Fluorchem q

Manufactured by Bio-Techne

Sourced in United States, United Kingdom

The FluorChem Q is a compact and versatile fluorescence imaging system designed for a wide range of applications in life science research. It features a high-resolution CCD camera, multiple fluorescence and visible light excitation sources, and advanced image acquisition and analysis software. The FluorChem Q enables sensitive and quantitative imaging of fluorescent samples, including protein gels, cell-based assays, and DNA/RNA analysis.

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Lab products found in correlation

Pierce bca protein assay kit, by Thermo Fisher Scientific (8 mentions) Protran ba83, by Cytiva (3 mentions) Protease inhibitor cocktail, by Roche (3 mentions) Pvdf membrane, by Bio-Rad (3 mentions) Exoab cd63a 1, by System Biosciences (3 mentions) Hsc70, by Santa Cruz Biotechnology (2 mentions) Total acc, by Cell Signaling Technology (2 mentions) Ripa lysis buffer, by Merck Group (2 mentions) Protran ba85, by GE Healthcare (2 mentions) Alphaview software, by Bio-Techne (2 mentions) Supersignal west pico kit, by Thermo Fisher Scientific (2 mentions) Pierce micro bca protein assay kit, by Thermo Fisher Scientific (2 mentions) Sc 543x, by Santa Cruz Biotechnology (2 mentions) Tgx protein gels, by Bio-Rad (2 mentions) Rabbit anti β actin, by Cell Signaling Technology (2 mentions) Anti rabbit igg hrp linked antibody, by Cell Signaling Technology (2 mentions) Ecl western blotting detection reagent, by GE Healthcare (2 mentions) Tween 20, by Thermo Fisher Scientific (2 mentions) Bca protein assay, by Thermo Fisher Scientific (2 mentions) Acsl1, by Cell Signaling Technology (1 mentions)

Spelling variants of this product

FluorChem Q imager (10 citations) FluorChem Q System (10 citations) Proteinsimple+Fluorchem Q system (5 citations) MultiImage III FluorChem Q (5 citations) FluorChem Q Imaging software (4 citations) FluorChem Q imaging analysis system (3 citations) Proteinsimple + Fluorchem Q (2 citations)

21 protocols using fluorchem q

Liver and Kidney Protein Expression

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Liver and kidney homogenates were prepared using 1× RIPA buffer with protease inhibitors. The protein concentration was measured using the Pierce BCA Protein Assay kit (Thermo Scientific). We used 30 ug of protein on an SDS-PAGE and then transferred it to either a nitrocellulose (Protran BA 83, Whatman) or a polyvinylidene difluoride (PVDF) membrane. We then blocked with 3% BSA-TBST (tris buffer saline with tween 20). The membranes were then probed with antibodies Cpt2 (Pierce), Pgc1α (Abcam), Acsl1 (Cell Signaling), Acot1 (Cell Signaling), Acot2 (Cell Signaling), Cpt1α (Abcam), Acadm (GeneTex), Acsf3 (Pierce), Total Acc (Cell Signaling), Acly (Cell Signaling), Hadha (Genetex), Aco2 (Cell Signaling), Fasn (BD Biosciences), and Hsc70 (Santa Cruz Biotechnology). Hsc70 used the appropriate Cy3 fluorescent secondary antibodies, and the other primary antibodies used the corresponding secondary antibodies conjugated to horseradish peroxidase. Images were collected using an Alpha Innotech FluorChemQ and presented with minimal image processing.

Lee J., Choi J., Scafidi S, & Wolfgang M.J. (2016). Hepatic fatty acid oxidation restrains systemic catabolism during starvation. Cell reports, 16(1), 201-212.

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Western Blot Analysis of Adipose Tissue

