After culture, cells were re-sorted (97-99% purity) and cross-linked with 1% formaldehyde for use in ChIP experiments. Anti-Foxp3 (Abcam, ab2481) ChIPs was performed with an A/G agarose ChIP kit (Thermo Scientific), following the manufacturers protocol. After enzymatic digestion, 10% of the chromatin was saved as input controls. The remaining chromatin was immunoprecipitated with 1 μg of antibody during an overnight incubation at 4°C. After IP elution, cross-linking was performed on the purified product and DNA fragments collected for use in real-time qPCR following the described cycling conditions and with primers listed in Table I . The relative enrichment was calculated using input controls and a delta-delta Ct equation as follows: (2^-(CtIL2, IP – CtGapdh, IP))/(2^-(CtIL2, input-CtGapdh, input))
Ab2481
Manufactured by Abcam
Sourced in United Kingdom
Ab2481 is a laboratory equipment product. It is a high-performance device used for various applications in scientific research and analysis. The core function of this product is to provide reliable and accurate measurements or data processing capabilities to support research activities.
Automatically generated - may contain errors
Lab products found in correlation
Confocal microscopy, by Zeiss (1 mentions)
Image pro plus software, by Media Cybernetics (1 mentions)
Alexa fluor 488 conjugated anti mouse igg, by Abcam (1 mentions)
Alexa fluor 647 conjugated anti rabbit igg, by Abcam (1 mentions)
Ab150117, by Abcam (1 mentions)
Ab9657, by Abcam (1 mentions)
Ab34843, by Abcam (1 mentions)
Ab150075, by Abcam (1 mentions)
Alexa fluor 555 conjugated anti rabbit igg, by Abcam (1 mentions)
Ab150074, by Abcam (1 mentions)
Ab2302, by Abcam (1 mentions)
2 protocols using ab2481
1
Foxp3 ChIP-qPCR Protocol for Regulatory T Cells
2
Immunohistochemical Analysis of HCC Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Paraffin-embedded and formalin-fixed HCC samples were used for immunohistochemistry detection as previously described55 (link). Antibodies (Ab) including human EGFR (1:100, ab30), Foxp3 (1:100, ab2481), IFN-γ (1:200, ab9657), cleaved Caspase 3 (C.C3) (1:200, ab2302) and IL-10 (1:100, ab34843) (Abcam, Cambridge, UK) were used. For immunofluorescence analysis, tissue slides were stained with rabbit anti-human Foxp3, rabbit anti-human cleaved Caspase 3, mouse anti-human EGFR, and goat anti-human IFN-γ antibodies, followed by staining with Alexa Fluor 488-conjugated anti-mouse IgG (1:500, Ab150117), Alexa Fluor 555-conjugated anti-rabbit IgG(1:1,000, Ab150074), and Alexa Fluor 647-conjugated anti-Rabbit IgG (1:500, Ab150075) (Abcam, Cambridge, UK) antibodies. Positive cells were quantified using Image-Pro Plus software (Media Cybernetics, MD, USA) and detected by confocal microscopy (Zeiss, Oberkochen, Germany).
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