD146 (ab75769, Abcam), anti-human Alkaline Phosphatase (ALP, 11187-1-AP, ProteinTech Manchester, UK) all applied at 1:100 in PBS, O/N at +4°C. Following washing with PBS, samples were incubated with Alexa Fluor 647-conjugated goat anti-mouse (A-21240, Thermo Fisher Scientific) or Alexa Fluor 488-conjugated goat anti-rabbit (A-11008, Thermo Fisher Scientific) at dilution of 1:200 in PBS for 1 hr at RT. Nuclei were counterstained with TO-PRO-3.

Donsante S., Siciliano G., Ciardo M., Palmisano B., Messina V., de Turris V., Farinacci G., Serafini M., Silvestrini F., Corsi A., Riminucci M, & Alano P. (2023). An in vivo humanized model to study homing and sequestration of Plasmodium falciparum transmission stages in the bone marrow. Frontiers in Cellular and Infection Microbiology, 13, 1161669.

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Immunofluorescence Analysis of Mitochondria

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U937 cells were treated with 50nM PMA as described above. On the third day after trypsinization to remove the PMA, 70-100,000 cells were plated onto glass coverslips in 24-well plates with 1ml fresh media. Two days later, cells were washed with 1x PBS and fixed using 4% formaldehyde for 10 minutes. After fixation, cells were washed with 1x PBS, permeabilized with 0.1% Triton X-100, blocked with 5% normal goat serum, and stained with the following antibodies: mouse monoclonal anti-CoxIV (1:1000, Abcam), rabbit polyclonal anti-FIS1 (1:200, Proteintech 10956), and rabbit polyclonal anti-TBC1D15 (1:100, Sigma-Aldrich HPA013388). Goat anti-rabbit Alexa Fluor 488 (1:1000, Thermo Fisher Scientific A-11034) and goat anti-mouse Alexa Fluor 647 (1:1000, Thermo Fisher Scientific A-21240) secondary antibodies were used for visualization. Coverslips were mounted using Prolong Diamond Antifade Mountant with DAPI (Thermo Fisher Scientific). Images were acquired using a Leica DMI6000B inverted fluorescence microscope with a 100X oil immersion objective.

Chai N., Haney M.S., Couthouis J., Morgens D.W., Benjamin A., Wu K., Ousey J., Fang S., Finer S., Bassik M.C, & Gitler A.D. (2019). Genome-wide synthetic lethal CRISPR screen identifies FIS1 as a genetic interactor of ALS-linked C9ORF72. Brain research, 1728, 146601.

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Immunofluorescence Staining of Cellular Proteins

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Cells plated on glass coverslips were fixed with 4% paraformaldehyde and blocked with 3% BSA in phosphate-buffered saline (PBS) for 1 hour at room temperature. Primary antibodies were incubated overnight at 4°C with dilutions as follows: mouse anti-LAMP2 (1:500; H4B4, Santa Cruz Biotechnology, RRID: AB_626858) and rabbit anti–hemagglutinin (HA) (1:500; C29F4, Cell Signaling Technology, RRID: AB_1549589). Secondary antibodies (Alexa Fluor) were then applied in 1% BSA and 0.3% Triton X-100 for 3 hours. The following secondary antibodies were used (1:1000): goat anti-mouse Alexa Fluor 647 (A-21240, Thermo Fisher Scientific, RRID: AB_2535809) and goat anti-rabbit Alexa Fluor 594 (A32740, Thermo Fisher Scientific, RRID: AB_2762824). Confocal images were acquired on the Zeiss LSM980 with Airyscan 2.

Scharenberg S.G., Dong W., Ghoochani A., Nyame K., Levin-Konigsberg R., Krishnan A.R., Rawat E.S., Spees K., Bassik M.C, & Abu-Remaileh M. (2023). An SPNS1-dependent lysosomal lipid transport pathway that enables cell survival under choline limitation. Science Advances, 9(16), eadf8966.

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Immunohistochemical Analysis of Lung Tissues

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Isolated lungs were fixed in 4% paraformaldehyde in PBS for 30 minutes at 4°C. Lungs were washed four times for 15 minutes with PBST (0.1% Triton X-100 in PBS) and then blocked with 5% goat serum and 0.1% BSA for one hour. Samples were then incubated with primary antibodies against αSMA (Sigma a5228, 1:400 or Abcam ab5694, 1:200), E-cadherin (Cell Signaling 3195, 1:200 or Invitrogen 13–1900, 1:200), fibronectin (Invitrogen PA5-29578, 1:200), Lef1 (Cell Signaling 2230, 1:200), Ki67 (Abcam a15580, 1:200), pMLC2 (Cell Signaling 3671, 1:200), RFP (Abcam ab62341, 1:400), Sox9 (Sigma AB5535, 1:1000), or Tbx3 (Invitrogen 42–4800, 1:200), followed by incubation with Alexa Fluor-conjugated secondary antibodies (1:400; Thermo Fisher Scientific A11007, A21244, A11012, A21240 and A11006) or Alexa Fluor-conjugated phalloidin (1:400, Thermo Fisher Scientific A12380 or A12379) and then counterstained with Hoechst (1:1000). Lungs were dehydrated in a methanol or isopropanol (for phalloidin-stained samples) series and cleared with Murray’s clear (1:2 ratio of benzyl alcohol to benzyl benzoate). Samples were imaged using a spinning disk confocal (BioVision X-Light V2) fitted to an inverted microscope.

