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

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Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EZHIP antibody [EPR28396-24] - BSA and Azide free (AB316857), expandable thumbnail
  • Western blot - Anti-EZHIP antibody [EPR28396-24] - BSA and Azide free (AB316857), expandable thumbnail
  • Western blot - Anti-EZHIP antibody [EPR28396-24] - BSA and Azide free (AB316857), expandable thumbnail
  • Immunoprecipitation - Anti-EZHIP antibody [EPR28396-24] - BSA and Azide free (AB316857), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-EZHIP antibody [EPR28396-24] - BSA and Azide free (AB316857), expandable thumbnail

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Constituents: 100% PBS

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-PICC/IFWBFlow Cyt (Intra)IP
Human
Tested
Tested
Tested
Tested
Tested
Mouse
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended
Rat
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended

Tested
Tested

Species

Human

Dilution info

-

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

-

Notes

-

Not recommended
Not recommended

Species

Mouse, Rat

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Mouse, Rat

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Mouse, Rat

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Mouse, Rat

Dilution info

-

Notes

-

Associated Products

Select an associated product type

1 products for Alternative Version

    Target data

    Alternative names

    EZH inhibitory protein, CXorf67, EZHIP

    Recommended products

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

    Alternative names

    EZH inhibitory protein, CXorf67, EZHIP

    Key facts

    Isotype

    IgG

    Form

    Liquid

    Clonality

    Monoclonal

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

    Yes

    Clone number

    EPR28396-24

    Purification technique

    Affinity purification Protein A

    Concentration
    Loading...

    Storage

    Shipped at conditions

    Blue Ice

    Appropriate long-term storage conditions

    +4°C

    Notes

    ab316857 is the carrirer-free version of Anti-EZHIP antibody [EPR28396-24] ab316856.

    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.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    Supplementary info

    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.

    9 product images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EZHIP antibody [EPR28396-24] - BSA and Azide free (ab316857), expandable thumbnail

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EZHIP antibody [EPR28396-24] - BSA and Azide free (ab316857)

      This data was developed using Anti-EZHIP antibody [EPR28396-24] ab316856, the same antibody clone in a different buffer formulation.

      Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling EZHIP with Anti-EZHIP antibody [EPR28396-24] ab316856 at 1/500 (0.996 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

      Negative control: No staining on human liver.

      The section was incubated with Anti-EZHIP antibody [EPR28396-24] ab316856 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

      Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

    • Western blot - Anti-EZHIP antibody [EPR28396-24] - BSA and Azide free (ab316857), expandable thumbnail

      Western blot - Anti-EZHIP antibody [EPR28396-24] - BSA and Azide free (ab316857)

      This data was developed using Anti-EZHIP antibody [EPR28396-24] ab316856, the same antibody clone in a different buffer formulation.

      Negative control: A204.

      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 [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.

      All lanes: Western blot - Anti-EZHIP antibody [EPR28396-24] (Anti-EZHIP antibody [EPR28396-24] ab316856) at 1/1000 dilution

      Lane 1: U-2OS (human bone osteosarcoma epithelial cell) whole cell lysate at 50 µg

      Lane 2: A-204 (human muscle rhabdomyosarcoma 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 1/20000 dilution

      Observed band size: 40 kDa, 75 kDa, 36 kDa

      Exposure time: 10s

      Negative control: A204.

      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 [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.

    • Western blot - Anti-EZHIP antibody [EPR28396-24] - BSA and Azide free (ab316857), expandable thumbnail

      Western blot - Anti-EZHIP antibody [EPR28396-24] - BSA and Azide free (ab316857)

      This data was developed using Anti-EZHIP antibody [EPR28396-24] ab316856, the same antibody clone in a different buffer formulation.

      Low expression: 293T (PMID: 34880314).

      The molecular weight observed is consistent with what has been described in the literature (PMID: 34880314).

      In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.

      All lanes: Western blot - Anti-EZHIP antibody [EPR28396-24] (Anti-EZHIP antibody [EPR28396-24] ab316856) 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: Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

      Observed band size: 40 kDa, 75 kDa, 36 kDa

      Exposure time: 6s

    • Immunoprecipitation - Anti-EZHIP antibody [EPR28396-24] - BSA and Azide free (ab316857), expandable thumbnail

      Immunoprecipitation - Anti-EZHIP antibody [EPR28396-24] - BSA and Azide free (ab316857)

      This data was developed using Anti-EZHIP antibody [EPR28396-24] ab316856, the same antibody clone in a different buffer formulation.

      EZHIP was immunoprecipitated from 0.35 mg U-2 OS (human bone osteosarcoma epithelial cell) whole cell lysate with Anti-EZHIP antibody [EPR28396-24] ab316856 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using Anti-EZHIP antibody [EPR28396-24] ab316856 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.