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BAT, iWAT, and gWAT depots were homogenized with 300–500 ul of RIPA buffer (50 mM Tris–HCl, pH 7.4, 150 mM NaCl, 1 mM EDTA, 1% Triton X-100, 0.25% deoxycholate) containing protease inhibitor cocktail (Roche) and phospho-STOP cocktail (Roche), followed by pelleting of the insoluble debris at 13,000×g for 15 min at 4 °C. The protein concentrations of lysates were determined by BCA assay (Thermo Scientific), and 30 μg of lysate was separated by Tris-glycine SDS-PAGE (10% polyacrylamide). Proteins were transferred to nitrocellulose membranes (Protran BA 83, Whatman), blocked in 3% BSA in 1X TBST (Tris-buffered saline with Tween 20), and incubated with primary antibodies overnight. The blots were probed with the following antibodies: Ucp1 (Sigma; U6382), Ndufb8, Sdhb, Uqcrc2, Atp5a (MitoProfile total OXPHOS, Abcam; ab110413), Aco2 (Cell Signaling; 6922), Mcad (GeneTex; GTX32421), Pcx (Abcam; ab128952), Vdac (Calbiochem; PC548), Pdh E2/E3bp (Abcam; ab110333), Acot2 (Sigma; SAB2100030), beta-Actin (Sigma; A2228), and Hsc-70 (Santa Cruz; sc-7298). Cy3-conjugated anti-mouse (Invitrogen), or HRP-conjugated anti-mouse (GE Healthcare) or anti-rabbit (GE Healthcare) secondary antibodies were used appropriately. Images were collected and analyzed using an Alpha Innotech FluorChemQ.

Gonzalez-Hurtado E., Lee J., Choi J, & Wolfgang M.J. (2017). Fatty acid oxidation is required for active and quiescent brown adipose tissue maintenance and thermogenic programing. Molecular Metabolism, 7, 45-56.

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Detecting EV Proteins by Western Blot

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Protein from neurospheres and isolated EVs was extracted using 1xRIPA lysis buffer (EMD Millipore; Reference # 20–188), with addition of Halt protease & phosphatase inhibitor cocktail (Thermo Fisher Scientific; Product # 78442). Extracted protein concentration was determined using Pierce BCA protein assay kit (Thermo Fisher Scientific; Catalog # 23225). 25ug of protein was size-fractionated on a 4–12% Bis-Tris Gel, ran at 150V for 60 minutes, and blotted to a PVDF membrane using iBlot transfer system (Thermo Fisher Scientific; Catalog # IB301001). Subsequently, the membrane was blocked for one hour with 2.5% goat serum and 5% nonfat dry milk in Tris-buffered saline that contains 0.1% Tween-20 (TTBS), then incubated overnight with polyclonal rabbit anti-CD63 antibodies diluted 1:1000 (System Biosciences; Catalog # EXOAB-CD63A-1). CD63 is an established surface marker for EVs. The blot was washed and incubated with an HRP-conjugated goat anti-rabbit IgG (Invitrogen) at dilution 1:1000 for one hour, then developed using PerkinElmer Western Lightning Plus-ECL (PerkinElmer; Catalog # NEL103001EA) and visualized using a CCD camera (Fluorchem Q, Alpha Innotech).

Tseng A.M., Chung D.D., Pinson M.R., Salem N.A., Eaves S.E, & Miranda R.C. (2019). Ethanol Exposure Increases miR-140 in Extracellular Vesicles: Implications for Fetal Neural Stem Cell Proliferation and Maturation. Alcoholism, clinical and experimental research, 43(7), 1414-1426.

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Anchorage-Independent Cell Growth Assay

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Cells were plated on a solidified bed of 0.5% agarose in growth medium in 3.5-cm plates. For plating, 1 × 10⁴ 3T3–EGFR-WT, 3T3–EGFR-R776G, 3T3–EGFR-R776H, or 3T3–BRAF-V600E stable cell lines were resuspended in 0.35% agarose in growth medium. Agar was topped with 2 ml of growth medium and incubated overnight. Twenty-four hours after plating, top medium was removed and replaced with pHi control medium (“Control,” DMEM + 10% FBS; “NH₄Cl, ” DMEM + 10% FBS + 5 mM NH₄Cl). For BRAF plates, BRAF-V600E expression was induced with 1 µM tamoxifen. Medium was replaced every 3 days until day 15, when cells were washed twice in PBS, fixed with 4% paraformaldehyde in PBS for 1 hour, washed twice in PBS, and stained with 0.0005% crystal violet for 2 hours. After PBS washes to remove excess crystal violet, images were acquired (Alpha Innotech FluorChem Q). Colonies were counted using Analyze Particles function on ImageJ software. Particles larger than 8 pixel² were quantified and averaged from two technical replicates per condition.

White K.A., Ruiz D.G., Szpiech Z.A., Strauli N.B., Hernandez R.D., Jacobson M.P, & Barber D.L. (2017). Cancer-associated arginine-to-histidine mutations confer a gain in pH sensing to mutant proteins. Science signaling, 10(495), eaam9931.

