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Fluorchem 8800

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The FluorChem 8800 is a digital imaging system designed for analyzing and quantifying fluorescent and chemiluminescent samples. It features a high-resolution CCD camera, multiple excitation and emission filters, and advanced imaging software for capturing and analyzing images.

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Lab products found in correlation

Dc protein assay kit, by Bio-Rad (3 mentions) Protease and phosphatase inhibitor cocktail, by Merck Group (2 mentions) Horseradish peroxidase conjugated secondary antibody, by Cell Signaling Technology (1 mentions) Trans blot turbo, by Bio-Rad (1 mentions) β actin, by Merck Group (1 mentions) T per solution, by Thermo Fisher Scientific (1 mentions) Laemmli sample buffer, by Bio-Rad (1 mentions) Enhanced chemiluminescence detection system, by GE Healthcare (1 mentions) Hrp conjugated goat anti rabbit igg, by Merck Group (1 mentions) Rabbit anti dnp antibody, by Merck Group (1 mentions) Blocking agent, by GE Healthcare (1 mentions) Chemiluminescent substrate kit, by GE Healthcare (1 mentions) Hybond p, by GE Healthcare (1 mentions) Rabbit anti gapdh antibody, by Merck Group (1 mentions) Rabbit anti bdnf, by Abcam (1 mentions) Rnalater, by Thermo Fisher Scientific (1 mentions) Odyssey infrared imaging system, by LI COR (1 mentions) Anti β actin, by Merck Group (1 mentions) T per buffer, by Thermo Fisher Scientific (1 mentions) Methocult h4230, by STEMCELL (1 mentions)

Close spelling variants of this product

FluorChem 8800 Image Detection System FluorChem IS-8800 Imager

8 protocols using fluorchem 8800

1

Phospho-CREB Quantification in Ganglia

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Ganglia were homogenized in T-Per solution (Pierce Biotechnology, Rockford, IL) with addition of protease and phosphatase inhibitor cocktails (Sigma). Proteins were denatured in an equivalent amount of Laemmli Sample Buffer (Bio-Rad) and were separated on a 10% SDS-PAGE gel and transferred to a nitrocellulose membrane using Trans-Blot Turbo (Bio-Rad). The membrane was blocked using 5% milk in Tris-buffered saline, and incubated at 4℃ with primary antibody against phospho-CREB (1:1000, Cell Signaling). A horseradish peroxidase-conjugated secondary antibody (1:2000, Cell Signaling) and enhanced chemiluminescence system was used to visualize immunoreactive bands. For endogenous control, the same membrane was stripped and re-incubated with antibody against total CREB (1:1000, Cell Signaling), or β-actin (1:5000, Sigma). The densitometric quantification of immunoreactive bands was performed using the software FluorChem 8800 (Alpha Innotech, San Leabdro, CA).
Hashmi F., Liu M., Shen S, & Qiao L.Y. (2016). Phospholipase C gamma mediates endogenous brain-derived neurotrophic factor-regulated calcitonin gene-related peptide expression in colitis-induced visceral pain. Molecular Pain, 12, 1744806916657088.
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2

Quantification of Protein Carbonylation

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Protein carbonyls were measured as described (Levine et al. 2000 (link)) and modified by Wen et al. (2006a (link), 2008 (link)). Briefly, 20 μg proteins was denatured and derivatized in 3 % sodium dodecyl sulfate (SDS), 10 mM 2,4-dinitrophenylhydrazine (DNPH) dissolved in 10 % trifluoroacetic acid (20-μl volume). After neutralization with 7.5 μl of 2 M Tris, 30 % glycerol, DNP-derivatized protein samples were resolved by 10 % of SDS–PAGE. Gels were transferred to PVDF membranes. Carbonylized proteins were probed with rabbit anti-DNP antibody (1:2,000, Sigma-Aldrich, St Louis, MO), followed by HRP-conjugated goat anti-rabbit IgG (1:5,000, Sigma-Aldrich), and signal was developed by using an enhanced chemiluminescence detection system (GE Healthcare, Pittsburgh, PA). Images were visualized, digitized, and quantified by densitometry using a FluorChem 8800 (Alpha Innotech, San Jose, CA) image analyzing system.
Wen J.J., Nagajyothi F., Machado F.S., Weiss L.M., Scherer P.E., Tanowitz H.B, & Garg N.J. (2014). Markers of oxidative stress in adipose tissue during Trypanosoma cruzi infection. Parasitology research, 113(9), 3159-3165.
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3

