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

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. This information is proprietary to Abcam and/or its suppliers.
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
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 production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR18775
Isotype

IgG
Research areas

Alternative Versions
Assay kits
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Conjugation kits
- PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
- APC Conjugation Kit - Lightning-Link® (ab201807)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Recombinant Protein
- Recombinant Human KAT1 / HAT1 protein (ab93687)
Related Products
- VeriBlot for IP Detection Reagent (HRP) (ab131366)

The Abpromise guarantee

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.

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.

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.
Target information above from: UniProt accession O14929 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 8520 Human
- Entrez Gene: 107435 Mouse
- Entrez Gene: 296501 Rat
- Omim: 603053 Human
- SwissProt: O14929 Human
- SwissProt: Q8BY71 Mouse
- SwissProt: Q5M939 Rat
- Unigene: 632532 Human
- Unigene: 272472 Mouse
- Unigene: 139751 Rat
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

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 4^oC. 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)

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)

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)

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)

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)

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)

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)

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)

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)

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 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)

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)

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

Flow cytometry protocols
Immunoprecipitation protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

Datasheet download

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

ab194296 has been referenced in 8 publications.

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). PubMed: 30709380
Gruber JJ et al. HAT1 Coordinates Histone Production and Acetylation via H4 Promoter Binding. Mol Cell 75:711-724.e5 (2019). PubMed: 31278053
Loktev SS & Ogneva IV DNA Methylation of Mouse Testes, Cardiac and Lung Tissue During Long-Term Microgravity Simulation. Sci Rep 9:7974 (2019). PubMed: 31138883
Gruber JJ et al. Chromatin Remodeling in Response to BRCA2-Crisis. Cell Rep 28:2182-2193.e6 (2019). PubMed: 31433991
Yang G et al. HAT1 signaling confers to assembly and epigenetic regulation of HBV cccDNA minichromosome. Theranostics 9:7345-7358 (2019). PubMed: 31695772
Xia P et al. MicroRNA-377 exerts a potent suppressive role in osteosarcoma through the involvement of the histone acetyltransferase 1-mediated Wnt axis. J Cell Physiol 234:22787-22798 (2019). PubMed: 31152456
Hunt LC et al. A Key Role for the Ubiquitin Ligase UBR4 in Myofiber Hypertrophy in Drosophila and Mice. Cell Rep 28:1268-1281.e6 (2019). PubMed: 31365869
Aksu M et al. Xpo7 is a broad-spectrum exportin and a nuclear import receptor. J Cell Biol 217:2329-2340 (2018). PubMed: 29748336

Key features and details

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Assay kits

Compatible Secondaries

Conjugation kits

Isotype control

Recombinant Protein

Related Products

Applications

The Abpromise guarantee

Target

Function

Sequence similarities

Developmental stage

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

Datasheet download

Certificate of Compliance

References (8)

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