Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human, Mouse

Cited:

Human, Mouse

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Immunocytochemistry/ Immunofluorescence, Simple Western, Immunoprecipitation, Chromatin Immunoprecipitation, Chromatin Immunoprecipitation (ChIP), Knockdown Validated

Cited:

Immunohistochemistry-Paraffin, Western Blot, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, Chemotaxis

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all Lysine (K)-specific Demethylase 4C/KDM4C Primary Antibodies »

Product Specifications

Immunogen

A partial recombinant protein made to an internal region of the human JMJD2C protein (within residues 450-600). [Swiss-Prot Q9H3R0]

Localization

Nucleus

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for Lysine (K)-specific Demethylase 4C/KDM4C/JMJD2C Antibody - BSA Free

Western Blot: Lysine (K)-specific Demethylase 4C/KDM4C/JMJD2C AntibodyBSA Free [NBP1-49600]

Western Blot: Lysine (K)-specific Demethylase 4C/KDM4C/JMJD2C AntibodyBSA Free [NBP1-49600]

Western Blot: Lysine (K)-specific Demethylase 4C/KDM4C/JMJD2C Antibody [NBP1-49600] - Analysis of JMJD2C in HeLa nuclear extracts.
Immunocytochemistry/ Immunofluorescence: Lysine (K)-specific Demethylase 4C/KDM4C/JMJD2C Antibody - BSA Free [NBP1-49600]

Immunocytochemistry/ Immunofluorescence: Lysine (K)-specific Demethylase 4C/KDM4C/JMJD2C Antibody - BSA Free [NBP1-49600]

Immunocytochemistry/Immunofluorescence: Lysine (K)-specific Demethylase 4C/KDM4C/JMJD2C Antibody [NBP1-49600] - NIH3T3 cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.5% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti-Lysine (K)-specific Demethylase 4C/KDM4C/JMJD2C Antibody NBP1-49600 at 2 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.
Immunohistochemistry-Paraffin: Lysine (K)-specific Demethylase 4C/KDM4C/JMJD2C Antibody - BSA Free [NBP1-49600]

Immunohistochemistry-Paraffin: Lysine (K)-specific Demethylase 4C/KDM4C/JMJD2C Antibody - BSA Free [NBP1-49600]

Immunohistochemistry-Paraffin: Lysine (K)-specific Demethylase 4C/KDM4C/JMJD2C Antibody [NBP1-49600] - Staining of JMJD2C in paraffin embedded mouse pancreas using DAB with hematoxylin counterstain.
Immunocytochemistry/ Immunofluorescence: Lysine (K)-specific Demethylase 4C/KDM4C/JMJD2C Antibody - BSA Free [NBP1-49600]

Immunocytochemistry/ Immunofluorescence: Lysine (K)-specific Demethylase 4C/KDM4C/JMJD2C Antibody - BSA Free [NBP1-49600]

Immunocytochemistry/Immunofluorescence: Lysine (K)-specific Demethylase 4C/KDM4C/JMJD2C Antibody [NBP1-49600] - HeLa cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.5% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti-Lysine (K)-specific Demethylase 4C/KDM4C/JMJD2C Antibody NBP1-49600 at 2 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.
Simple Western: Lysine (K)-specific Demethylase 4C/KDM4C/JMJD2C AntibodyBSA Free [NBP1-49600]

Simple Western: Lysine (K)-specific Demethylase 4C/KDM4C/JMJD2C AntibodyBSA Free [NBP1-49600]

Simple Western: Lysine (K)-specific Demethylase 4C/KDM4C/JMJD2C Antibody [NBP1-49600] - Simple Western lane view shows a specific band for JMJD2C in 1.0 mg/ml of HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.
Lysine (K)-specific Demethylase 4C/KDM4C/JMJD2C Antibody - BSA Free

Western Blot: Rabbit Polyclonal Lysine (K)-specific Demethylase 4C/KDM4C/JMJD2C Antibody [NBP1-49600]

