Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-KAT1 / HAT1 antibody [EPR18775] (ab194296)

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Overview

  • Product name

    Anti-KAT1 / HAT1 antibody [EPR18775]
    See all KAT1 / HAT1 primary antibodies

  • Description

    Rabbit monoclonal [EPR18775] to KAT1 / HAT1

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt, IHC-P, WB, ICC/IF, IPmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment within Human KAT1/ HAT1 aa 200 to the C-terminus. The exact sequence is proprietary.
    Database link: O14929

  • Positive control

    • WB: HeLa, MCF7, F9, LLC1, C6, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates; Human fetal liver and fetal kidney lysates; Mouse thymus lysate; Mouse and rat colon, brain, kidney and spleen lysates. IHC-P: Human tonsil, Human gastric adenocarcinoma, mouse colon and rat colon tissues. ICC/IF: HeLa and F9 cells. Flow Cyt: HeLa cells. IP: HeLa and F9 whole cell lysates.

  • General 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 rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab194296 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Flow Cyt 1/150.
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/1000. Detects a band of approximately 45 kDa (predicted molecular weight: 49 kDa).
ICC/IF 1/500.
IP 1/50.

Target

  • Function

    Acetylates soluble but not nucleosomal histone H4 at 'Lys-5' (H4K5ac) and 'Lys-12' (H4K12ac) and, to a lesser extent, acetylates histone H2A at 'Lys-5' (H2AK5ac). Has intrinsic substrate specificity that modifies lysine in recognition sequence GXGKXG. May be involved in nucleosome assembly during DNA replication and repair as part of the histone H3.1 and H3.3 complexes. May play a role in DNA repair in response to free radical damage.

  • Sequence similarities

    Belongs to the HAT1 family.

  • Developmental stage

    Highly expressed in mitotic cells (at protein level).

  • Cellular localization

    Nucleus matrix and Cytoplasm. Nucleus. Nucleus matrix. Nucleus > nucleoplasm. Localization is predominantly nuclear in normal cells. Treatment with hydrogen peroxide or ionizing radiation enhances nuclear localization through redistribution of existing protein.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • HAT 1 antibody
    • hat1 antibody
    • HAT1_HUMAN antibody
    • Histidine aminotransferase 1 antibody
    • Histone acetyltransferase 1 antibody
    • Histone acetyltransferase 1 antibody
    • Histone acetyltransferase type B catalytic subunit antibody
    • KAT1 antibody
    see all

Images

  • Western blot - Anti-KAT1 / HAT1 antibody [EPR18775] (ab194296)
    Western blot - Anti-KAT1 / HAT1 antibody [EPR18775] (ab194296)

    Lane 1: Wild-type HAP1 cell lysate (40 µg)
    Lane 2: KAT1/HAT1 knockout HAP1 cell lysate (40 µg)
    Lane 3: HeLa cell lysate (20 µg)
    Lane 4:Jurkat cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab194296 observed at 48 kDa. Red - loading control, ab18058, observed at 37 kDa.

    ab194296 was shown to recognize KAT1/HAT1 in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when KAT1/HAT1 knockout samples were examined. Wild-type and KAT1 / HAT1 knockout samples were subjected to SDS-PAGE. Ab194296 and ab18058 (loading control to GAPDH) were diluted at 1/1000 and 1/10,000 dilution respectively and incubated overnight at 4oC. Blots were developed with IRDye® 800CW Goat anti-Rabbit IgG (H + L) and IRDye® 680 Goat anti-Mouse IgG (H + L) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-KAT1 / HAT1 antibody [EPR18775] (ab194296)
    Western blot - Anti-KAT1 / HAT1 antibody [EPR18775] (ab194296)
    All lanes : Anti-KAT1 / HAT1 antibody [EPR18775] (ab194296) at 1/1000 dilution

    Lane 1 : Human fetal liver lysate
    Lane 2 : Human fetal kidney lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/10000 dilution

    Predicted band size: 49 kDa
    Observed band size: 45 kDa
    why is the actual band size different from the predicted?


    Exposure time: 5 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-KAT1 / HAT1 antibody [EPR18775] (ab194296)
    Western blot - Anti-KAT1 / HAT1 antibody [EPR18775] (ab194296)
    All lanes : Anti-KAT1 / HAT1 antibody [EPR18775] (ab194296) at 1/1000 dilution

    Lane 1 : Mouse colon lysate
    Lane 2 : Rat colon lysate
    Lane 3 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
    Lane 4 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
    Lane 5 : F9 (Mouse embyro testicular cancer cell line) whole cell lysate
    Lane 6 : LLC1 (Mouse lung carcinoma cell line) whole cell lysate
    Lane 7 : Mouse thymus lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 49 kDa
    Observed band size: 45 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1 and 2: 3 minutes; Lane 3 and 4: 2 seconds; Lane 5, 6 and 7: 5 seconds