Western Blot Analysis of Signaling Proteins

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The protein extracts from pooled L1–L2 and L6–S1 DRGs were subject to centrifugation at 20,200 g for 10 min at 4 °C, and the supernatant was removed to a fresh tube. The protein concentration was determined using Bio-Rad DC protein assay kit. Proteins were then separated on a 10 % SDS-PAGE gel and transferred to a nitrocellulose membrane. The membrane was blocked with 5 % milk in Tris-buffered saline for 1 h and then incubated with a specific primary antibody followed by HRP-conjugated secondary antibody. For internal loading control, the same membrane was stripped and re-probed with anti-β-actin antiserum or antibody to a non-phosphorylated form of the protein examined. The concentrations of antibodies used for p-CREB, CREB, p-Akt, Akt, PLCγ and p-CaMKII were 1:1000, and for β-actin was 1:3000 (Sigma-Aldrich, St. Louis, MO). The bands were visualized by enhanced chemiluminescence (ECL). Densitometric quantification of the immunoreactive bands was performed using the software FluorChem 8800 (Alpha Innotech, San Leandro, CA).
Xia C., Shen S., Hashmi F, & Qiao L.Y. (2015). Colitis-induced bladder afferent neuronal activation is regulated by BDNF through PLCγ pathway. Experimental neurology, 285(Pt B), 126-135.
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4

Quantifying Hippocampal Stress Proteins

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For Western blot analysis, collected hippocampi were placed in RNAlater (Life Technologies) and homogenized in an SDS sample buffer. Protein extracts were separated by SDS–polyacrylamide gel electrophoresis and then transferred onto a polyvinylidene difluoride membrane (Hybond P; GE Healthcare). The membrane was blocked with a blocking agent (GE Healthcare) and then incubated at 4°C overnight with a primary antibody for rabbit anti-HSP110 (1:5000, StressMarq Biosciences Inc.), rabbit anti-HSP25 (1:5000, Cosmo Bio Co. Ltd.), mouse anti-HSP60 (1:1000, Cosmo), mouse anti-HSP72 (1:1000, Cosmo), mouse anti–phospho-CREB (Ser133) (1:5000, Cell Signaling Technology), rabbit anti-CREB (Ser133) (1:5000, Cell Signaling), and rabbit anti-BDNF (1:5000, Abcam). After washing with tris-buffered saline containing 0.1% (v/v) Tween 20, the membranes were incubated with horseradish peroxidase–conjugated secondary antibody (1:20,000) for 1 hour at room temperature. The antibody-reactive bands were visualized using the chemiluminescent substrate kit (GE Healthcare). Bands were analyzed by densitometry using FluorChem 8800 (Alpha Innotech), and the content of GAPDH, which was detected using a rabbit anti-GAPDH antibody (1:20,000; Sigma), was used as a control to ensure that the same amount of protein was loaded in each lane.
Hashikawa N., Utaka Y., Ogawa T., Tanoue R., Morita Y., Yamamoto S., Yamaguchi S., Kayano M., Zamami Y, & Hashikawa-Hobara N. (2017). HSP105 prevents depression-like behavior by increasing hippocampal brain-derived neurotrophic factor levels in mice. Science Advances, 3(5), e1603014.
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5

Methylcellulose-based Colony Assay

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Cells were plated in methylcellulose-based medium (Methcoult H4230; Stemcell Technologies, Vancouver, BC, Canada) mixed with DMEM or RPMI-1640 (1:5). Thereafter, the cells were harvested, suspended in methylcellulose (1:10), poured into 24-well plates, and incubated for 5 days at 37°C in 5% CO2. Cell colonies were stained using p-iodonitrotetrazolium violet and visualized using the FluorChem 8800 imaging system (Alpha Innotech, San Leandro, CA).
Kaseb A.O., Haque A., Vishwamitra D., Hassan M.M., Xiao L., George B., Sahu V., Mohamed Y.I., Carmagnani Pestana R., Lombardo J.L., Avritscher R., Yao J.C., Wolff R.A., Rashid A., Morris J.S, & Amin H.M. (2022). Blockade of growth hormone receptor signaling by using pegvisomant: A functional therapeutic strategy in hepatocellular carcinoma. Frontiers in Oncology, 12, 986305.
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6