Western Blot: Rabbit Polyclonal Lysine (K)-specific Demethylase 4C/KDM4C/JMJD2C Antibody [NBP1-49600] - shRNA Knockdown using 4C/KDM4C/JMJD2C Antibody. Image from a verified customer review.
Lysine (K)-specific Demethylase 4C/KDM4C/JMJD2C Antibody - BSA Free

Western Blot: Lysine (K)-specific Demethylase 4C/KDM4C/JMJD2C Antibody - BSA Free [NBP1-49600] -

KDM4C, but not KDM4B, depletion increases chromosomal segregation errors during mitosis. (A)–(D) Representative images showing normal and defective mitotic U2OS cells depleted of KDM4C. Cells were subjected to immunofluorescence analysis using Pericentrin (green) and alpha -tubulin antibodies (red). DNA is stained with DAPI (blue). (A) Normal and abnormal metaphase with misaligned chromosome (indicated by white arrow). (B) Normal and abnormal anaphase with either lagging chromosomes or anaphase bridge (indicated by white arrows). (C) Abnormal telophase with either lagging chromosomes or telophase bridge (indicated by white arrows). (D) Multipolar metaphase. (E) KDM4B and KDM4C knockdown by western blotting. U2OS cells were transfected with either control or different sequences of KDM4B and KDM4C Stealth siRNA (Invitrogen). Protein extracts were prepared 72 h after transfection and immunoblotted with KDM4B and KDM4C antibody. beta -Actin is used as a loading control. (F) A histogram showing the percentage of metaphases with misaligned chromosomes 72 h after transfection with control and different KDM4B-C siRNA sequences. KDM4C, but not KDM4B, depletion increases the frequency of metaphase cells with misaligned chromosomes. n, number of metaphase cells counted. Error bars represent standard deviation from two independent experiments. (G) KDM4C, but not KDM4B, depletion increases the frequency of anaphase–telophase cells with either lagging chromosomes or anaphase–telophase bridges. As in (F), except that the histogram shows the percentage of defective anaphase–telophase cells. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/24728997), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for Lysine (K)-specific Demethylase 4C/KDM4C/JMJD2C Antibody - BSA Free

Application
Recommended Usage

Chromatin Immunoprecipitation

reported in scientific literature (PMID 23884959; 23129632)

Immunocytochemistry/ Immunofluorescence

1:100

Immunohistochemistry

1:100

Immunohistochemistry-Paraffin

1:100

Immunoprecipitation

reported in scientific literature (PMID 23129632)

Knockdown Validated

Validated for Knockdown from a verified customer review.

Simple Western

1:12.5

Western Blot

1:1000
Application Notes
In Western blot, a band is seen ~119 kDa in HeLa cells. In ICC/IF, nuclear staining was observed in HeLa cells. In IHC-P, staining was observed in the nuclei of mouse pancreas. Prior to immunostaining paraffin tissues, antigen retrieval with sodium citrate buffer (pH 6.0) is recommended.

In Simple Western only 10 - 15 uL of the recommended dilution is used per data point.
See Simple Western Antibody Database for Simple Western validation: Tested in HeLa lysate 1.0 mg/mL, separated by Size, antibody dilution of 1:12.5, apparent MW was 158 kDa. Separated by Size-Wes, Sally Sue/Peggy Sue.

Reviewed Applications

Read 2 reviews rated 4.5 using NBP1-49600 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS

Format

BSA Free

Preservative

0.05% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: Lysine (K)-specific Demethylase 4C/KDM4C

