Anti-LPCAT1 antibody [EPR19882]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LPCAT1 antibody [EPR19882] (AB214034)

Immunohistochemical analysis of paraffin-embedded human lung tissue labeling LPCAT1 with ab214034 at 1/2000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution.

Cytoplasmic or granular staining on the type two alveolar epithelial cells of human lung 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) 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-LPCAT1 antibody [EPR19882] (AB214034)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed A431 (Human epidermoid carcinoma cell line) cells labeling LPCAT1 with ab214034 at 1/50 dilution (red) compared with aRabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730; black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody; blue). Goat Anti-Rabbit IgG (Alexa Fluor^® 488) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LPCAT1 antibody [EPR19882] (AB214034)

Immunohistochemical analysis of paraffin-embedded human liver cancer tissue labeling LPCAT1 with ab214034 at 1/2000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution.

Cytoplasmic or granular staining on the liver cancer cells which contain abundant lipid drops 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) 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-LPCAT1 antibody [EPR19882] (AB214034)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed A549 (Human lung carcinoma cell line) cells labeling LPCAT1 with ab214034 at 1/50 dilution (red) compared withRabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730; black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody; blue). Goat Anti-Rabbit IgG (Alexa Fluor^® 488) at 1/2000 dilution was used as the secondary antibody.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-LPCAT1 antibody [EPR19882] (AB214034)

Immunofluorescent analysis of 4% paraformaldehyde-fixed 0.1% Triton X-100 permeabilized A431 (Human epidermoid carcinoma cell line) cells labeling LPCAT1 with ab214034 at 1/250 dilution followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on A431 cells. The nuclear counterstain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution (red).

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-LPCAT1 antibody [EPR19882] (AB214034)

Immunofluorescent analysis of 4% paraformaldehyde-fixed 0.1% Triton X-100 permeabilized A549 (Human lung carcinoma cell line) cells labeling LPCAT1 with ab214034 at 1/250 dilution followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on A549 cells. The nuclear counterstain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution (red).

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LPCAT1 antibody [EPR19882] (AB214034)

Immunohistochemical analysis of paraffin-embedded human lung cancer tissue labeling LPCAT1 with ab214034 at 1/2000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution.

Cytoplasmic or granular staining on the lung cancer cells 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) at 1/500 dilution.

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

• IP

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Immunoprecipitation - Anti-LPCAT1 antibody [EPR19882] (AB214034)

LPCAT1 was immunoprecipitated from 0.35 mg of A549 (Human lung carcinoma cell line) whole cell lysate with ab214034 at 1/30 dilution.

Western blot was performed from the immunoprecipitate using ab214034 at 1/1000 dilution.

VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution

Lane 1 : A549 whole cell lysate 10μg (Input).

Lane 2 : ab214034 IP in A549 whole cell lysate.

Lane 3 : Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab214034 in A549 whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 1 second.

All lanes:

Immunoprecipitation - Anti-LPCAT1 antibody [EPR19882] (ab214034)

Predicted band size: 59 kDa

false

• WB

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Western blot - Anti-LPCAT1 antibody [EPR19882] (AB214034)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : Lane 1-4/8 : 4 seconds; Lane 5/6 : 8 seconds; Lane 7 : 3 minutes.

Lanes 1 - 6:

Western blot - Anti-LPCAT1 antibody [EPR19882] (ab214034) at 1/2000 dilution

Lanes 7 - 8:

Western blot - Anti-LPCAT1 antibody [EPR19882] (ab214034) at 1/10000 dilution

Lane 1:

Human fetal brain lysate at 10 µg

Lane 2:

Human testis lysate at 10 µg

Lane 3:

A431 (Human epidermoid carcinoma cell line) whole cell lysate at 10 µg

Lane 4:

A549 (Human lung carcinoma cell line) whole cell lysate at 10 µg

Lane 5:

Human fetal spleen lysate at 10 µg

Lane 6:

Human fetal kidney lysate at 10 µg

Lane 7:

Human prostate cancer lysate at 10 µg

Lane 8:

PC-3 (Human prostate adenocarcinoma cell line) whole cell lysate at 10 µg

Secondary

All lanes:

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

Predicted band size: 59 kDa

Observed band size: 59 kDa

false