Schönbrunner E.R, & Schmid F.X. (1992). Peptidyl-prolyl cis-trans isomerase improves the efficiency of protein disulfide isomerase as a catalyst of protein folding.. Proceedings of the National Academy of Sciences of the United States of America, 89(10), 4510-4513.

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Immunostaining of Differentiated Nasal and Bronchial ALI Cultures

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Differentiated nasal and bronchial ALI cultures (n = 3) were fixed and stained as previously described [25 (link)]. Primary antibodies (β-tubulin IV (MU178-UC, Emergo Biogenex, Fremont, CA, USA), MUC5AC (ab198294, Abcam, Cambridge, UK), were incubated for 1 h at RT. Afterward, secondary antibodies (A-21240 and A11034, Invitrogen, Waltham, MA, USA), together with phalloidin (A34055, Mol. Probes, Eugene, OR, USA) and DAPI (D9542, Sigma, St. Louis, MO, USA), were added for 30 min. Both primary and secondary antibodies were diluted 1:500 in blocking buffer (1% BSA + 0.3 Triton X-100 (T8787, Sigma-Aldrich, St. Louis, MO, USA) in PBS). Images were acquired with a Zeiss LSM800 confocal microscope (40× objective). Image quantification was performed using Fiji (Max Planck Institute, Dresden, Germany) and CellProfiler (Broad Institute, Cambridge, MA, USA). Cultures did not show contamination with other cell types such as fibroblasts.

Dreyer H.H., van Tuyll van Serooskerken E.S., Rodenburg L.W., Bittermann A.J., Arets H.G., Reuling E.M., Verweij J.W., Haarman E.G., van der Zee D.C., Tytgat S.H., van der Ent C.K., Beekman J.M., Amatngalim G.D, & Lindeboom M.Y. (2023). Airway Epithelial Cultures of Children with Esophageal Atresia as a Model to Study Respiratory Tract Disorders. Children, 10(6), 1020.

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Immunostaining of Phosphorylated EGFR

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EGFR antibody (clone H11, DAKO, Amstelveen, the Netherlands) and a phospho-specific EGFR antibody (AB32430, anti phospho Y1068, Abcam, Cambridge, UK) were used at 1:500 dilution for both western blot and immunohistochemistry. Secondary antibodies used were Alexafluor 647 goat anti-mouse (A21240, Invitrogen, Bleiswijk, the Netherlands) and Alexafluor 488 goat anti-rabbit (A11008, Invitrogen, Bleiswijk, the Netherlands). Hoechst and WGA were used as counterstain to visualize nucleus and membranes respectively.

de Wit M., Gao Y., Mercieca D., de Heer I., Valkenburg B., van Royen M.E., Aerts J., Sillevis Smitt P, & French P. (2020). Mutation and drug-specific intracellular accumulation of EGFR predict clinical responses to tyrosine kinase inhibitors. EBioMedicine, 56, 102796.

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Immunohistofluorescence of Liver Fibrosis

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Formalin fixed and paraffin embedded liver fibrosis tissue arrays were purchased from US Biomax (LV805a) (Rockville, MD). Tissue array patient information is shown in Supplementary Table 2. IHF was performed according to a previous report34 (link). Thin cryosections (4 μm) of liver tissue were fixed in acetone for immunohistofluorescence, stained with the indicated antibodies. Antibodies used in IHF were: anti-CUGBP1 (Santa Cruz Biotechnology, SC-20003, 1: 50), α-SMA (Santa Cruz Biotechnology, SC-32251, 1: 50), Reelin (Abcam, ab78540, 1: 50), IFN-γ (Abcam, ab133566, 1: 50), Cytoglobin (Abcam, ab57713, 1: 50), CUGBP1 conjugated to Alexa Fluor 488 (Abcam, ab129115, 1: 100), IFN-γ conjugated to PE-CF594 (BD Biosciences, 562303, 1: 50), goat anti-mouse IgG2a conjugated to Alexa Fluor 594 (Invitrogen, A-21135, 1: 500), goat anti-mouse IgG1 conjugated to Alexa Fluor 647 (Invitrogen, A-21240, 1: 500), goat anti-rabbit IgG conjugated to Alexa Fluor 594 (Invitrogen, R37117, 1: 500), goat anti-mouse IgG2b conjugated to Alexa Fluor 488 (Invitrogen, A-21141, 1: 500). The sections were then stained with DAPI and examined with a confocal laser scanning microscope (Leica, Wetzlar, Germany). Sirius Red staining was performed by Servicebio (Wuhan, China). The liver fibrosis stage was assessed by Ishak scale35 (link).

Wu X., Wu X., Ma Y., Shao F., Tan Y., Tan T., Gu L., Zhou Y., Sun B., Sun Y., Wu X, & Xu Q. (2016). CUG-binding protein 1 regulates HSC activation and liver fibrogenesis. Nature Communications, 7, 13498.

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