Anti-KDM5C / Jarid1C / SMCX antibody (ab34718)
Key features and details
- Rabbit polyclonal to KDM5C / Jarid1C / SMCX
- Suitable for: WB, ICC/IF
- Knockout validated
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-KDM5C / Jarid1C / SMCX antibody
See all KDM5C / Jarid1C / SMCX primary antibodies -
Description
Rabbit polyclonal to KDM5C / Jarid1C / SMCX -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species ICC/IF HumanWB Human
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Immunogen
Synthetic peptide conjugated to KLH derived from within residues 1500 to the C-terminus of Human Jarid1C/ SMCX.
Read Abcam's proprietary immunogen policy (Peptide available as ab35501.) -
Positive control
- WB: HEK-293T, HAP1 and Y79 cell lysates. ICC: HeLa cells.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help. -
Concentration information loading... -
Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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ChIP Related Products
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab34718 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
| Application | Species |
|---|---|
| ICC/IF |
Human
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| WB |
Human
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| All applications |
Pig
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| Application | Abreviews | Notes |
|---|---|---|
| WB | (3) |
1/250. Detects a band of approximately 180 kDa (predicted molecular weight: 176 kDa).
Abcam recommends using milk as the blocking agent. |
| ICC/IF | (1) |
Use a concentration of 1 µg/ml.
|
| Notes |
|---|
|
WB
1/250. Detects a band of approximately 180 kDa (predicted molecular weight: 176 kDa). Abcam recommends using milk as the blocking agent. |
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ICC/IF
Use a concentration of 1 µg/ml. |
Target
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Function
Histone demethylase that specifically demethylates 'Lys-4' of histone H3, thereby playing a central role in histone code. Does not demethylate histone H3 'Lys-9', H3 'Lys-27', H3 'Lys-36', H3 'Lys-79' or H4 'Lys-20'. Demethylates trimethylated and dimethylated but not monomethylated H3 'Lys-4'. Participates in transcriptional repression of neuronal genes by recruiting histone deacetylases and REST at neuron-restrictive silencer elements. -
Tissue specificity
Expressed in all tissues examined. Highest levels found in brain and skeletal muscle. -
Involvement in disease
Defects in KDM5C are the cause of mental retardation syndromic X-linked JARID1C-related (MRXSJ) [MIM:300534]. MRXSJ is characterized by significantly sub-average general intellectual functioning associated with impairments in adaptative behavior and manifested during the developmental period. MRXSJ patients manifest mental retardation associated with variable features such as slowly progressive spastic paraplegia, seizures, facial dysmorphism. -
Sequence similarities
Belongs to the JARID1 histone demethylase family.
Contains 1 ARID domain.
Contains 1 JmjC domain.
Contains 1 JmjN domain.
Contains 2 PHD-type zinc fingers. -
Domain
The first PHD-type zinc finger domain recognizes and binds H3-K9Me3.
Both the JmjC domain and the JmjN domain are required for enzymatic activity. -
Cellular localization
Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 8242 Human
- Entrez Gene: 100037295 Pig
- Omim: 314690 Human
- SwissProt: P41229 Human
- Unigene: 631768 Human
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Alternative names
- DXS1272E antibody
- Histone demethylase JARID1C antibody
- JARID1C antibody
see all
Images
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Western blot - Anti-KDM5C / Jarid1C / SMCX antibody (ab34718)
Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: KDM5C knockout HAP1 whole cell lysate (20 µg)
Lane 3: HEK293 whole cell lysate (20 µg)
Lane 4: U2OS whole cell lysate (20 µg)Lanes 1 - 4: Merged signal (red and green). Green - ab34718 observed at 175 kDa. Red - loading control, ab18058, observed at 120 kDa.
ab34718 was shown to specifically recognize KDM5C in wild-type HAP1 cells as signal was lost at the expected MW in KDM5C knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and KDM5C knockout samples were subjected to SDS-PAGE. Ab34718 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1/250 dilution and 1/20,000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1/20,000 dilution for 1 hour at room temperature before imaging.
