Anti batf

Manufactured by Cell Signaling Technology

Sourced in United States

Anti-BATF is a primary antibody product offered by Cell Signaling Technology. It is designed to detect the BATF (Basic Leucine Zipper ATF-Like Transcription Factor) protein. BATF is a transcription factor involved in the regulation of gene expression.

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Anti-BATF (D7C5) (3 citations) Anti-mouse BATF (D7C5) (2 citations)

5 protocols using anti batf

Chromatin Immunoprecipitation Antibody Panel

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anti-H3K27ac (Cat# 8173), anti-Batf (Cat# 8638), anti-H3K27me3 (Cat# 9733), anti-Mbd3 (Cat# 99169), anti-Chd3 (Cat# 4241), anti-Rbap46 (Cat# 6882), anti-Hdac1 (Cat# 34589), anti-Hdac2 (Cat# 57156), anti-EEA1 (Cat# 3288), anti-H3 (Cat# 4499), and anti-β-actin (Cat# 3700) were from Cell Signaling Technology (Danvers, USA). Anti-Kcnt2 (Cat# bs-12177R) was from Bioss Antibodies (Beijing, China). Anti-CD127 (A7R34), anti-c-Kit (2B8), anti-CD3 (17A2), anti-CD4 (GK1.5), anti-CD19 (1D3), anti-NK1.1 (PK136), anti-CD150 (mShad150), anti-CD34 (RAM34), anti-CD45 (30-F11), anti-CD90 (HIS51), anti-Sca-1 (D7), anti-CD25 (PC61.5), anti-Flt3 (A2F10), anti-α₄β₇ (DATK32), anti-RORγt (AFKJS-9), anti-NKp46 (29A1.4), anti-Gata3 (TWAJ), anti-KLRG1 (2F1), anti-PLZF (Mags.21F7), Lineage cocktail (88-7772-72), anti-CD48 (HM48-1), Anti-IL-17 (eBio17B7), and anti-CD16/32 (93) were purchased from eBiosciences (San Diego, USA). Anti-BrdU (600-401-C29) was purchased from ThermoFisher. Paraformaldehyde (PFA) and 4′,6-diamidino-2-phenylindole (DAPI) were from Sigma-Aldrich. The IL-17 ELISA kit was purchased from eBiosciences.

Liu B., Ye B., Zhu X., Yang L., Li H., Liu N., Zhu P., Lu T., He L., Tian Y, & Fan Z. (2020). An inducible circular RNA circKcnt2 inhibits ILC3 activation to facilitate colitis resolution. Nature Communications, 11, 4076.

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Immunoblot Analysis of IRF4, BATF, and β-actin

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Protein extracts were resolved by SDS-PAGE, transferred onto an Immunobilon membrane, and analyzed by immunoblot with anti-IRF4 (sc-6059; Santa Cruz), anti-BATF (8638; Cell Signaling Technology), and anti-β-actin (12262; Cell Signaling Technology). Horseradish peroxidase–linked antibody to rabbit immunoglobulin G (7074; Cell Signaling Technology), horseradish peroxidase–linked antibody to goat immunoglobulin G (sc-2768; Santa Cruz), and horseradish peroxidase–linked antibody to mouse immunoglobulin G (7076; Cell Signaling Technology) were used as secondary antibodies. Protein expression was detected by chemiluminescence.

Wu J., Zhang H., Shi X., Xiao X., Fan Y., Minze L.J., Wang J., Ghobrial R.M., Xia J., Sciammas R., Li X.C, & Chen W. (2017). Ablation of transcription factor IRF4 promotes transplant acceptance by driving allogenic CD4+ T cell dysfunction. Immunity, 47(6), 1114-1128.e6.

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Immunoblot Analysis of Protein Signaling

