Anti-KAT1 / HAT1 抗体 [EPR18775]

• IHC-P

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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.

• IHC-P

Supplier Data

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.

• ICC/IF

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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.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-KAT1 / HAT1 antibody [EPR18775] (AB194296)

Intracellular 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.

• IP

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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.

All lanes:

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

Predicted band size: 49 kDa

Observed band size: 45 kDa

false

• ICC/IF

Supplier Data

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.

• IHC-P

Supplier Data

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.

• IHC-P

Supplier Data

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.

• IP

Supplier Data

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.

All lanes:

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

Predicted band size: 49 kDa

Observed band size: 45 kDa

false

• WB

Supplier Data

Western blot - Anti-KAT1 / HAT1 antibody [EPR18775] (AB194296)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

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

Lane 1:

Human fetal liver lysate at 10 µg

Lane 2:

Human fetal kidney lysate at 10 µg

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

false

Exposure time: 5s

• WB

Lab

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.

All lanes:

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

Predicted band size: 49 kDa

false

• WB

Supplier Data

Western blot - Anti-KAT1 / HAT1 antibody [EPR18775] (AB194296)

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

All lanes:

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

Lane 1:

Mouse colon lysate at 20 µg

Lane 2:

Rat colon lysate at 20 µg

Lane 3:

HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 4:

MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 5:

F9 (Mouse embyro testicular cancer cell line) whole cell lysate at 20 µg

Lane 6:

LLC1 (Mouse lung carcinoma cell line) whole cell lysate at 20 µg

Lane 7:

Mouse thymus lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 49 kDa

Observed band size: 45 kDa

false

• WB

Supplier Data

Western blot - Anti-KAT1 / HAT1 antibody [EPR18775] (AB194296)

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.

All lanes:

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

Lane 1:

Mouse brain lysate at 10 µg

Lane 2:

Mouse kidney lysate at 10 µg

Lane 3:

Mouse spleen lysate at 10 µg

Lane 4:

Rat brain lysate at 10 µg

Lane 5:

Rat kidney lysate at 10 µg

Lane 6:

Rat spleen lysate at 10 µg

Lane 7:

C6 (Rat glial tumor cells) whole cell lysate at 10 µg

Lane 8:

RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg

Lane 9:

PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate at 10 µg

Lane 10:

NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 49 kDa

Observed band size: 45 kDa

false