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Immunocytochemistry/ Immunofluorescence - Anti-KDM5C / Jarid1C / SMCX antibody (ab34718)PFA-fixed, 0.5% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells stained for KDM5C / Jarid1C / SMCX (green) using ab34718 at 1/200 dilution in ICC/IF. Counter-stained with DAPI in order to highlight the nucleus (red). Please refer to abreview for further experimental details.
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Immunocytochemistry/ Immunofluorescence - Anti-KDM5C / Jarid1C / SMCX antibody (ab34718)ab34718 stained in Hela cells. Cells were fixed with 100% methanol (5min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% triton for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab34718 at 5µg/ml and ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control) at 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150120 (pseudo-colored red) and ab150081 (colored green) used at 1 µg/ml for 1 hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43 µM for 1hour at room temperature.
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Western blot - Anti-KDM5C / Jarid1C / SMCX antibody (ab34718)All lanes : Anti-KDM5C / Jarid1C / SMCX antibody (ab34718) at 1/250 dilution
Lane 1 : HEK-293T cell lysate
Lane 2 : KDM5C knockout HEK-293T cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 176 kDa
Observed band size: 175 kDa why is the actual band size different from the predicted?Lanes 1 - 2: Merged signal (red and green). Green - ab34718 observed at 175 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.
ab34718 was shown to react with KDM5C / Jarid1C / SMCX in wild-type HEK-293T cells in western blot with loss of signal observed in KDM5C knockout cell line ab266251 (KDM5C knockout cell lysate ab257494). Wild-type and KDM5C knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab34718 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4°C at a 1 in 250 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Western blot - Anti-KDM5C / Jarid1C / SMCX antibody (ab34718)All lanes : Anti-KDM5C / Jarid1C / SMCX antibody (ab34718) at 1 µg/ml
Lane 1 : HEK293 (Human) Whole Cell Lysate
Lane 2 : Y79 (Human retinoblastoma cell line) Whole Cell Lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed at 1/50000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 176 kDa
Observed band size: 180 kDa why is the actual band size different from the predicted?
Additional bands at: 45 kDa, 58 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 20 minutesThis 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 3% Milk before being incubated with ab34718 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.
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Western blot - Anti-KDM5C / Jarid1C / SMCX antibody (ab34718)Anti-KDM5C / Jarid1C / SMCX antibody (ab34718) at 1/250 dilution +HEK-293 whole cell lysate (ab7902) at 20 µg
Secondary
IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 176 kDa
Observed band size: 176 kDa
Protocols
Datasheets and documents
References (6)
ab34718 has been referenced in 6 publications.
- Hong Z et al. KDM5C is transcriptionally regulated by BRD4 and promotes castration-resistance prostate cancer cell proliferation by repressing PTEN. Biomed Pharmacother 114:108793 (2019). PubMed: 30921702
- Wang J et al. Asymmetric Expression of LincGET Biases Cell Fate in Two-Cell Mouse Embryos. Cell 175:1887-1901.e18 (2018). PubMed: 30550787
- Fujita T et al. Identification of telomere-associated molecules by engineered DNA-binding molecule-mediated chromatin immunoprecipitation (enChIP). Sci Rep 3:3171 (2013). ICC/IF ; Human . PubMed: 24201379
- Liang Z et al. Proliferating cell nuclear antigen is required for loading of the SMCX/KMD5C histone demethylase onto chromatin. Epigenetics Chromatin 4:18 (2011). PubMed: 21996408
- Huang PH et al. Histone deacetylase inhibitors stimulate histone H3 lysine 4 methylation in part via transcriptional repression of histone H3 lysine 4 demethylases. Mol Pharmacol 79:197-206 (2011). WB, ChIP ; Human . PubMed: 20959362
- Kim JH & Youn HD C-terminal binding protein maintains mitochondrial activities. Cell Death Differ 16:584-92 (2009). ChIP ; Human . PubMed: 19136938
