Am4300

Manufactured by Abcam

Sourced in United States

The AM4300 is a microarray scanner designed for high-throughput analysis of DNA and protein samples. It features a sensitive photodetector and advanced optics to deliver accurate and reproducible results. The scanner supports a range of slide formats and can process multiple samples simultaneously.

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Lab products found in correlation

7 protocols using am4300

Western Blot Protein Quantification

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Cell pellets were lysed with RIPA buffer for 30 min on ice. After centrifugation at 12,000 g for 30 min at 4°C, the supernatants were collected. The protein concentrations were measured with a biophotometer (Eppendorf). 20 μg total proteins of each sample were applied for SDS-PAGE electrophoresis; then the gel was transferred to PVDF membranes. The following primary antibodies were used: anti-GAPDH (Ambion, AM4300), anti-GluA1 (Abcam, ab1232), anti-Flag (Thermo, MA1-91878), and anti-HA (Diagbio, db5297). The images were measured by Molecular Imager Imaging System (Tanon, China). The intensity of images was analyzed with Adobe Photoshop software.

Xu W., Zhang X., Liang F., Cao Y., Li Z., Qu W., Zhang J., Bi Y., Sun C., Zhang J., Sun B., Shu Q, & Li X. (2021). Tet1 Regulates Astrocyte Development and Cognition of Mice Through Modulating GluA1. Frontiers in Cell and Developmental Biology, 9, 644375.

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Antiviral Effects of SP-2509 and OG-L002 on HSV-1 Infection

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MRC-5 cells were seeded at a density of 1 × 10⁶ cells/well in a 6-well dish and infected with strain KOS at an MOI of 10 PFU/cell. Treated cells were incubated with 16 μM SP-2509 or 60 μM OG-L002 for 1 h prior to and during infection. At the indicated times postinfection, proteins were isolated from cells using Laemmli SDS sample buffer and Western blotting was carried out using the following primary antibodies: α-ICP4 (58S), α-ICP27 (P1113), α-ICP8 (Abcam ab20194), α-UL42 (Abcam ab19311), α-VP5/ICP5 (Abcam ab6508), α-gC (GICR 1104), α-GAPDH (Thermo AM4300), and α-LSD1 (Abcam ab37165). The intensity of gel bands was quantified using the GelAnalyzer plugin in ImageJ. Band intensities were normalized to GAPDH detected from the same sample.

Harancher M.R., Packard J.E., Cowan S.P., DeLuca N.A, & Dembowski J.A. (2020). Antiviral Properties of the LSD1 Inhibitor SP-2509. Journal of Virology, 94(19), e00974-20.

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Antibody Characterization for EBOV Studies

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The antibodies used in this study include monoclonal anti-polyhistidine−alkaline phosphatase antibody (Milliposre Sigma #A5588, St. Louis, MO, USA), anti-EGFP monoclonal antibody (ThermoFisher Scientific #MA1-952, Grand Island, NY, USA), anti-HA tag antibody (Abcam #ab18181, Cambridge, MA, USA), anti-EBOV VP24 polycolonal antibody (IBT Bioserveices #0301-047, Rockville, MD, USA), anti-EBOV VP30 polycolonal antibody (IBT Bioserveices #0301-048, Rockville, MD, USA), anti-EBOV VP40 polycolonal antibody (IBT Bioserveices #0301-010, Rockville, MD, USA), anti-GAPDH monoclonal antibody (6C5) (ThermoFisher Scientific #AM4300, Grand Island, NY, USA), anti-alpha 1 sodium potassium ATPase antibody (Abcam #ab7671, Cambridge, MA, USA), anti-LAMP2 antibody (Abcam #ab13524, Cambridge, MA, USA), and anti-histone H3 antibody (Abcam #ab24834, Cambridge, MA, USA).

Su Y, & Stahelin R.V. (2020). The Minor Matrix Protein VP24 from Ebola Virus Lacks Direct Lipid-Binding Properties. Viruses, 12(8), 869.

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Western Blot Antibody Optimization

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The antibody target, dilution, species, company and product number used for Western blot analysis are as follows: Cirbp 1:500 (rabbit, Proteintech, 10209–2-AP); GAPDH 1:2000 (mouse, Invitrogen, AM4300); Vinculin 1:1000 (mouse, Abcam, ab130007). Secondary antibodies were goat anti-rabbit (LI-COR IRDye 800CW, 926–32211) and goat anti-mouse (LI-COR IRDye 680RD, 926–68070).

King B.M., Mintz S., Lin X., Morley G.E., Schlamp F., Khodadadi-Jamayran A, & Fishman G.I. (2023). Chronic Kidney Disease Induces Pro-Arrhythmic Remodeling. Circulation. Arrhythmia and electrophysiology, 16(1), e011466.

