ChIP assays were performed with the EZ-Magna ChipTM A kit (Millipore) using the manufacturer's instructions. A549 cells were treated with 2.5 μM TSA or 5 mM VPA for 24 h and cross-linked with 1% formaldehyde at room temperature for 10 min. The chromatin of A549 cells was sonicated on ice and then immunoprecipitated with anti-IgG antibody (Millipore), anti-GATA-1 antibody (Abcam), anti-Sp1 antibody (Santa Cruz), and anti-p300 antibody (Santa Cruz). After reverse cross-linking and DNA purification, DNA from input (1:10 diluted) or immunoprecipitated samples were assayed by RT-PCR. The 1% input and immunoprecipitated DNA were quantitatively compared by real time PCR using SYBR Green (TaKaRa). The IRF-3 promoter region, nt -149 to +18, containing GATA-1 element was amplified. Primer pairs for this region were: 5’-GTCTCCTCCACTGAACTCGTAC-3’ (sense) and 5’-GCCCTTTTTTGGGTTTCC-3’ (anti-sense).
Anti gata 1 antibody
Manufactured by Abcam
The Anti-GATA-1 antibody is a laboratory reagent used for the detection and quantification of the GATA-1 protein in biological samples. GATA-1 is a transcription factor that plays a crucial role in the development and differentiation of red blood cells and megakaryocytes. This antibody can be used in various applications, such as Western blotting, immunoprecipitation, and immunohistochemistry, to study the expression and localization of GATA-1 in different cell types and tissues.
Automatically generated - may contain errors
Lab products found in correlation
Ez magna chip tma kit, by Merck Group (1 mentions)
Anti p300 antibody, by Santa Cruz Biotechnology (1 mentions)
Sybr green, by Takara Bio (1 mentions)
Anti sp1 antibody, by Santa Cruz Biotechnology (1 mentions)
Ez magna chip a g kit, by Merck Group (1 mentions)
Nitrocellulose membrane, by Merck Group (1 mentions)
Anti gapdh antibody, by Bioworld Technology (1 mentions)
Protease inhibitor cocktail, by Keygen Biotech (1 mentions)
Bca protein assay kit, by Beyotime (1 mentions)
3 protocols using anti gata 1 antibody
1
GATA-1 and Sp1 Binding in IRF-3 Promoter
2
ChIP-qPCR Analysis of Hematopoietic Regulators
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For ChIP-qPCR assays, cells differentiated from cord blood CD34+ cells on days 4, 8 and 11, int1Δ6 K562 cells, int8Δ K562 cells, WT K562 cells and yolk sac cells from E11.5 mice were fixed with 1% (vol/vol) formaldehyde for 10 min at room temperature. The ChIP assays were performed using a EZ-Magna ChIP A/G Kit (Millipore, 17–408). An anti-GATA1 antibody (Abcam, ab11852), anti-TAL1 antibody (Santa Cruz, sc12984), anti-LDB1 antibody (Santa Cruz, sc11198), anti-LMO2 (Abcam, ab72841), anti-FOG1 antibody (Santa Cruz, sc9361), anti-Pol II antibody (Abcam, ab817) and normal mouse and rabbit IgGs were used for immunoprecipitation. ChIP DNA fragments, which ranged from 100 to 500 bp according to agarose gel electrophoresis, were subjected to real-time PCR analysis using primers that targeted the promoter region, intron 1 enhancer region or intron 8 enhancer region of the human ALAS2 gene. The primers used for the ChIP-qPCR analysis are listed in Supplementary Table S1.
3
Protein Expression Analysis of TSA and VPA
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Cells were treated with TSA or VPA at different concentration for 48 h. Total protein was extracted from cells and LADC tissue with lysis buffer containing protease inhibitor cocktail (Keygentec, China) and the concernrations were determined by the BCA Protein Assay Kit (Beyotime). Equal amount of protein were separated by sodiumdodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to a nitrocellulose membrane (Millipore). The membrane were blocked with 5% nonfat milk in TBST (0.1% Tween-20) and then incubated with anti-IRF-3 antibody (Abcam), anti-GATA-1 antibody (Abcam), or anti-GAPDH antibody (Bioworld) at 4 °C overnight at a dilution of 1:2000-1:4000, followed by anti-rabbit IgG (Cell signaling) or anti-mouse IgG (Jackson ImmunoResearch Laboratories) at a dilution of 1:5000. Intensity of bands was measured using an ECL imager.
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