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Analyzing Hepatic Lipid Metabolism Proteins

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Liver homogenates from 16-week HFD or acute insulin injected Cpt2 ^lox/lox and Cpt2^L−/− mice were prepared using 1x RIPA buffer with protease and phosphatase inhibitors. The protein concentration was measured using the Pierce BCA Protein Assay kit (Thermo Scientific). We used 30μg of protein on an SDS-PAGE and then transferred it to a nitrocellulose (Protran BA 83, Whatman) membrane. We then blocked with 3% BSA-TBST (Tris-buffered saline with Tween20). The membranes were probed with antibodies Pcx (Cell Signaling), Acly (Cell Signaling), Total Acc (Cell Signaling), Fasn (BD Biosciences), Scd1 (Cell Signaling), Hsp27 (Cell Signaling), Mitoprofile (Abcam) and Hsc70 (Santa Cruz Biotechnology) diluted 1:1000 with 3% BSA in TBST. Hsc70 used the appropriate Cy3 fluorescent secondary antibody, and the other primary antibodies used corresponding secondary antibodies conjugated to horseradish peroxidase. Images were collected using an Alpha Innotech FluorChemQ and presented with minimal image processing.

Lee J., Choi J., Selen Alpergin E.S., Zhao L., Hartung T., Scafidi S., Riddle R.C, & Wolfgang M.J. (2017). Loss of hepatic mitochondrial long chain fatty acid oxidation confers resistance to diet-induced obesity and glucose intolerance. Cell reports, 20(3), 655-667.

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Western Blot Analysis of Optic Nerve Proteins

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Aliquots of optic nerve lysate and immunoprecipitated proteins were loaded into 4–12% gradient gel lanes (Bis-Tris, Invitrogen), electrophoretically separated in MOPS running buffer at 200 V for 1 h, and transferred to polyvinyl difluoride (PVDF) membranes (Bio-Rad) at 30 V for 1 h. Protein standards (SeeBlue Plus2) were run in lanes adjacent to the samples. Each PVDF membrane was immersed for 1 h in a protein-supplemented buffer solution (Superblock), and then incubated with primary antibodies on a rocker overnight at 4°C. As a loading control, lanes were also immunostained for either β-actin or myelin basic protein. The membranes were then rinsed with Tris-buffered saline (supplemented with 0.5% Tween 20) and incubated in species-specific, fluorophore-conjugated secondary antibodies for 1 h at room temperature. The fluorescence of these fluorophores was visualized and imaged in a digital imager (FluorChem Q, Alpha Innotech; RRID:SCR_014549). The molecular weights of the immunostained protein bands were analyzed by use of the imager software (AlphaView Q, ProteinSimple). Unless stated otherwise, measurements are presented as the mean ± S.E.M., plus sample size.

Ogata G., Partida G.J., Fasoli A, & Ishida A.T. (2022). Calcium/calmodulin-dependent protein kinase II associates with the K+ channel isoform Kv4.3 in adult rat optic nerve. Frontiers in Neuroanatomy, 16, 958986.

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Protein Expression Analysis in Cancer Cells

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After 36 h transfection, 769-P and OS-RC-2 cells were treated with RIPA lysis buffer for 40 min on ice. The protein concentration was determined using a BCA protein assay kit (Bio-Rad Laboratories, Hercules, CA, USA), as described in the manufacturer's manual. Lysate proteins (40 µg) were subjected to 10% SDS-PAGE and electrophoretically transferred to polyvinylidene difluoride (PVDF) membranes. After blotting, the membranes were blocked with 5% fat-free dry milk for 1 h and then incubated with the specific primary antibody against eIF4E, cyclin D1, c-Myc, MMP3, Bcl-2, P13K, p-AKT, mTOR or β-actin (Santa Cruz Biotechnology, Santa Cruz, CA, USA) for 2 h at 37°C. After rinsing, membranes were hybridized with HRP-conjugated secondary antibodies for 1 h at 37°C. The protein bands were detected using a quantitative gel and a Western Blot Imaging System (FluorChem Q; Alpha Innotech, San Leandro, CA, USA).

Li G., Chong T., Xiang X., Yang J, & Li H. (2017). Downregulation of microRNA-15a suppresses the proliferation and invasion of renal cell carcinoma via direct targeting of eIF4E. Oncology Reports, 38(4), 1995-2002.

