Overview

Properties

Applications

Our Abpromise guarantee covers the use of ab5199 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-P Use a concentration of 1 - 3 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Target

  • Function
    Histone demethylase that demethylates 'Lys-4' and 'Lys-36' of histone H3, thereby playing a central role in histone code. Preferentially demethylates trimethylated H3 'Lys-4' and dimethylated H3 'Lys-36' residue while it has weak or no activity for mono- and tri-methylated H3 'Lys-36'. Preferentially binds the transcribed region of ribosomal RNA and represses the transcription of ribosomal RNA genes which inhibits cell growth and proliferation. May also serve as a substrate-recognition component of the SCF (SKP1-CUL1-F-box protein)-type E3 ubiquitin ligase complex.
  • Sequence similarities
    Belongs to the JHDM1 histone demethylase family.
    Contains 1 CXXC-type zinc finger.
    Contains 1 F-box domain.
    Contains 1 JmjC domain.
    Contains 7 LRR (leucine-rich) repeats.
    Contains 1 PHD-type zinc finger.
  • Domain
    The JmjC domain mediates demethylation activity (By similarity). It is also required for repression of ribosomal RNA genes.
  • Cellular localization
    Nucleus > nucleolus.
  • Information by UniProt
  • Database links
  • Alternative names
    • [Histone-H3]-lysine-36 demethylase 1B antibody
    • CXXC-type zinc finger protein 2 antibody
    • CXXC2 antibody
    • F box and leucine rich repeat protein 10 antibody
    • F box protein FBL10 antibody
    • F-box and leucine-rich repeat protein 10 antibody
    • F-box protein FBL10 antibody
    • F-box/LRR-repeat protein 10 antibody
    • Fbl10 antibody
    • FBXL10 antibody
    • JEMMA (Jumonji domain EMSY interactor methyltransferase motif) protein antibody
    • JHDM1B antibody
    • JmjC domain-containing histone demethylation protein 1B antibody
    • Jumonji C domain containing histone demethylase 1B antibody
    • Jumonji domain-containing EMSY-interactor methyltransferase motif protein antibody
    • kdm2b antibody
    • KDM2B_HUMAN antibody
    • Lysine (K) specific demethylase 2B antibody
    • Lysine-specific demethylase 2B antibody
    • PCCX2 antibody
    • Protein containing CXXC domain 2 antibody
    • Protein JEMMA antibody
    • Protein-containing CXXC domain 2 antibody
    see all

References

This product has been referenced in:
  • Zhang DD  et al. Vildagliptin, a DPP4 inhibitor, alleviates diabetes-associated cognitive deficits by decreasing the levels of apoptosis-related proteins in the rat hippocampus. Exp Ther Med 15:5100-5106 (2018). Read more (PubMed: 29805536) »
  • Tzatsos A  et al. Lysine-specific Demethylase 2B (KDM2B)-let-7-Enhancer of Zester Homolog 2 (EZH2) Pathway Regulates Cell Cycle Progression and Senescence in Primary Cells. J Biol Chem 286:33061-9 (2011). WB ; Mouse . Read more (PubMed: 21757686) »
See all 3 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Question

LOT NUMBER 626682 ORDER NUMBER 2 (dated 05/01/2011) DESCRIPTION OF THE PROBLEM High background, more bands SAMPLE MCF-7 and ZR-75-1 human cell lines whole cell protein extracts PRIMARY ANTIBODY Abcam Goat anti JHDM1B diluted 1/1000 or 1/2000 in TBS-Tween + 3.5% BSA, or 2% blocking agent as above in TBS-T, or 2% Normal Goat serum in TBS-T, O/N @ 4°C, followed by 3 washes of 10' each in TBS-Tween at RT DETECTION METHOD ECL Advance from GE healthcare POSITIVE AND NEGATIVE CONTROLS USED None ANTIBODY STORAGE CONDITIONS -20°C as specified by spec sheet SAMPLE PREPARATION Proteins extracted in: Igepal 1% KH2PO4 0.1M pH7.5 0.1mM Glycerophosphate (standard buffer in our lab) added protease inhibitors mix added Laemli loading buffer and heated for 10' 95°C AMOUNT OF PROTEIN LOADED 15 micrograms ELECTROPHORESIS/GEL CONDITIONS SDS-PAGE 6%, run through stacking at 50V and through resolving at 150V Standard TGS Running buffer TRANSFER AND BLOCKING CONDITIONS Standard Tris/Glycine/SDS transfer buffer, 280mA 2Hours 4°C. Blocking agent: tried 2% normal goat serum in TBS-Tween 1h @ RT or 2% GE Health Care Advance ECL Kit Blocking agent in TBS-Tween (as recommended by manufacturer: 1h @ RT) SECONDARY ANTIBODY Santa Cruz donkey anti Goat dilutent: as above for primary antibody 1/2500, incubation 1h @ RT, followed by 3 washes of 10' each in TBS-Tween at RT HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 6 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? No WHAT STEPS HAVE YOU ALTERED? Blocking conditions and primary/secondary antibody dilution and incubation conditions

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Answer

Thank you for contacting Abcam Technical Team and for taking the time to provide some useful details of the experiments. I am very sorry to hear that you are having problems with this product. As our Abpromise indicates, in the event that a product is not functioning in the applications/species cited on the product data sheet (and the problem has been reported within 6 months of purchase) we will happily offer a credit note/refund to the value of the product purchased. After reading through the detailed protocol you kindly forwarded to Abcam, I would like to make the following comments/suggestions: 1) Isoform: It is important to consider that at least 3 isoforms (Q8NHM5, Q8NHM5-2, Q8NHM5-3) have been described which may run differently on the gel. 2) Sample preparation: KDM2B is expressed mainly in the nucleoli, so it may be worth preparing nuclear lysate (rather than whole cell lysate) for Western blot application. 3) Positive control: This antibody has been tested on cell cell lyaste of HeLa, so running this type of lysate alongside with the samples - as positive control - may be useful. 4) Detections system: Does the detection system work fine? Have you used it successfully with another primary antibody? Have you run a no primary - only secondary antibody - control to see if any of the non-specific bands are due to the secondary or not? If you have not done yet, I would advise you to check it. I hope this will be useful for you.

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Abreviews
Application
Western blot
Sample
Mouse Cell lysate - other (NIH3T3, Neuro2A and ES)
Loading amount
20 µg
Specification
NIH3T3, Neuro2A and ES
Gel Running Conditions
Reduced Denaturing (8)
Blocking step
BSA as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Jun 28 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa cells)
Loading amount
100 µg
Specification
HeLa cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10%

Abcam user community

Verified customer

Submitted Jul 26 2006

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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