Rabbit Recombinant Monoclonal CBX4 antibody. Suitable for IP, ChIP, WB, Flow Cyt (Intra), IHC-P and reacts with Human, Mouse, Rat samples. Cited in 3 publications.
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
IP | ChIP | WB | ICC/IF | Flow Cyt (Intra) | IHC-P | IHC-Fr | |
---|---|---|---|---|---|---|---|
Human | Tested | Tested | Tested | Not recommended | Tested | Not recommended | Not recommended |
Mouse | Expected | Expected | Tested | Not recommended | Expected | Tested | Not recommended |
Rat | Expected | Expected | Tested | Not recommended | Expected | Tested | Not recommended |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/30 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 2 µg for 25.00000 µg chromatin | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/1000 | Notes - |
Species Rat | Dilution info 1/1000 | Notes - |
Species Human | Dilution info 1/1000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human, Mouse, Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/600 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/200 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Rat | Dilution info 1/200 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info - | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Rat, Mouse, Human | Dilution info - | Notes - |
Select an associated product type
E3 SUMO-protein ligase which facilitates SUMO1 conjugation by UBE2I (PubMed:12679040). Involved in the sumoylation of HNRNPK, a p53/TP53 transcriptional coactivator, hence indirectly regulates p53/TP53 transcriptional activation resulting in p21/CDKN1A expression. Monosumoylates ZNF131 (PubMed:22825850).Component of a Polycomb group (PcG) multiprotein PRC1-like complex, a complex class required to maintain the transcriptionally repressive state of many genes, including Hox genes, throughout development (PubMed:12167701, PubMed:19636380, PubMed:21282530). PcG PRC1 complex acts via chromatin remodeling and modification of histones; it mediates monoubiquitination of histone H2A 'Lys-119', rendering chromatin heritably changed in its expressibility (PubMed:12167701, PubMed:19636380, PubMed:21282530). Binds to histone H3 trimethylated at 'Lys-9' (H3K9me3) (By similarity). Plays a role in the lineage differentiation of the germ layers in embryonic development (By similarity).
E3 SUMO-protein ligase CBX4, Chromobox protein homolog 4, Polycomb 2 homolog, Pc2, hPc2, CBX4
Rabbit Recombinant Monoclonal CBX4 antibody. Suitable for IP, ChIP, WB, Flow Cyt (Intra), IHC-P and reacts with Human, Mouse, Rat samples. Cited in 3 publications.
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
EPR23053-7
Affinity purification Protein A
The human recommendation is based on the WB results. We do not guarantee IHC-P for human.
Blue Ice
1-2 weeks
+4°C
-20°C
Upon delivery aliquot
Avoid freeze / thaw cycle
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
This supplementary information is collated from multiple sources and compiled automatically.
CBX4 also known as Chromobox protein homolog 4 or Polycomb 2 acts mechanically as part of the Polycomb group (PcG) proteins. It weighs approximately 60 kDa and it is found in nuclear matrix where it participates in chromatin remodeling. CBX4 shows significant expression in tissues like liver brain and lung indicating its wide importance to cellular function. By binding to methylated histones especially H3K27me3 CBX4 influences gene silencing and epigenetic regulation important for maintaining cellular identity.
CBX4 operates as a component of the Polycomb Repressive Complex 1 (PRC1) which plays role in controlling gene expression patterns during development. In this complex CBX4 interacts with other core components such as BMI1 and RING1 to facilitate monoubiquitination of histone H2A leading to transcriptional repression. This ability of CBX4 to modulate epigenetic states highlights its importance in critical developmental processes and differentiation.
CBX4 significantly influences gene regulatory networks involved in cell cycle and apoptosis pathways. In the cell cycle regulation CBX4 interacts with important proteins such as Cyclin D1 impacting cell proliferation. In apoptosis CBX4 has roles linked to the p53 pathway by modulating transcriptional activity which influences cellular fate. These pathways underline the significance of CBX4 in maintaining balance between cell growth and death.
The deregulation of CBX4 expression has been associated with cancers like hepatocellular carcinoma and breast cancer. CBX4 potentially interacts with proteins such as p21 and p16 whose pathways play significant roles in cancer development. This underlines its role as a potential biomarker or therapeutic target in cancer treatment strategies. Understanding the function and regulation of CBX4 could offer insights in devising novel treatments for these malignancies.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
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In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
Chromatin was prepared from human NCCIT cells according to the Abcam Dual-X-ChIP protocol. Cells were fixed with 1.5 mM EGS for 30mins and then 1% formaldehyde for 10min.
The ChIP was performed with 25 μg of chromatin, 2 μg of ab242149 (red), or 2 μg of rabbit normal IgG Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 (gray) and 20 μl of Protein A/G sepharose beads. The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci).