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Neurosphere Protein Extraction and Western Blot

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Cultured neurospheres were collected by centrifugation and lysed in 1X RIPA lysis buffer (EMD Millipore, Burlington, MA) supplemented with Halt protease inhibitor cocktail (Thermo Fisher Scientific). Protein concentration was determined using Pierce BCA protein assay kit (Thermo Fisher Scientific). Cleared cell lysates were size fractionated on a 4-12% Bis-Tris gradient acrylamide gel (Thermo Fisher Scientific, Rockford, IL), at 150 volts for 60 min, and transferred to a PVDF membrane using the iBlot dry transfer system and manufacturer protocols (Thermo Fisher Scientific). The membrane was then blocked with either 5% nonfat dry milk and 2.5% goat serum in Tris-buffered saline containing Tween 20 (TTBS) or Odyssey® Blocking Buffer (LI-COR Biotechnology. Lincoln, Nebraska) in TBS for 1 hour, incubated overnight with primary antibody, washed and incubated with secondary antibody for 1 hour at room temperature (list of antibodies and dilutions are presented in Supplementary File 1g), and subsequently developed either using PerkinElmer Western Lightning Plus Chemi-ECL (PerkinElmer, Waltham, MA) and visualized using a CCD camera (Fluorchem Q, Alpha Innotech, San Leandro, CA) or direct visualization of near infrared-conjugated secondary antibodies using Odyssey® CLx Imaging System (LI-COR Biotechnology. Lincoln, Nebraska).

Salem N.A., Mahnke A.H., Tseng A.M., Garcia C.R., Jahromi H.K., Geoffroy C.G, & Miranda R.C. (2020). A novel Oct4/Pou5f1-like non-coding RNA controls neural maturation and mediates developmental effects of ethanol.. Neurotoxicology and teratology, 83, 106943.

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Chemiluminescent Visualization of PelC and Psl

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Chemiluminescent Western and dot blots for PelC and Psl, respectively, were carried out by established methods [80 (link)] with slight modification (see S1 Text). Horse-radish peroxidase linked secondary antibodies were used in conjunction with the Pierce SuperSignal West Pico ECL reagent (Thermo Scientific) to visualize proteins bound by primary antibodies and Chemiluminescence was captured and quantified using a FluorChem Q (Alpha Innotech).

Harrison J.J., Almblad H., Irie Y., Wolter D.J., Eggleston H.C., Randall T.E., Kitzman J.O., Stackhouse B., Emerson J.C., Mcnamara S., Larsen T.J., Shendure J., Hoffman L.R., Wozniak D.J, & Parsek M.R. (2020). Elevated exopolysaccharide levels in Pseudomonas aeruginosa flagellar mutants have implications for biofilm growth and chronic infections. PLoS Genetics, 16(6), e1008848.

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Western Blot Analysis of Ubiquitinated β-Catenin

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Proteins were transferred to 0.45-µm PVDF membranes (100 V for 1 h on ice; Immobilon) in modified transfer buffer (Tris-Gly, 20% MeOH, and 0.01% SDS) to maximize transfer of ubiquitinated β-catenin. Membranes were blocked in 5% BSA in TBS-T (0.1% Tween-20 in TBS) for 1 h at RT and then divided for blotting. Primary antibodies: total β-catenin (1:1,000; 610154; BD), pSer (33/37) pThr41 β-catenin (1:1,000; 9561; Cell Signaling Technology), pSer45 β-catenin (1:1,000; 9564; Cell Signaling Technology), and β-TrCP (1:1,000; D13F10; 4349; Cell Signaling Technology), ubiquitin (1:2,000; 3933; Cell Signaling Technology), and E-cadherin (1:2,000; 610181; BD). Primary antibodies were used at indicated dilutions in 5% BSA/TBS-T and incubated overnight with shaking at 4°C. Membranes were washed with shaking (3 × 5 min; TBS-T). Secondary HRP-conjugated antibodies against mouse or rabbit (1706515 and 1721019; Bio-Rad; 1:10,000 in 5% BSA/TBS-T) were incubated for 1 h (RT with shaking). Membranes were washed with shaking (3 × 5 min; TBS-T), developed (SuperSignal West Femto; Pierce), and visualized (Alpha Innotech FluorChem Q).

White K.A., Grillo-Hill B.K., Esquivel M., Peralta J., Bui V.N., Chire I, & Barber D.L. (2018). β-Catenin is a pH sensor with decreased stability at higher intracellular pH. The Journal of Cell Biology, 217(11), 3965-3976.

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