WB
Western blot analysis of KDM2B on different lysates with Rabbit anti-KDM2B antibody at 1/2,000 dilution. Lane 1: HeLa cell lysate (20 µg/Lane) Lane 2: HepG2 cell lysate (20 µg/Lane) Lane 3: 293T cell lysate (20 µg/Lane) Lane 4: NIH/3T3 cell lysate (20 µg/Lane) Lane 5: C2C12 cell lysate (20 µg/Lane) Lane 6: PC-12 cell lysate (20 µg/Lane) Lane 7: C6 cell lysate (20 µg/Lane) Lane 8: Human brain tissue lysate (40 µg/Lane) Lane 9: Mouse brain tissue lysate (40 µg/Lane) Lane 10: Mouse colon tissue lysate (40 µg/Lane) Lane 11: Rat brain tissue lysate (40 µg/Lane) Exposure time: 25 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-KDM2B antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.ICC/IF
Immunocytochemistry analysis of HeLa cells labeling KDM2B with Rabbit anti-KDM2B antibody at 1/2,000 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-KDM2B antibody at 1/2,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.FC
Flow cytometric analysis of NIH/3T3 cells labeling KDM2B. Cells were fixed and permeabilized. Then stained with the primary antibody (1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).IP
KDM2B was immunoprecipitated from 0.2 mg MDA-MB-231 cell lysate withprimary antibody at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using primary antibody at 1/2,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: MDA-MB-231 cell lysate (input) Lane 2:primary antibody IP in MDA-MB-231 cell lysate Lane 3: Rabbit IgG instead of primary antibody in MDA-MB-231 cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 10 seconds.ChIP
Chromatin immunoprecipitations were performed with cross-linked chromatin from MDA-MB-231 cells with KDM2B or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.| Product Name | KDM2B Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Recombinant protein within mouse KDM2B aa 681-920 / 1,309. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | ChIPFCICC/IFIHCIPWB |
| WB:1:2000 IHC:1:200-1:1000 ICC:1:2000 FC:1:1000 IP:1-2μg/sample ChIP:Use 5 μg for 25 μg of chromatin. |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | KDM2B |
|---|---|
| Gene Synonyms | CXXC2 Fbl10 PCCX2 FBXL10 JHDM1B |
| Gene Full Name | lysine demethylase 2B |
| Gene Summary | This gene encodes a member of the F-box protein family which is characterized by an approximately 40 amino acid motif, the F-box. The F-box proteins constitute one of the four subunits of ubiquitin protein ligase complex called SCFs (SKP1-cullin-F-box), which function in phosphorylation-dependent ubiquitination. The F-box proteins are divided into 3 classes: Fbws containing WD-40 domains, Fbls containing leucine-rich repeats, and Fbxs containing either different protein-protein interaction modules or no recognizable motifs. The protein encoded by this gene belongs to the Fbls class. Multiple alternatively spliced transcript variants have been found for this gene, but the full-length nature of some variants has not been determined. [provided by RefSeq, Jul 2008] |
| 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 [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 |
| Molecular Weight(MW) | 150kDa |
| Cellular Localization | Nucleus, nucleolus, Chromosome. |
WB
Western blot analysis of KDM2B on different lysates with Rabbit anti-KDM2B antibody at 1/2,000 dilution. Lane 1: HeLa cell lysate (20 µg/Lane) Lane 2: HepG2 cell lysate (20 µg/Lane) Lane 3: 293T cell lysate (20 µg/Lane) Lane 4: NIH/3T3 cell lysate (20 µg/Lane) Lane 5: C2C12 cell lysate (20 µg/Lane) Lane 6: PC-12 cell lysate (20 µg/Lane) Lane 7: C6 cell lysate (20 µg/Lane) Lane 8: Human brain tissue lysate (40 µg/Lane) Lane 9: Mouse brain tissue lysate (40 µg/Lane) Lane 10: Mouse colon tissue lysate (40 µg/Lane) Lane 11: Rat brain tissue lysate (40 µg/Lane) Exposure time: 25 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-KDM2B antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ICC/IF
Immunocytochemistry analysis of HeLa cells labeling KDM2B with Rabbit anti-KDM2B antibody at 1/2,000 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-KDM2B antibody at 1/2,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.
FC
Flow cytometric analysis of NIH/3T3 cells labeling KDM2B. Cells were fixed and permeabilized. Then stained with the primary antibody (1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
IP
KDM2B was immunoprecipitated from 0.2 mg MDA-MB-231 cell lysate withprimary antibody at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using primary antibody at 1/2,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: MDA-MB-231 cell lysate (input) Lane 2:primary antibody IP in MDA-MB-231 cell lysate Lane 3: Rabbit IgG instead of primary antibody in MDA-MB-231 cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 10 seconds.
ChIP
Chromatin immunoprecipitations were performed with cross-linked chromatin from MDA-MB-231 cells with KDM2B or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.| Application Notes | WB:1:2000 IHC:1:200-1:1000 ICC:1:2000 FC:1:1000 IP:1-2μg/sample ChIP:Use 5 μg for 25 μg of chromatin. |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
