KDM2A/FBXL11 Antibody - BSA Free

Novus Biologicals | Catalog # NBP1-78305

Novus Biologicals
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Key Product Details

Species Reactivity

Human, Mouse

Applications

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

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all KDM2A/FBXL11 Primary Antibodies »

Product Specifications

Immunogen

A synthetic peptide made to an N-terminal portion of the human KDM2A/FBXL11 protein (between residues 350-500) [UniProt Q9Y2K7]

Localization

Nucleus, nucleoplasm. Note: Punctate expression throughout the nucleoplasm and enriched in the perinucleolar region. Specifically nucleates at CpG islands where it's presence results in chromatin depleted in H3K36me2.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for KDM2A/FBXL11 Antibody - BSA Free

Western Blot: KDM2A/FBXL11 AntibodyBSA Free [NBP1-78305]

Western Blot: KDM2A/FBXL11 AntibodyBSA Free [NBP1-78305]

Western Blot: KDM2A/FBXL11 Antibody [NBP1-78305] - 1) 293T. 2) 293T transiently transfected to express human KDM2A. WB image submitted by a verified customer review.
Immunocytochemistry/ Immunofluorescence: KDM2A/FBXL11 Antibody - BSA Free [NBP1-78305]

Immunocytochemistry/ Immunofluorescence: KDM2A/FBXL11 Antibody - BSA Free [NBP1-78305]

Immunocytochemistry/Immunofluorescence: KDM2A/FBXL11 Antibody - BSA Free [NBP1-78305] - Mouse MS1 cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with KCM2A/FBXL11 Antibody (NBP1-78305) at 2ug/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: KDM2A/FBXL11 Antibody - BSA Free [NBP1-78305]

Immunohistochemistry: KDM2A/FBXL11 Antibody - BSA Free [NBP1-78305]

Immunohistochemistry: KDM2A/FBXL11 Antibody [NBP1-78305] - IHC analysis of KDM2A / FBXL11 in human kidney cancer xenograft using DAB with hematoxylin counterstain.
Immunocytochemistry/ Immunofluorescence: KDM2A/FBXL11 Antibody - BSA Free [NBP1-78305]

Immunocytochemistry/ Immunofluorescence: KDM2A/FBXL11 Antibody - BSA Free [NBP1-78305]

Immunocytochemistry/Immunofluorescence: KDM2A/FBXL11 Antibody [NBP1-78305] - Human astrocytoma cell line lentivirally transduced and expressing KDM2A. Cells were fixed with 4% paraformaldehyde and stained with anti-KDM2A, followed by Alexa Fluor 488-anti-rabbit secondary antibody. DAPI staining shows nuclei. ICC/IF image submitted by a verified customer review.
Immunocytochemistry/ Immunofluorescence: KDM2A/FBXL11 Antibody - BSA Free [NBP1-78305]

Immunocytochemistry/ Immunofluorescence: KDM2A/FBXL11 Antibody - BSA Free [NBP1-78305]

Immunocytochemistry/Immunofluorescence: KDM2A/FBXL11 Antibody - BSA Free [NBP1-78305] - Caco-2 cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with KDM2A/FBXL11 Antibody (NBP1-78305) at 1ug/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 40X objective.

Applications for KDM2A/FBXL11 Antibody - BSA Free

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:100-1:400

Immunohistochemistry

1:300

Immunohistochemistry-Paraffin

1:300

Western Blot

reported by customer review
Application Notes
This KDM2A/FBXL11 antibody is useful for Immunocytochemistry/Immunofluorescence where nuclear staining is observed in HeLa cells and IHC-paraffin embedded sections where nuclear staining is seen in human kidney cancer xenograft. Prior to immunostaining paraffin tissues, antigen retrieval with sodium citrate buffer (pH 6.0) is recommended.

Reviewed Applications

Read 3 reviews rated 4.7 using NBP1-78305 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS and 30% Glycerol

Format

BSA Free

Preservative

0.05% Sodium Azide

Concentration

1.2 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: KDM2A/FBXL11

KDM2A / FBXL11 (also known as JHDM1A, lysine specific demethylase 1A) is a F-box family member and JmjC-domain containing histone demethylase that specifically demethylates Lysine 36 of H3 (H3 K36). KDM2A / FBXL11 is involved in epigenetic silencing and required to maintain the heterochromatic state, as well as sustaining centromeric integrity and genomic stability during mitosis (PMID 19001877). Under conditions of starvation, FBXL11's ability to repress RNA transcription has been shown to be reduced (PMID 20379134). In addition, KDM2A / FBXL11 is also a key inhibitory regulator of NF kappa B activity. This regulation occurs when K218 and K221 of the p65 subunit are methylated. The overexpression of KDM2A / FBXL11 can decrease the viability of specific cancer cell lines, while downregulation was shown to have the opposite effect (PMID 20080798). KDM2A / FBXL11 shows further regulatory involvement at CpG islands. Nonmethylated CpG islands preferentially bind KDM2A / FBXL11 using a zinc finger domain, in which the binding is blocked when CpG DNA is in its methlylated form (PMID 20417597).

Long Name

Lysine-specific demethylase 2A

Alternate Names

CXXC8, EC 1.14.11.27, F-box protein FBL7, FBL11, FBL7, FBXL11, JHDM1A, KDM2A, KIAA1004

Entrez Gene IDs

22992 (Human)

Gene Symbol

KDM2A

UniProt

Q9Y2K7 (Human)

Additional KDM2A/FBXL11 Products

Product Documents for KDM2A/FBXL11 Antibody - BSA Free

Certificate of Analysis

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Product Specific Notices for KDM2A/FBXL11 Antibody - BSA Free

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.

Customer Reviews for KDM2A/FBXL11 Antibody - BSA Free (3)

4.7 out of 5
3 Customer Ratings
5 Stars
67%
4 Stars
33%
3 Stars
0%
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1 Stars
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Customer Images

Showing  1 - 3 of 3 reviews Showing All
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  • KDM2A/FBXL11 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: 293T
    Species: Human
    Verified Customer | Posted 12/31/2019
    1. 293T 2. 293T transiently transfected to express human KDM2A
    KDM2A/FBXL11 Antibody - BSA Free NBP1-78305
  • KDM2A/FBXL11 Antibody
    Name: Anonymous
    Application: Immunocytochemistry
    Sample Tested: AGS human gastric adenocarcinoma cell line and brain tumor cell line
    Species: Human
    Verified Customer | Posted 12/31/2019
    Human astrocytoma cell line lentivirally transduced and expressing KDM2A. Cells were fixed with 4% paraformaldehyde and stained with anti-KDM2A, followed by Alexa488-anti-rabbit secondary antibody. DAPI staining shows nuclei.
    KDM2A/FBXL11 Antibody - BSA Free NBP1-78305
  • Name: Anonymous
    Application: Western Blot
    Sample Tested: Human lung cancer cell lines whole cell lysate
    Species: Human
    Verified Customer | Posted 05/18/2016
    Western blot shows detection of Rabbit Anti-Human KDM2A polyclonal Antibody in lung cancer cell lines (dilution 1:1000)
    KDM2A/FBXL11 Antibody - BSA Free NBP1-78305

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Protocols

View specific protocols for KDM2A/FBXL11 Antibody - BSA Free (NBP1-78305):

KDM2A/FBXL11 Antibody:
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.

KDM2A/FBXL11 Antibody:
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.
*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.

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FAQs

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