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Comprehensive Protein Isolation and Analysis

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Protein isolation was performed using working RIPA buffer (120mM NaCl, 50 mM pH 8.0 Tris, 0.5% NP-40, 1 mM EGTA) containing protease and phosphatase inhibitors (100 ug/mL PMSF, 1 mM NaOrVa, 50 ug/mL aprotinin, 50ug/mL leupeptin). Protein concentration was determined using a Bio-Rad protein concentration assay dye and absorbance read at 595 nm on a spectrophotometer (SpectraMax ID3, Molecular Devices). 20μg total protein in βmercaptoethanol-containing loading buffer was added per well onto 4–12% SDS-PAGE gels (NuPAGE) and transferred to PVDF (Bio-Rad) or nitrocellulose (Amersham) membrane. nitrocellulose membranes were stained for total protein using Ponceau S (Boston BioProducts). Membranes were blocked with 10% BSA or milk-fat in TBST for 1 hour at room temperature. Primary antibodies from Abcam for AR (ab74272), AR-V7 (ab198394), phosphorylated AR-Ser650 (ab47563), ACAT1 (ab168342), MAP3K11 (ab51068), PSDM12 (ab229930), and GAPDH (AM4300) were used at recommended concentrations in 2.5% milk-fat or 2.5% bovine serum albumin overnight at 4°C. Anti-rabbit and anti-mouse secondary antibodies (GE Healthcare) were used in 2.5% milk-fat for one hour at room temperature. Membranes were imaged via chemiluminescence reagents (Super Signal™ West Pico Plus, Thermo Fisher) on a digital imager (ChemiDoc™ Touch, Bio-Rad).

Kohrt S.E., Awadallah W.N., Phillips RA I.I.I., Case T.C., Jin R., Nanda J.S., Yu X., Clark P.E., Yi Y., Matusik R.J., Anderson P.D, & Grabowska M.M. (2020). Identification of genes required for enzalutamide resistance in castration-resistant prostate cancer cells in vitro. Molecular cancer therapeutics, 20(2), 398-409.

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Western Blot Protein Analysis

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Samples were transferred to nitrocellulose (PVDF for Fubi blots) membranes using a Trans-Blot Turbo system (Bio-Rad). Membranes were blocked with 5% (wt/vol) nonfat milk in PBS-T (PBS and 0.1% Tween 20) and incubated overnight with the indicated primary antibodies (anti-FAU, 1:1,000, Proteintech, 13581-1-AP; anti-Flag M2, 1:2,000, Sigma, F3165; anti-HA, 1:1,000, BioLegend, 16B12; anti-GAPDH, 1:10,000, Thermo Fisher, AM4300; USP7, 1:2,000, Abcam, ab190183; USP16, 1:1,000, Biomol, A301-614A-T; USP36, 1:500, Biomol, A300-940A-T). The signal was developed with secondary antibodies coupled to horseradish peroxidase (anti-mouse, 1:5,000, Sigma, NXA931; anti-rabbit, 1:5,000, Sigma, GENA934) and Clarity Western ECL substrate (Bio-Rad) supplemented with Clarity Max Western ECL substrate (Bio-Rad) when necessary. Chemiluminescence was imaged on a Chemi-Doc MP Imaging System (Bio-Rad), and quantification was performed using Image Lab (Bio-Rad).

O’Dea R., Kazi N., Hoffmann-Benito A., Zhao Z., Recknagel S., Wendrich K., Janning P, & Gersch M. (2023). Molecular basis for ubiquitin/Fubi cross-reactivity in USP16 and USP36. Nature Chemical Biology, 19(11), 1394-1405.

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Western Blot Analysis of Lipid Metabolism Proteins

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Blots were blocked for 30 min in 2.5% (m/v) bovine serum albumin (BSA) in Tris-buffered saline with Tween-20 (TBST) (10 mM Tris, 0.15 M NaCl, pH 8, 0.05% Tween-20). Blots were incubated with primary antibodies PAFAH1B3 1:200 (Santa Cruz; sc-393612), PAFAH1B1 1:500 (Abcam; ab2607), LIPC 1:500 (Santa Cruz; sc-21740), ACOT1/2 1:500 (Santa Cruz; sc-373917), GAPDH 1:10,000 (Ambion; AM4300), β-tubulin 1:4000 (Abcam; ab6046), and PPARG 1:500 (Abcam; ab178860) in 2.5% BSA in TBST overnight at 4 °C. Blots were incubated for 1 h at room temperature (RT) with the appropriate secondary antibody, peroxidase AffiniPure goat antimouse immunoglobulin G (H + L) (Jackson ImmunoResearch Laboratories; 115-035-062) or peroxidase AffiniPure Donkey anti-rabbit immunoglobulin G (H + L) (Jackson ImmunoResearch Laboratories; 711-035-152), at 1:20,000 in 2.5% BSA in TBST. Densitometric analysis was performed using ImageLab 6.0 software (Bio-Rad) and significance was calculated using two-tailed paired t tests.

Desrochers G.F., Filip R., Bastianelli M., Stern T, & Pezacki J.P. (2022). microRNA-27b regulates hepatic lipase enzyme LIPC and reduces triglyceride degradation during hepatitis C virus infection. The Journal of Biological Chemistry, 298(6), 101983.

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