Bladder Collagen Protein Analysis

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The urinary bladder was freshly dissected out and homogenized in T-per buffer (Pierce Biotechnology, Rockford, IL) supplemented with protease and phosphatase inhibitor cocktails (Sigma). The protein extracts were subject to centrifugation at 20,200 g for 10 min at 4°C, and the supernatant was removed to a fresh tube. The protein concentration was determined using Bio-Rad DC protein assay kit. Proteins were then separated on a 10% SDS-PAGE gel and transferred to a nitrocellulose membrane. The membrane was blocked with 5% milk in Tris-buffered saline for 1 h and then incubated with a primary antibody against type I collagen (1∶1000, Cell Signaling Technology, Inc.) followed by IRDye secondary antibody. For internal loading control and normalization, the same membrane was striped and re-probed with anti-β-actin (1∶3000, Sigma). The bands were visualized with an ODYSSEY infrared imaging system (Li-cor Bioscience). Densitometric quantification of immunoreactive bands was performed using the software FluorChem 8800 (Alpha Innotech, San Leabdro, CA).
Qiao Z., Xia C., Shen S., Corwin F.D., Liu M., Guan R., Grider J.R, & Qiao L.Y. (2014). Suppression of the PI3K Pathway In Vivo Reduces Cystitis-Induced Bladder Hypertrophy and Restores Bladder Capacity Examined by Magnetic Resonance Imaging. PLoS ONE, 9(12), e114536.
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7

Western Blot Analysis of Protein Phosphorylation

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The protein extracts were subject to centrifugation at 20,200g for 10 min at 4 °C, and the supernatant was removed to a fresh tube. The protein concentration was determined using Bio-Rad DC protein assay kit. Proteins were then separated on a 10 % SDS-PAGE gel and transferred to a nitrocellulose membrane. The membrane was blocked with 5 % milk in Tris-buffered saline for 1 h and then incubated with a specific primary antibody followed by HRP-conjugated secondary antibody. For internal loading control, the same membrane was stripped and re-probed with anti-β-actin antiserum or antibody to a non-phosphorylated form of the protein examined. The concentrations of antibodies used were p-NR1: 1:1000; p-Akt and Akt: 1:1000; and β-actin: 1:3000. The bands were visualized by enhanced chemiluminescence (ECL). Densitometric quantification of the immunoreactive bands was performed using the software FluorChem 8800 (Alpha Innotech, San Leandro, CA).
Liu M., Kay J.C., Shen S, & Qiao L.Y. (2015). Endogenous BDNF augments NMDA receptor phosphorylation in the spinal cord via PLCγ, PKC, and PI3K/Akt pathways during colitis. Journal of Neuroinflammation, 12, 151.
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8

Colony Formation Assay for Hematopoietic Cells

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Cells were plated in a methylcellulose-based medium (Methocult H4230, Stemcell Technologies) and mixed in RPMI-1640 medium at a ratio of 1:4 (v/v) [67 (link), 68 (link)]. Harvested cells were mixed in a 1:10 (v/v) ratio with methylcellulose in 15 mL conical tubes that were then inverted gently. Thereafter, the contents were poured into 24-well plates, and incubated at 37°C in a 5% CO2 incubator for 5 days. Then, p-iodonitrotetrazolium violet was added and incubated overnight. Colonies were visualized using the FluorChem 8800 imaging system (Alpha Innotech, San Leandro, CA).
Yang L., Zhang S., George S.K., Teng R., You X., Xu M., Liu H., Sun X., Amin H.M, & Shi W. (2015). Targeting Notch1 and proteasome as an effective strategy to suppress T-cell lymphoproliferative neoplasms. Oncotarget, 6(17), 14953-14969.
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