False colour image of Western blot: Anti-CBX4 antibody [EPR23053-7] - ChIP Grade staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab242149 was shown to bind specifically to CBX4. A band was observed at 75 kDa in wild-type HEK-293 cell lysates with no signal observed at this size in Cbx4 knockout cell line Human CBX4 knockout HEK-293 cell line ab261723 (knockout cell lysate ab261666). The band observed in the knockout lysate lane below 75 kDa is likely to represent a truncated form of CBX4. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and Cbx4 knockout HEK-293 cell lysates were analysed.First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3% milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.
All lanes: Western blot - Anti-CBX4 antibody [EPR23053-7] - ChIP Grade (ab242149) at 1/1000 dilution
Lane 1: Wild-type HEK-293 cell lysate at 20 µg
Lane 2: Cbx4 knockout HEK-293 cell lysate at 20 µg
Lane 3: RAW 264.7 cell lysate at 20 µg
Lane 4: HUVEC cell lysate at 20 µg
Performed under reducing conditions.
False colour image of Western blot: Anti-CBX4 antibody [EPR23053-7] - ChIP Grade staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab242149 was shown to bind specifically to CBX4. A band was observed at 75 kDa in wild-type HEK-293 cell lysates with no signal observed at this size in Cbx4 CRISPR-Cas9 edited cell line Human CBX4 knockout HEK-293 cell line ab261723 (CRISPR-Cas9 edited cell lysate ab261666). The band observed in the CRISPR-Cas9 edited lysate lane below 75 kDa is likely to represent a truncated form of CBX4. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and Cbx4 CRISPR-Cas9 edited HEK-293 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3% milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.
All lanes: Western blot - Anti-CBX4 antibody [EPR23053-7] - ChIP Grade (ab242149) at 1/1000 dilution
Lane 1: Wild-type HEK-293 cell lysate at 20 µg
Lane 2: Cbx4 CRISPR-Cas9 edited HEK-293 cell lysate at 20 µg
Lane 3: RAW 264.7 cell lysate at 20 µg
Lane 4: HUVEC cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 61 kDa
Observed band size: 75 kDa
CBX4 was immunoprecipitated from 0.35 mg HCT 116 (Human colorectal carcinoma epithelial cell) whole cell lysate with ab242149 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab242149 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: HCT 116 (Human colorectal carcinoma epithelial cell) whole cell lysate 10ug
Lane 2: ab242149 IP in HCT 116 whole cell lysate
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab242149 in HCT 116 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 15 seconds.
All lanes: Immunoprecipitation - Anti-CBX4 antibody [EPR23053-7] - ChIP Grade (ab242149)
Predicted band size: 52 kDa, 61 kDa
Observed band size: 45 kDa, 60 kDa, 66 kDa, 74 kDa
Immunohistochemical analysis of paraffin-embedded rat hippocampus tissue labeling CBX4 with ab242149 at 1/200 dilution (2.785 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer). Nuclear staining on rat hippocampus (PMID: 28283560). The section was incubated with ab242149 for 30 at RT. The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure time: 26 seconds.
The molecular weight observed is consistent with what has been described in the literature (PMID: 27864346).
All lanes: Western blot - Anti-CBX4 antibody [EPR23053-7] - ChIP Grade (ab242149) at 1/1000 dilution
Lane 1: HCT 116 (human colorectal carcinoma epithelial cell), whole cell lysate at 20 µg
Lane 2: RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate at 20 µg
Lane 3: C6 (rat glial tumor glial cell), whole cell lysate at 20 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 61 kDa
Observed band size: 74 kDa
Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized NCCIT (Human pluripotent embryonic carcinoma epithelial cell) cells labelling CBX4 with ab242149 at 1/600 dilution (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.
Immunohistochemical analysis of paraffin-embedded Mouse hippocampus tissue labeling CBX4 with ab242149 at 1/200 dilution (2.785 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer). Nuclear staining on mouse hippocampus (PMID: 28283560). The section was incubated with ab242149 for 30 at RT. The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling CBX4 with ab242149 at 1/200 dilution (2.785 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer). Nuclear staining on mouse cerebrum (PMID: 28283560). The section was incubated with ab242149 for 30 mins at RT. The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.
The molecular weight observed is consistent with what has been described in the literature (PMID: 27864346).
All lanes: Western blot - Anti-CBX4 antibody [EPR23053-7] - ChIP Grade (ab242149) at 1/1000 dilution
Lane 1: HDLM-2 (human Hodgkin lymphoma) whole cell lysate at 20 µg
Lane 2: NCCIT (human pluripotent embryonic carcinoma epithelial cell), whole cell lysate at 20 µg
Lane 3: Mouse thymus tissue lysate at 20 µg
Lane 4: Rat thymus tissue lysate at 20 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 61 kDa
Observed band size: 74 kDa
Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized HCT 116 (Human colorectal carcinoma epithelial cell) cells labelling CBX4 with ab242149 at 1/600 dilution (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.
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