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Rabbit Recombinant Monoclonal EZHIP antibody. Suitable for IHC-P, Flow Cyt (Intra), WB, ICC/IF and reacts with Human, Mouse samples.

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Images

Western blot - Anti-EZHIP antibody [EPR28396-50] (AB313392), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-EZHIP antibody [EPR28396-50] (AB313392), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-EZHIP antibody [EPR28396-50] (AB313392), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EZHIP antibody [EPR28396-50] (AB313392), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EZHIP antibody [EPR28396-50] (AB313392), expandable thumbnail

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IHC-PFlow Cyt (Intra)WBICC/IFIP
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

Tested
Tested

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.

Not recommended
Not recommended

Species

Mouse, Rat

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1/500

Notes

-

Expected
Expected

Species

Mouse

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Rat

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1/1000

Notes

-

Species

Mouse

Dilution info

1/1000

Notes

-

Not recommended
Not recommended

Species

Rat

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1/500

Notes

-

Expected
Expected

Species

Mouse

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Rat

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Human, Mouse, Rat

Dilution info

-

Notes

-

Associated Products

Select an associated product type

1 product for Alternative Version

Target data

Function

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).

Alternative names

Recommended products

Rabbit Recombinant Monoclonal EZHIP antibody. Suitable for IHC-P, Flow Cyt (Intra), WB, ICC/IF and reacts with Human, Mouse samples.

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

EPR28396-50

Purification technique

Affinity purification Protein A

Specificity

Unsuitable for mouse IHC-P.

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

Notes

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:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

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.

Biological function summary

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.

Pathways

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.

Associated diseases and disorders

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.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

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.

8 product images

  • Western blot - Anti-EZHIP antibody [EPR28396-50] (ab313392), expandable thumbnail

    Western blot - Anti-EZHIP antibody [EPR28396-50] (ab313392)

    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

    Secondary

    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 Cytometry (Intracellular) - Anti-EZHIP antibody [EPR28396-50] (ab313392), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-EZHIP antibody [EPR28396-50] (ab313392)

    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).

  • Immunocytochemistry/ Immunofluorescence - Anti-EZHIP antibody [EPR28396-50] (ab313392), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-EZHIP antibody [EPR28396-50] (ab313392)

    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EZHIP antibody [EPR28396-50] (ab313392), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EZHIP antibody [EPR28396-50] (ab313392)

    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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EZHIP antibody [EPR28396-50] (ab313392), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EZHIP antibody [EPR28396-50] (ab313392)

    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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EZHIP antibody [EPR28396-50] (ab313392), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EZHIP antibody [EPR28396-50] (ab313392)

    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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EZHIP antibody [EPR28396-50] (ab313392), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EZHIP antibody [EPR28396-50] (ab313392)

    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

  • Western blot - Anti-EZHIP antibody [EPR28396-50] (ab313392), expandable thumbnail

    Western blot - Anti-EZHIP antibody [EPR28396-50] (ab313392)

    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

    Secondary

    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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