JMJD2C (Jumonji C domain-containing oxygenase D2C; also called GASC1, JHDM3C or KDM4C) belongs to JMJD2 family and is a histone demethylase for di- and trimethylated lysine 9 and 36 on histone H3 (H3K9me3/2 and H3K36me3/2). H3K9me3/2 is generally associated with transcriptional repression and heterochromatin formation, whereas, H3K36me3/2 is linked to transcriptionaly active genes where it plays a key role in suppression of incorrect transcription. JMJD2C regulates the expression of pluripotency genes including NOTCH1, NANOG, Sox2 and Pou5, and because of its dual role in modifying H3 either by removing the repressive H3K9me3/2 or the active H3K36me3/2 factor, JMJD2C has been considered a fine-tuning regulator of gene expression in normal development/differentiation as well as in carcinogenic progression. JMJD2C is overexpressed in aggressive, basal-like breast cancers compared with non basal-like breast cancers, and its overexpression has been shown to cause transformation of immortalized, non-transformed mammary epithelial cells and to regulate expression of genes responsible for stem cell phenotype/self-renewal in breast cancer cells. Moreover oncogenic role of JMJD2C has been documented in prostate cancer where it enhances the transcription of AR-dependent genes and cell proliferation by interaction with ligand-bound AR.

Alternate Names

GASC1, JHDM3C, JMJD2C, KDM4C, Lysine (K)specific Demethylase 4C

Entrez Gene IDs

23081 (Human); 76804 (Mouse)

Gene Symbol

KDM4C

UniProt

Q9H3R0 (Human), Q8VCD7 (Mouse)

Additional Lysine (K)-specific Demethylase 4C/KDM4C Products

Product Documents for Lysine (K)-specific Demethylase 4C/KDM4C/JMJD2C Antibody - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Download SDS

Product Specific Notices for Lysine (K)-specific Demethylase 4C/KDM4C/JMJD2C Antibody - BSA Free

Manufactured by Genomic Antibody Technology™. GAT FAQs

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Related Research Areas

Epigenetics

Citations for Lysine (K)-specific Demethylase 4C/KDM4C/JMJD2C Antibody - BSA Free

Customer Reviews for Lysine (K)-specific Demethylase 4C/KDM4C/JMJD2C Antibody - BSA Free (2)

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  • Lysine (K)-specific Demethylase 4C/KDM4C/JMJD2C Antibody - BSA Free
    Name: Anonymous
    Application: Western Blot
    Sample Tested: HEK293T cells
    Species: Human
    Verified Customer | Posted 10/15/2024
    KDM4C antibody and shRNA knockdown efficiency
    Lysine (K)-specific Demethylase 4C/KDM4C/JMJD2C Antibody - BSA Free NBP1-49600
  • Name: Anonymous
    Application: Western Blot
    Sample Tested: HeLa lysates
    Species: Human
    Verified Customer | Posted 05/03/2012
    Lysine (K)-specific Demethylase 4C/KDM4C/JMJD2C Antibody - BSA Free NBP1-49600

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Protocols

View specific protocols for Lysine (K)-specific Demethylase 4C/KDM4C/JMJD2C Antibody - BSA Free (NBP1-49600):


Immunocytochemistry Protocol

Culture cells to appropriate density in 35 mm culture dishes or 6-well plates.

1. Remove culture medium and add 10% formalin to the dish. Fix at room temperature for 30 minutes.
2. Remove the formalin and add ice cold methanol. Incubate for 5-10 minutes.
3. Remove methanol and add washing solution (i.e. PBS). Be sure to not let the specimen dry out. Wash three times for 10 minutes.
4. To block nonspecific antibody binding incubate in 10% normal goat serum from 1 hour to overnight at room temperature.
5. Add primary antibody at appropriate dilution and incubate at room temperature from 2 hours to overnight at room temperature.
6. Remove primary antibody and replace with washing solution. Wash three times for 10 minutes.
7. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
8. Remove antibody and replace with wash solution, then wash for 10 minutes. Add Hoechst 33258 to wash solution at 1:25,0000 and incubate for 10 minutes. Wash a third time for 10 minutes.
9. Cells can be viewed directly after washing. The plates can also be stored in PBS containing Azide covered in Parafilm (TM). Cells can also be cover-slipped using Fluoromount, with appropriate sealing.

*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.


Immunohistochemistry-Paraffin Embedded Sections

Antigen Unmasking:
Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes.