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Protein extracts were resolved by SDS–PAGE, transferred onto an Immunobilon membrane, and analysed by immunoblot with the following specific antibodies: anti-p105/p50 (ab32360, 1:5,000), anti-NF-κB p65 (RelA) (ab16502, 1:5,000), anti-p300 (ab3164, 1:1,000), anti-HDAC2 (ab7029, 1:1,000), anti-Sirt1 (ab110304, 1:1,000; all from Abcam); anti-p100/p52 (4882, 1:1,000), anti-RelB (4922, 1:1,000), anti-Histone H3 (4499, 1:2,000), anti-β-actin (12262, 1:2,000), anti-Batf (8638, 1:1,000), anti-PU.1 (2258, 1:1,000), anti-Phospho-STAT5 (9359, 1:1,000), anti-Smad2 (5339, 1:1,000), anti-Phospho-Smad2 (3108, 1:1,000), anti-Smad3 (9523, 1:1,000), anti-Phospho-Smad3 (9520, 1:1,000), anti-Sirt7 (5360, 1:1,000), anti-HDAC1 (2062, 1:1,000; all from Cell Signaling Technology); and anti-STAT5 (SC-835X, 1:3,000), anti-STAT6 (SC-981X, 1:4,000), anti-phospho-STAT6 (SC-11762X, 1:3,000), anti-IRF4 (sc-6059, 1:1,000; all from Santa Cruz Technology). Horseradish peroxidase (HRP)-linked antibody to mouse IgG (7076, 1:2,000), HRP–linked antibody to rabbit IgG (7074, 1:2,000; both from Cell Signaling Technology) and HRP-linked antibody to goat IgG (sc-2768, 1:2,000; Santa Cruz Technology) were used as secondary antibodies. The expression of target molecules was detected by chemoluminescence method. Images has been cropped for presentation; full-size immunoblots are provided in Supplementary Fig. 7.

Xiao X., Shi X., Fan Y., Zhang X., Wu M., Lan P., Minze L., Fu Y.X., Ghobrial R.M., Liu W, & Li X.C. (2015). GITR subverts Foxp3+ Tregs to boost Th9 immunity through regulation of histone acetylation. Nature Communications, 6, 8266.

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Immunoblotting and ELISA for T-cell Analysis

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Immunoblotting was performed as previously described^{31 (link)} using anti-IRF4 (Santa Cruz, Dallas, TX, USA), anti-BATF (Cell Signaling, Beverly, MA, USA), and anti-β-Actin antibodies (Cell Signaling). IL-2 enzyme-linked immunosorbent assay (ELISA) was performed using cell-free supernatants from conditioned T-cell medium and a mouse IL-2 ELISA Ready-SET-Go! Kit following the manufacturer's instructions (eBioscience).

Grusdat M., McIlwain D.R., Xu H.C., Pozdeev V.I., Knievel J., Crome S.Q., Robert-Tissot C., Dress R.J., Pandyra A.A., Speiser D.E., Lang E., Maney S.K., Elford A.R., Hamilton S.R., Scheu S., Pfeffer K., Bode J., Mittrücker H.W., Lohoff M., Huber M., Häussinger D., Ohashi P.S., Mak T.W., Lang K.S, & Lang P.A. (2014). IRF4 and BATF are critical for CD8+ T-cell function following infection with LCMV. Cell Death and Differentiation, 21(7), 1050-1060.

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ChIP-seq Analysis of Th17 Cell Transcription Factors

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Cells were polarized in Tr1 conditions for 3 days, fixed with 1% formaldehyde for 10 mins and quenched with 0.125M glycine. Chromatin fraction preparation and chromatin IP was performed using SimpleChIP Enzymatic Chromatin IP (Cell Signaling Technology) according to the manufacturer’s instructions. The following antibodies were used: anti-IRF1 (Santa Cruz sc-640X), anti-BATF (Cell Signaling 8638S), anti-c-Maf (Santa Cruz sc-7866), anti-AhR (Enzo Life Sciences BML-SA210-0025), anti-trimethyl-Histone H3 (Lys4) (Millipore 07-473), anti-trimethyl-Histone H3 (Lys27) (Millipore 07-449), anti-Histone H3 (acetyl K9 (Abcam ab4441). For Re-ChIP, chromatin fraction was prepared as described above, and the chromatin IP was prepared using Re-ChIP-IT kit (Active Motif) according to manufacturer’s instructions. ChIP and Re-ChIP quantitative PCR (qPCR) was run on Viia7 Real-Time PCR System (Applied Biosystems) and relative expression was calculated using Ct values of the samples and inputs. Primers spanning Il17a and IL23r sites were described before^{29 (link)}. The following primer pairs were used for the rest of the targets:

Karwacz K., Miraldi E.R., Pokrovskii M., Madi A., Yosef N., Wortman I., Chen X., Watters A., Carriero N., Awashti A., Regev A., Bonneau R., Littman D, & Kuchroo V.K. (2017). Critical role of the transcription factors IRF1 and BATF in preparing the chromatin landscape during Type 1 regulatory cell differentiation. Nature immunology, 18(4), 412-421.

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