      Lane 1: U-2 OS (human bone osteosarcoma epithelial cell) whole cell lysate

      Lane 2: Anti-EZHIP antibody [EPR28396-24] ab316856 IP in U-2 OS (human bone osteosarcoma epithelial cell) whole cell lysate

      Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-EZHIP antibody [EPR28396-24] ab316856 in U-2 OS whole cell lysate

      Lysate was freshly made and used for Western blotting immediately to minimize protein degradation.

      All lanes: Immunoprecipitation - Anti-EZHIP antibody [EPR28396-24] (Anti-EZHIP antibody [EPR28396-24] ab316856) at 1/30 dilution

      All lanes: U-2 OS (human bone osteosarcoma epithelial cell) whole cell lysate

      Secondary

      All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution

      Exposure time: 10s

      EZHIP was immunoprecipitated from 0.35 mg U-2 OS (human bone osteosarcoma epithelial cell) whole cell lysate with Anti-EZHIP antibody [EPR28396-24] ab316856 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using Anti-EZHIP antibody [EPR28396-24] ab316856 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.

      Lane 1: U-2 OS (human bone osteosarcoma epithelial cell) whole cell lysate

      Lane 2: Anti-EZHIP antibody [EPR28396-24] ab316856 IP in U-2 OS (human bone osteosarcoma epithelial cell) whole cell lysate

      Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-EZHIP antibody [EPR28396-24] ab316856 in U-2 OS whole cell lysate

      Lysate was freshly made and used for Western blotting immediately to minimize protein degradation.

    • Flow Cytometry (Intracellular) - Anti-EZHIP antibody [EPR28396-24] - BSA and Azide free (ab316857), expandable thumbnail

      Flow Cytometry (Intracellular) - Anti-EZHIP antibody [EPR28396-24] - BSA and Azide free (ab316857)

      This data was developed using Anti-EZHIP antibody [EPR28396-24] ab316856, the same antibody clone in a different buffer formulation.

      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 Anti-EZHIP antibody [EPR28396-24] ab316856 at 1/500 dilution (0.1 ug)/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).

      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.

    • Immunocytochemistry/ Immunofluorescence - Anti-EZHIP antibody [EPR28396-24] - BSA and Azide free (ab316857), expandable thumbnail

      Immunocytochemistry/ Immunofluorescence - Anti-EZHIP antibody [EPR28396-24] - BSA and Azide free (ab316857)

      This data was developed using Anti-EZHIP antibody [EPR28396-24] ab316856, the same antibody clone in a different buffer formulation.

      Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized U-2 OS (human bone osteosarcoma epithelial cell) cells labelling EZHIP with Anti-EZHIP antibody [EPR28396-24] ab316856 at 1/100 (4.98 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 [EPR28396-24] - BSA and Azide free (ab316857), expandable thumbnail

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EZHIP antibody [EPR28396-24] - BSA and Azide free (ab316857)

      This data was developed using Anti-EZHIP antibody [EPR28396-24] ab316856, the same antibody clone in a different buffer formulation.

      Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling EZHIP with Anti-EZHIP antibody [EPR28396-24] ab316856 at 1/500 (0.996 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

      Negative control: No staining on human kidney (PMID: 31451685).

      The section was incubated with Anti-EZHIP antibody [EPR28396-24] ab316856 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

      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 [EPR28396-24] - BSA and Azide free (ab316857), expandable thumbnail

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EZHIP antibody [EPR28396-24] - BSA and Azide free (ab316857)

      This data was developed using Anti-EZHIP antibody [EPR28396-24] ab316856, the same antibody clone in a different buffer formulation.

      Immunohistochemical analysis of paraffin-embedded Human placenta tissue labeling EZHIP with Anti-EZHIP antibody [EPR28396-24] ab316856 at 1/500 (0.996 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

      Positive staining on human placenta (PMID: 27699219).

      The section was incubated with Anti-EZHIP antibody [EPR28396-24] ab316856 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

      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 [EPR28396-24] - BSA and Azide free (ab316857), expandable thumbnail

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EZHIP antibody [EPR28396-24] - BSA and Azide free (ab316857)

      This data was developed using Anti-EZHIP antibody [EPR28396-24] ab316856, the same antibody clone in a different buffer formulation.

      Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling EZHIP with Anti-EZHIP antibody [EPR28396-24] ab316856 at 1/500 (0.996 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

      Positive staining on human testis (PMID: 31451685).

      The section was incubated with Anti-EZHIP antibody [EPR28396-24] ab316856 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

      Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

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