Anti ahr antibody

Manufactured by Thermo Fisher Scientific

Sourced in United Kingdom, United States

The Anti-AhR antibody is a laboratory reagent used to detect and quantify the Aryl Hydrocarbon Receptor (AhR) protein in various biological samples. AhR is a transcription factor that plays a crucial role in regulating the expression of genes involved in xenobiotic metabolism, cell proliferation, and other cellular processes. This antibody can be used in techniques such as Western blotting, immunoprecipitation, and immunohistochemistry to study the expression and localization of AhR in different cell types and tissues.

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Lab products found in correlation

Protein a g immunoprecipitation magnetic beads, by Selleck Chemicals (1 mentions) Mg132, by AbMole (1 mentions) Bz x810, by Keyence (1 mentions) Anti loricrin antibody, by Abcam (1 mentions)

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Anti-AHR (8 citations)

4 protocols using anti ahr antibody

Detecting Ubiquitinated AhR in Trophoblasts

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Trophoblast cells were treated with MG132 (M1902, AbMole Bioscience, USA) for 6 h, and then lyzed, in lysis buffer containing protease inhibitors. Cell lysates were incubated with 30 µL Protein A/G immunoprecipitation magnetic beads (B23201, Bimake), coupling with anti‐AhR antibody (1:250, PA5‐122264, Invitrogen) or anti‐IgG antibody at 4 °C overnight. Subsequently, the ubiquitinated AhR (AhR‐Ub) was detected by Western blotting.

Chen W., Mi C., Zhang Y., Yang Y., Huang W., Xu Z., Zhao J., Wang R., Wang M., Wan S., Wang X, & Zhang H. (2024). Defective Homologous Recombination Repair By Up‐Regulating Lnc‐HZ10/Ahr Loop in Human Trophoblast Cells Induced Miscarriage. Advanced Science, 11(13), 2207435.

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Immunofluorescence analysis of AKR1C3, loricrin, and AHR

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Tissues were obtained by biopsy from patients and fixed in 10% formalin before embedding in paraffin. Sections were deparaffinized in xylene for 10 min, followed by graded rehydration in EtOH (99.5%, 95%, 85%, 70%, and 50%) for 5 min each. For AKR1C3 and loricrin, sections were incubated in antigen retrieval buffer (10 mM Tris, 1 mM EDTA, 0.05% Tween-20, pH 6.0) at 95 °C for 10 min in a water bath. For AHR, sections were incubated in antigen retrieval buffer (10 mM Tris, 1 mM EDTA, pH 9.0) at 95 °C for 10 min in a pressure cooker. Sections were incubated in the following primary antibodies, 1:46 anti-AKR1C3 antibody (Sigma-Aldrich, Darmstadt, Germany), 1:500 anti-loricrin antibody (Abcam, Cambridge, UK), or 1:50 anti-AHR antibody (Invitrogen, Waltham, MA) overnight at 4 °C. Sections were then incubated in 1:500 Alexa Fluorescein 594-conjugated goat anti-mouse IgG antibody (Invitrogen), 1:500 Alexa Fluorescein 488-conjugated secondary goat anti-rabbit IgG antibody (Invitrogen) for 30 min at 37 °C. The images were obtained using a fluorescence microscope BZ-X810 (Keyence, Osaka, Japan). The fluorescence intensities of AKR1C3 and AHR were calculated using ImageJ software (NIH, Bethesda, MD) from 10 randomly selected fields.

Nojiri Y., Nakamura M., Magara T., Yamamoto A., Ikumi K., Nakamura R., Nishida E., Haarmann-Stemmann T, & Morita A. (2023). Influence of aldo–keto reductase 1C3 polymorphisms in early-onset female psoriasis patients. Scientific Reports, 13, 3280.

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Chromatin Immunoprecipitation (ChIP) Assay for AHR Binding

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Chromatin immunoprecipitation (ChIP) assays were performed according to the kit protocol (Santa Cruz Biotechnology, Visalia, CA) with an anti-AHR antibody (Thermo Scientific, Rockland, IL). The PCR product corresponding to the Ubch7 proximal gene promoter was generated from an aliquot of immunoprecipitated material. Brain homogenates from WT mice treated with a single oral dose of TCDD or vehicle (corn oil) were washed with PBS buffer and cross-linked with 1% formaldehyde. After chromatin isolation, the DNA was fragmented, and immunoprecipitation was performed. The cross-linking was reversed, the DNA was purified, and PCR amplification was performed as follows: initial denaturation at 94^◦C for 3 min, 25 cycles of denaturation at 94^◦C for 30 s, annealing at 65^◦C for 30 s, and extension at 72^◦C for 3 s. A final extension cycle at 72^◦C for 10 min was added to the end of the program. The oligonucleotides used for PCR amplification were 5^′-GGCTAGAACCCCCTCACTTC-3^′ (forward) and 5´-GGCTAGAACCCCCTCACTTC-3 (reverse).

González-Barbosa E., Mejía-García A., Bautista E., Gonzalez F.J., Segovia J, & Elizondo G. (2017). TCDD induces UbcH7 expression and synphilin-1 protein degradation in the mouse ventral midbrain. Journal of biochemical and molecular toxicology, 31(10), 10.1002/jbt.21947.

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ChIP Assay for AhR-Binding to Prkn Promoter

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ChIP assays were performed according to the kit protocol (Santa Cruz Biotechnology, Visalia, CA, USA) with an anti-AhR antibody (Thermo Fisher Scientific, Cat. MA1–514, Rockland, IL, USA). The PCR product corresponding to the Prkn proximal gene promoter was generated from an aliquot of immunoprecipitated material. Brain homogenates from WT mice treated with a single oral dose of TCDD or vehicle (corn oil) were washed with PBS buffer and crosslinked with 1% formaldehyde. After chromatin isolation, the DNA was fragmented, and immunoprecipitation was performed. The crosslinking was reversed, the DNA was purified, and PCR amplification was performed as follows: initial denaturation at 94 °C for 3 min and 28 cycles of denaturation at 94 °C for 30 s, annealing at 65 °C for 30 s, and extension at 72 °C for 30 s. A final extension cycle at 72 °C for 10 min was added to the end of the program. The oligonucleotides used for PCR amplification were 5′-AGAAGTGAGCAGGGGGTCGGG-3′ (forward) and 5′-AAGGACCTAC GCGGGCACTG-3′ (reverse).

González-Barbosa E., García-Aguilar R., Vega L., Cabañas-Cortés M.A., Gonzalez F.J., Segovia J., Morales-Lázaro S.L., Cisneros B, & Elizondo G. (2019). Parkin is transcriptionally regulated by the aryl hydrocarbon receptor: Impact on α-synuclein protein levels. Biochemical pharmacology, 168, 429-437.

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