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Anti nfia

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Anti-NFIA is a laboratory reagent that targets the Nuclear Factor I A (NFIA) protein. It is a tool used in research applications to study the role and function of the NFIA transcription factor in biological systems. The core function of Anti-NFIA is to provide a specific and selective means of detecting and quantifying NFIA expression or activity in experimental models.

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Lab products found in correlation

Anti h3k27ac, by Thermo Fisher Scientific (2 mentions) Nextseq 2000, by Illumina (2 mentions) Pag mnase, by Cell Signaling Technology (2 mentions) Anti nfia, by Thermo Fisher Scientific (2 mentions) Anti nfix, by Merck Group (2 mentions) Anti β actin, by Merck Group (1 mentions) Anti flag m2, by Merck Group (1 mentions) Hpa006111, by Abcam (1 mentions) Anti oxphos, by Abcam (1 mentions) Anti nf κb p65, by Cell Signaling Technology (1 mentions) Anti actin, by Santa Cruz Biotechnology (1 mentions) Flag m2, by Merck Group (1 mentions)

4 protocols using anti nfia

1

CUT&RUN Profiling of Chromatin Regulators

Cited 1 time
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0.5-1 million undifferentiated HUDEP2, HUDEP2 at day 2 of differentiation, or primary cells at day 8 of differentiation were used for CUT&RUN. We followed the experimental procedures published by Henikoff laboratory with minor adaptations35 . Antibody incubations were performed in 300 μl PCR tubes for 2 h at 4 °C with rotation. pAG-MNase were obtained from Cell Signaling. Antibodies used in this study were: IgG (Cell signaling, #3900), anti-NFIA (Thermo Fisher Scientific, PA5-52252), anti-NFIX (Sigma Aldrich, SAB1401263), anti-H3K27ac (Invitrogen, MA5-23516). anti-NFIA antibodies used in Extended Data Figure 6 were obtained from Sigma, #HPA008884; Active Motif, #39397; Invitrogen, #535936. Approximately 2 μg antibody were used per 200 μl reaction. The released DNA fragments were quantified by Qubit and sequencing libraries were generated using NEB Kits (E7645 and E6440S) with 15-30 ng input DNA6 (link). The barcoded libraries were pooled and paired-end (2×50 bp) sequenced on the Illumina Nextseq 2000 platform.
Sequencing data from CUT&RUN were analyzed by CUT-RUNTOOLS-2.064 (link) using default settings (https://github.com/fl-yu/CUT-RUNTools-2.0).
Qin K., Huang P., Feng R., Keller C.A., Peslak S.A., Khandros E., Saari M., Lan X., Mayuranathan T., Doerfler P.A., Abdulmalik O., Giardine B., Chou S.T., Shi J., Hardison R.C., Weiss M.J, & Blobel G.A. (2022). Dual function NFI factors control fetal hemoglobin silencing in adult erythroid cells. Nature genetics, 54(6), 874-884.
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2

Western Blot Analysis of Nuclear Transcription Factors

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Tissues were lysed in radioimmunoprecipitation assay (RIPA) buffer containing 0.1% SDS, 1% NP-40, 0.5% Na deoxycholate, 150 mM NaCl, 50 mM Tris-Cl (pH 8.0), 1 mM EDTA supplemented with protease inhibitor (Roche). Proteins were separated by SDS-PAGE, transferred to nitrocellulose membrane, and detected with the antibodies anti-NFIA (Sigma HPA006111), anti-FLAG M2 (Sigma F3165), KLF5 (Abcam ab137676) and anti-β actin (Sigma A3854).
Hiraike Y., Waki H., Miyake K., Wada T., Oguchi M., Saito K., Tsutsumi S., Aburatani H., Yamauchi T, & Kadowaki T. (2020). NFIA differentially controls adipogenic and myogenic gene program through distinct pathways to ensure brown and beige adipocyte differentiation. PLoS Genetics, 16(9), e1009044.
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3

Protein Extraction and Western Blot Analysis

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Tissues were lysed in RIPA buffer containing 0.1% SDS, 1% NP-40, 0.5% Na deoxycholate, 150 mM NaCl, 50 mMTris-Cl (pH 8.0), and 1 mMEDTA supplemented with protease inhibitor (Roche). Total proteins were loaded and fractionated by SDS-PAGE, transferred to a nitrocellulose membrane, and detected with the antibodies anti-NFIA (1:500 dilution, Sigma, HPA006111), anti-Ox-Phos (1:1,000 dilution, Abcam, 110413), anti-actin (1:1,000 dilution, Santa Cruz Biotechnology, sc-1616), anti-NF-κB p65 (1:1,000 dilution, Cell Signaling, 8242), and FLAG M2 (1:2,000 dilution, Sigma, F3165).
Hiraike Y., Saito K., Oguchi M., Wada T., Toda G., Tsutsumi S., Bando K., Sagawa J., Nagano G., Ohno H., Kubota N., Kubota T., Aburatani H., Kadowaki T., Waki H., Yanagimoto S, & Yamauchi T. (2023). NFIA in adipocytes reciprocally regulates mitochondrial and inflammatory gene program to improve glucose homeostasis. Proceedings of the National Academy of Sciences of the United States of America, 120(31), e2308750120.
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4

CUT&RUN Profiling of Chromatin Regulators

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
0.5-1 million undifferentiated HUDEP2, HUDEP2 at day 2 of differentiation, or primary cells at day 8 of differentiation were used for CUT&RUN. We followed the experimental procedures published by Henikoff laboratory with minor adaptations35 . Antibody incubations were performed in 300 μl PCR tubes for 2 h at 4 °C with rotation. pAG-MNase were obtained from Cell Signaling. Antibodies used in this study were: IgG (Cell signaling, #3900), anti-NFIA (Thermo Fisher Scientific, PA5-52252), anti-NFIX (Sigma Aldrich, SAB1401263), anti-H3K27ac (Invitrogen, MA5-23516). anti-NFIA antibodies used in Extended Data Figure 6 were obtained from Sigma, #HPA008884; Active Motif, #39397; Invitrogen, #535936. Approximately 2 μg antibody were used per 200 μl reaction. The released DNA fragments were quantified by Qubit and sequencing libraries were generated using NEB Kits (E7645 and E6440S) with 15-30 ng input DNA6 (link). The barcoded libraries were pooled and paired-end (2×50 bp) sequenced on the Illumina Nextseq 2000 platform.
Sequencing data from CUT&RUN were analyzed by CUT-RUNTOOLS-2.064 (link) using default settings (https://github.com/fl-yu/CUT-RUNTools-2.0).
Qin K., Huang P., Feng R., Keller C.A., Peslak S.A., Khandros E., Saari M., Lan X., Mayuranathan T., Doerfler P.A., Abdulmalik O., Giardine B., Chou S.T., Shi J., Hardison R.C., Weiss M.J, & Blobel G.A. (2022). Dual function NFI factors control fetal hemoglobin silencing in adult erythroid cells. Nature genetics, 54(6), 874-884.
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