  • Western blot - Anti-KAT1 / HAT1 antibody [EPR18775] (ab194296)
    Western blot - Anti-KAT1 / HAT1 antibody [EPR18775] (ab194296)
    All lanes : Anti-KAT1 / HAT1 antibody [EPR18775] (ab194296) at 1/1000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Mouse kidney lysate
    Lane 3 : Mouse spleen lysate
    Lane 4 : Rat brain lysate
    Lane 5 : Rat kidney lysate
    Lane 6 : Rat spleen lysate
    Lane 7 : C6 (Rat glial tumor cells) whole cell lysate
    Lane 8 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 9 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
    Lane 10 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 49 kDa
    Observed band size: 45 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1, 2, 4, 5 and 6: 3 minutes; Lane 3: 1 minute; Lane 7, 8, 9 and 10: 5 seconds.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KAT1 / HAT1 antibody [EPR18775] (ab194296)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KAT1 / HAT1 antibody [EPR18775] (ab194296)

    Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling KAT1 / HAT1 with ab194296 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus staining on the germinal center of Human tonsil is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KAT1 / HAT1 antibody [EPR18775] (ab194296)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KAT1 / HAT1 antibody [EPR18775] (ab194296)

    Immunohistochemical analysis of paraffin-embedded Human gastric adenocarcinoma tissue labeling KAT1 / HAT1 with ab194296 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus staining on cancer cells of Human gastric adenocarcinoma is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KAT1 / HAT1 antibody [EPR18775] (ab194296)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KAT1 / HAT1 antibody [EPR18775] (ab194296)

    Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling KAT1 / HAT1 with ab194296 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus staining on epithelial cells of mouse colon is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KAT1 / HAT1 antibody [EPR18775] (ab194296)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KAT1 / HAT1 antibody [EPR18775] (ab194296)

    Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling KAT1 / HAT1 with ab194296 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus staining on epithelial cells of rat colon is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-KAT1 / HAT1 antibody [EPR18775] (ab194296)
    Immunocytochemistry/ Immunofluorescence - Anti-KAT1 / HAT1 antibody [EPR18775] (ab194296)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling KAT1 / HAT1 with ab194296 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on HeLa cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
    The negative controls are as follows:
    -ve control 1: ab194296 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

  • Immunocytochemistry/ Immunofluorescence - Anti-KAT1 / HAT1 antibody [EPR18775] (ab194296)
    Immunocytochemistry/ Immunofluorescence - Anti-KAT1 / HAT1 antibody [EPR18775] (ab194296)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized F9 (Mouse embyro testicular cancer cell line) cells labeling KAT1 / HAT1 with ab194296 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on F9 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
    The negative controls are as follows:
    -ve control 1: ab194296 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

  • Flow Cytometry - Anti-KAT1 / HAT1 antibody [EPR18775] (ab194296)
    Flow Cytometry - Anti-KAT1 / HAT1 antibody [EPR18775] (ab194296)

    Flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling KAT1 / HAT1 with ab194296 at 1/150 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/500 dilution was used as the secondary antibody.

  • Immunoprecipitation - Anti-KAT1 / HAT1 antibody [EPR18775] (ab194296)
    Immunoprecipitation - Anti-KAT1 / HAT1 antibody [EPR18775] (ab194296)

    KAT1 / HAT1 was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate with ab194296 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab194296 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: HeLa whole cell lysate 10ug (Input). 

    Lane 2: ab194296 IP in HeLa whole cell lysate. 

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab194296 in HeLa whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.
    Exposure time: 30 seconds.

  • Immunoprecipitation - Anti-KAT1 / HAT1 antibody [EPR18775] (ab194296)
    Immunoprecipitation - Anti-KAT1 / HAT1 antibody [EPR18775] (ab194296)

    KAT1 / HAT1 was immunoprecipitated from 1mg of F9 (Mouse embyro testicular cancer cell line) whole cell lysate with ab194296 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab194296 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: F9 whole cell lysate 10ug (Input). 

    Lane 2: ab194296 IP in F9 whole cell lysate. 

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab194296 in F9 whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.
    Exposure time: 30 seconds.

References

This product has been referenced in:

  • Fan P  et al. Overexpressed histone acetyltransferase 1 regulates cancer immunity by increasing programmed death-ligand 1 expression in pancreatic cancer. J Exp Clin Cancer Res 38:47 (2019). Read more (PubMed: 30709380) »
  • Aksu M  et al. Xpo7 is a broad-spectrum exportin and a nuclear import receptor. J Cell Biol 217:2329-2340 (2018). Read more (PubMed: 29748336) »
See all 2 Publications for this product

Publishing research using ab194296? Please let us know so that we can cite the reference in this datasheet.

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