Rabbit Recombinant Monoclonal EZHIP antibody. Suitable for IHC-P, Flow Cyt (Intra), WB, ICC/IF and reacts with Human, Mouse samples.
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
IHC-P | Flow Cyt (Intra) | WB | ICC/IF | IP | |
---|---|---|---|---|---|
Human | Tested | Tested | Tested | Tested | Not recommended |
Mouse | Not recommended | Expected | Tested | Expected | Not recommended |
Rat | Not recommended | Not recommended | Not recommended | Not recommended | Not recommended |
Species | Dilution info | Notes |
---|---|---|
Species Human | 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 Mouse, Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/500 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/1000 | Notes - |
Species Mouse | Dilution info 1/1000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/500 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human, Mouse, Rat | Dilution info - | Notes - |
Select an associated product type
Inhibits PRC2/EED-EZH1 and PRC2/EED-EZH2 complex function by inhibiting EZH1/EZH2 methyltransferase activity, thereby causing down-regulation of histone H3 trimethylation on 'Lys-27' (H3K27me3) (PubMed:29909548, PubMed:30923826, PubMed:31086175, PubMed:31451685). Probably inhibits methyltransferase activity by limiting the stimulatory effect of cofactors such as AEBP2 and JARID2 (PubMed:30923826). Inhibits H3K27me3 deposition during spermatogenesis and oogenesis (By similarity).
CXorf67, EZHIP, EZH inhibitory protein
Rabbit Recombinant Monoclonal EZHIP antibody. Suitable for IHC-P, Flow Cyt (Intra), WB, ICC/IF and reacts with Human, Mouse samples.
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
EPR28396-50
Affinity purification Protein A
Unsuitable for mouse IHC-P.
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.
The EZHIP (EZH Inhibitory Protein) also known as CATACOMB is a protein that inhibits the activity of EZH2 an important enzyme in the PRC2 (Polycomb Repressive Complex 2). EZHIP is a protein with a molecular mass of approximately 90 kDa. It expresses in various tissues with notable presence in the male and female reproductive tracts as well as in several cancer cell lines. The protein's role involves modulating chromatin structure by interfering with methyltransferase activity affecting gene silencing.
EZHIP interacts with the Polycomb Repressive Complex 2 (PRC2) altering its methylation activity on histone H3 at lysine 27 (H3K27me3) an important mark for epigenetic gene silencing. By engaging the complex EZHIP hinders the catalytic efficiency of EZH2 reducing the repression of target genes. This interaction significantly impacts epigenetic regulation as alterations to methylation patterns influence cellular functions and differentiation processes.
EZHIP's function affects pathways related to chromatin modification and epigenetic regulation. It is central to the regulation of the JAK-STAT signaling pathway and interacts closely with EZH2 and possibly other members of the Polycomb group proteins. EZHIP shapes the chromatin landscape influencing the transcriptional output by collaborating or competing with other transcription factors and modifiers within these pathways.
EZHIP's aberrant expression is linked to cancers including gliomas and germ cell tumors. The relationship between EZHIP and EZH2 becomes important as alterations in EZH2 activity lead to abnormal gene silencing associated with oncogenesis. Additionally research indicates that dysregulation involving EZHIP might contribute to developmental disorders through improper epigenetic marks. Further study about its interactions could provide insights into therapeutic approaches for conditions linked to epigenetic misregulation.
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.
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.
In Western blot, anti- H3 antibody (Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade ab176842) loading control staining at 1/100000 dilution.
In Western blot, anti-GAPDH antibody (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/200000 dilution.
All lanes: Western blot - Anti-EZHIP antibody [EPR28396-50] (ab313392) at 1/1000 dilution
Lane 1: Mouse testis cytoplasmic fraction at 50 µg
Lane 2: Mouse testis nuclear fraction at 50 µg
All lanes: Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Developed using the ECL technique.
Observed band size: 75 kDa
Exposure time: 180s
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized 293T (human embryonic kidney epithelial cell, Left) / U-2 OS (human bone osteosarcoma epithelial cell, Right) cells labelling EZHIP with ab313392 at 1/500 dilution (0.1 ug)/Red (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) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody. Low expression: 293T (PMID: 34880314).
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized U-2 OS (human bone osteosarcoma epithelial cell) cells labelling CXorf67 with ab313392 at 1/500 (1.022 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing nuclear staining in U-2 OS cell line.Low expression: 293T (PMID: 34880314).Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.
Immunohistochemical analysis of paraffin-embedded Human cardiac muscle tissue labeling CXorf67 with ab313392 at 1/200 (2.555 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Negative control: No staining on human cardiac muscle.The section was incubated with ab313392 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemical analysis of paraffin-embedded Human skeletal muscl tissue labeling CXorf67 with ab313392 at 1/200 (2.555 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Negative control: No staining on human skeletal muscle.The section was incubated with ab313392 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemical analysis of paraffin-embedded Human placenta tissue labeling CXorf67 with ab313392 at 1/200 (2.555 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Positive staining on human placenta.The section was incubated with ab313392 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling CXorf67 with ab313392 at 1/200 (2.555 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Positive staining on human testis (PMID: 31451685).The section was incubated with ab313392 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Blocking and diluting buffer and concentration: 5% NFDM/TBST
Low expression: 293T (PMID: 34880314).
The molecular weight observed is consistent with what has been described in the literature (PMID: 34880314).
Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.
In Western blot, anti-GAPDH antibody (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/200000 dilution.
Exposure time: 92 seconds
All lanes: Western blot - Anti-EZHIP antibody [EPR28396-50] (ab313392) at 1/1000 dilution
Lane 1: U-2OS (human bone osteosarcoma epithelial cell) whole cell lysate at 50 µg
Lane 2: 293T (human embryonic kidney epithelial cell) whole cell lysate at 50 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 50, 75 kDa
Exposure time: 92s
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