Anti-ATE1 antibody [EPR13667(2)] - N-terminal

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATE1 antibody [EPR13667(2)] - N-terminal (AB199423)

Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma tissue labeling ATE1 with ab199423 at 1/250 followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500. Cytoplasmic staining on human hepatocellular carcinoma tissue is observed (Subcellular location : Nucleus and cytoplasm [UniProt]). Counter-stained with hematoxylin.

Negative 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.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-ATE1 antibody [EPR13667(2)] - N-terminal (AB199423)

Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling ATE1 with ab199423 at 1/220 dilution (red) compared with a rabbit monoclonal IgG isotype control (black) and a unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATE1 antibody [EPR13667(2)] - N-terminal (AB199423)

Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling ATE1 with ab199423 at 1/250 followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500. Cytoplasm staining on Rat kidney tissue is observed (Subcellular location : Nucleus and Cytoplasm [UniProt]) . Counter stained with Hematoxylin.
Negative 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-ATE1 antibody [EPR13667(2)] - N-terminal (AB199423)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling ATE1 with ab199423 at 1/250 followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500. Cytoplasm staining on Mouse liver tissue is observed (Subcellular location : Nucleus and Cytoplasm [UniProt]) . Counter stained with Hematoxylin.
Negative 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.

• WB

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Western blot - Anti-ATE1 antibody [EPR13667(2)] - N-terminal (AB199423)

Blocking and diluting buffer 5% NFDM/TBST

All lanes:

Western blot - Anti-ATE1 antibody [EPR13667(2)] - N-terminal (ab199423) at 1/10000 dilution

Lane 1:

HepG2 (Human liver hepatocellular carcinoma) whole cell lysate at 20 µg

Lane 2:

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

Lane 3:

293 (Human epithelial cells from embryonic kidney) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 59 kDa

Observed band size: 59 kDa

true

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-ATE1 antibody [EPR13667(2)] - N-terminal (AB199423)

Blocking and diluting buffer 5% NFDM/TBST

All lanes:

Western blot - Anti-ATE1 antibody [EPR13667(2)] - N-terminal (ab199423) at 1/10000 dilution

All lanes:

Human fetal liver tissue lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 59 kDa

Observed band size: 59 kDa

true

Exposure time: 10s

• WB

Supplier Data

Western blot - Anti-ATE1 antibody [EPR13667(2)] - N-terminal (AB199423)

Blocking and diluting buffer 5% NFDM/TBST

All lanes:

Western blot - Anti-ATE1 antibody [EPR13667(2)] - N-terminal (ab199423) at 1/1000 dilution

Lane 1:

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

Lane 2:

Raw264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg

Lane 3:

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

Lane 4:

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

Lane 5:

Mouse kidney tissue lysate at 10 µg

Lane 6:

Rat kidney tissue lysate at 10 µg

Lane 7:

Rat spleen tissue lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 59 kDa

Observed band size: 59 kDa

true

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-ATE1 antibody [EPR13667(2)] - N-terminal (AB199423)

Blocking and diluting buffer 5% NFDM/TBST

All lanes:

Western blot - Anti-ATE1 antibody [EPR13667(2)] - N-terminal (ab199423) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 10 µg

Lane 2:

Mouse spleen tissue lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 59 kDa

Observed band size: 59 kDa

true

Exposure time: 1min

• WB

CiteAb

Western blot - Anti-ATE1 antibody [EPR13667(2)] - N-terminal (AB199423)

ATE1 western blot using anti-ATE1 antibody [EPR13667(2)] - N-terminal ab199423. Publication image and figure legend from Singh, K., Gupta, A., et al., 2020, Sci Rep, PubMed 31953451.

ab199423 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab199423 please see the product overview.

ATE1 expression is upregulated by hypertrophic stimuli. (A) Quantitative real-time PCR analysis of mRNA levels of ATE1 in Ang II treated H9C2 cells. (B) Transcriptional levels of ATE1 in heart samples from rat subjected to ligation of right renal artery (Ligated) and sham-operated control (Sham) rats. (C) Western blot analysis of ATE1 protein levels in heart samples from sham and renal ligated rats. Data were derived from experiments performed in triplicate and normalized to GAPDH content. Statistical analysis was carried out by student’s two-tailed t-test (*shows non-specific binding of antibody).

false