Anti-KDM6B / JMJD3 antibody

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-KDM6B / JMJD3 antibody (AB169197)

ab169197 staining KDM6B/JMJD3 in MCF7 cells. The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab169197 at 1μg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control, at 1/1000 dilution. Cells were then incubated with ab150120, Goat polyclonal Secondary Antibody to Mouse at 1/1000 dilution (shown in pseudocolour red) and ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG at 1/1000 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

• WB

Lab

Western blot - Anti-KDM6B / JMJD3 antibody (AB169197)

Due to the cellular localisation of KDM6B / JMJD3 an increase in expression is expected within the Nuclear fraction.

This blot was produced using a 3-8% Tris Acetate gel under the TA buffer system. The gel was run at 150V for 60 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab169197 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.

All lanes:

Western blot - Anti-KDM6B / JMJD3 antibody (ab169197) at 1 µg/mL

Lane 1:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 2:

HeLa (Human epithelial carcinoma cell line) Nuclear Lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/10000 dilution

Predicted band size: 177 kDa

Observed band size: 177 kDa

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Exposure time: 4min

• WB

CiteAb

Western blot - Anti-KDM6B / JMJD3 antibody (AB169197)

KDM6B / JMJD3 western blot using anti-KDM6B / JMJD3 antibody ab169197. Publication image and figure legend from Solier, S., Müller, S., et al., 2023, Nature, PubMed 37100912.

ab169197 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab169197 please see the product overview.

Pharmacological inactivation of mitochondrial copper(II) attenuates inflammation in vivo.a, Western blots of copper-signalling effectors in SPMs from mice treated with LPS. Macrophages of several mice were pooled (4–7 mice per condition). b, Western blots of copper-signalling effectors in SPMs from mice subjected to CLP. Macrophages of several mice were pooled (7–8 mice per condition). H3 is a sample processing control. c, Western blots of copper-signalling effectors in AMs from K18-hACE2 mice infected with SARS-CoV-2. Macrophages of several mice were pooled (10 mice per condition). H3 is a sample processing control. d, Average body temperature of mice treated as indicated (n = 6–9 mice per group). e, GO term analysis of downregulated genes in lung tissues of SARS-CoV-2 infected K18-hACE2 mice treated with LCC-12 (0.5 mg/kg). f, RNA-seq analysis of gene expression in lung tissues of SARS-CoV-2-infected K18-hACE2 mice treated with LCC-12 (0.5 mg/kg) (n = 8 mice per group). Inflammatory signature genes highlighted. Dashed lines, adjusted P value = 0.05. g, Illustration of copper-signalling. Cell plasticity involves upregulation of the cell surface marker CD44, which mediates endocytosis of metal-bound hyaluronates. In the presence of copper(II), NADH reacts with H2O2 to replenish NAD+ in mitochondria, an enzyme cofactor involved in the biosynthesis of αKG and acetyl-CoA. These co-substrates of iron-dependent demethylases and acetyl-transferases are required for epigenetic and transcriptional programming of inflammation and the regulation of cell plasticity. Pharmacological inactivation of mitochondrial copper(II) blocks NAD(H) redox cycling, leading to distinct epigenetic states and transcriptional profiles. Targeting copper(II) interferes with cell plasticity in immune and cancer cells. For a – c gating strategy of SPMs and AMs see Methods and Supplementary Information. For d 2-way ANOVA. Mean values ± s.e.m. For e and f differential gene expression was assessed with the limma/voom framework. GO enrichment was assessed with the enrichGO method from clusterProfiler. P-values were corrected for multiple testing with the Benjamini-Hochberg procedure.Source data

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