Staining:
1. Wash sections in deionized water three times for 5 minutes each.
2. Wash sections in wash buffer for 5 minutes.
3. Block each section with 100-400 ul blocking solution for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul diluted primary antibody. Incubate overnight at 4C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul biotinylated diluted secondary antibody. Incubate 30 minutes at room temperature.
7. Remove secondary antibody solution and wash sections three times with wash buffer for 5 minutes each.
8. Add 100-400 ul Streptavidin-HRP reagent to each section and incubate for 30 minutes at room temperature.
9. Wash sections three times in wash buffer for 5 minutes each.
10. Add 100-400 ul DAB substrate to each section and monitor staining closely.
11. As soon as the sections develop, immerse slides in deionized water.
12. Counterstain sections in hematoxylin.
13. Wash sections in deionized water two times for 5 minutes each.
14. Dehydrate sections.
15. Mount coverslips.


Western Blot Protocol

1. Perform SDS-PAGE on samples to be analyzed, loading 40 ug of total protein per lane.
2. Transfer proteins to membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
3. Stain according to standard Ponceau S procedure (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
4. Rinse the blot.
5. Block the membrane using standard blocking buffer for at least 1 hour.
6. Wash the membrane in wash buffer three times for 10 minutes each.
7. Dilute the rabbit anti-JMJD2C primary antibody (NBP1-49600) in blocking buffer and incubate 1 hour at room temperature.
8. Wash the membrane in wash buffer three times for 10 minutes each.
9. Apply diluted rabbit-IgG HRP-conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature.
10. Wash the blot in wash buffer three times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturers instructions.

*Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2%.

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for Lysine (K)-specific Demethylase 4C/KDM4C/JMJD2C Antibody - BSA Free

Showing  1 - 2 of 2 FAQs Showing All

  • Q: How has this antibody been tested for reactivity with mouse other than immunohistochemistry?

    A:

    We do not have data for NBP1-49600 with mouse other than in IHC-P. However, if you use NBP1-49600 to analyze your mouse samples in any of the validated applications (those stated on the datasheet) for this antibody, you would be covered by our Novus Guarantee, which means that if you did have any issues, we would resolve them.

  • Q: I am interested in the KDM4cJMJD2c antibody NBP1-49600. According to their website, it is used against human and murine JMJD2c protein. In addition, there is an image of an Immunhisto (Mouse pancreas).We need the antibody for the main part for Western blots with mouse protein. Are there positive reviews about it? Did they specify the reactivity? How specific does it recognize the protein? Does it recognize the protein properly? For an antibody of a different company, we already had the problem that mouse was specified on their datasheet but in the end wasn't recognized.

    A: This antibody, being a polyclonal does recognize both the human and mouse proteins. It has been shown to work much better on the native mouse proteins in IHC than in Western blot. It does recognize mouse JMJD2C in Western blot but we have seen that it does give some background staining. Regardless, we have a 100% guarantee on all of our products for the applications and species listed on our datasheet, so we will guarantee this for Western blot on mouse samples.

  • Q: How has this antibody been tested for reactivity with mouse other than immunohistochemistry?

    A:

    We do not have data for NBP1-49600 with mouse other than in IHC-P. However, if you use NBP1-49600 to analyze your mouse samples in any of the validated applications (those stated on the datasheet) for this antibody, you would be covered by our Novus Guarantee, which means that if you did have any issues, we would resolve them.

  • Q: I am interested in the KDM4cJMJD2c antibody NBP1-49600. According to their website, it is used against human and murine JMJD2c protein. In addition, there is an image of an Immunhisto (Mouse pancreas).We need the antibody for the main part for Western blots with mouse protein. Are there positive reviews about it? Did they specify the reactivity? How specific does it recognize the protein? Does it recognize the protein properly? For an antibody of a different company, we already had the problem that mouse was specified on their datasheet but in the end wasn't recognized.

    A: This antibody, being a polyclonal does recognize both the human and mouse proteins. It has been shown to work much better on the native mouse proteins in IHC than in Western blot. It does recognize mouse JMJD2C in Western blot but we have seen that it does give some background staining. Regardless, we have a 100% guarantee on all of our products for the applications and species listed on our datasheet, so we will guarantee this for Western blot on mouse samples.

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