Anti-FACL4 antibody [EPR17587-42] - BSA and Azide free

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-FACL4 antibody [EPR17587-42] - BSA and Azide free (AB251419)

This data was developed using ab205199, the same antibody clone in a different buffer formulation.

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling FACL4 with ab205199 at 1/120 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.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-FACL4 antibody [EPR17587-42] - BSA and Azide free (AB251419)

This data was developed using ab205199, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling FACL4 with ab205199 at 1/50 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic 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 : ab205199 at 1/50 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.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-FACL4 antibody [EPR17587-42] - BSA and Azide free (AB251419)

This data was developed using ab205199, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (Human liver hepatocellular carcinoma) cells labeling FACL4 with ab205199 at 1/50 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on HepG2 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 : ab205199 at 1/50 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

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FACL4 antibody [EPR17587-42] - BSA and Azide free (AB251419)

This data was developed using ab205199, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human liver carcinoma tissue labeling FACL4 with ab205199 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Cytoplasmic staining on Human hepatocellular carcinoma tissue 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

Unknown

Immunoprecipitation - Anti-FACL4 antibody [EPR17587-42] - BSA and Azide free (AB251419)

This data was developed using ab205199, the same antibody clone in a different buffer formulation.

FACL4 was immunoprecipitated from 1mg of Mouse brain whole cell lysate with ab205199 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab205199 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

Lane 1 : Mouse brain whole cell lysate 10 μg (Input). Lane 2 : ab205199 IP in Mouse brain whole cell lysate. Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab205199 in Mouse brain whole cell lysate
Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 8 seconds

All lanes:

Immunoprecipitation - Anti-FACL4 antibody [EPR17587-42] (<a href='/en-us/products/primary-antibodies/facl4-antibody-epr17587-42-ab205199'>ab205199</a>)

Predicted band size: 79 kDa

false

• WB

Supplier Data

Western blot - Anti-FACL4 antibody [EPR17587-42] - BSA and Azide free (AB251419)

This data was developed using ab205199, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-FACL4 antibody [EPR17587-42] (<a href='/en-us/products/primary-antibodies/facl4-antibody-epr17587-42-ab205199'>ab205199</a>) at 1/5000 dilution

Lane 1:

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

Lane 2:

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

Secondary

All lanes:

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

Predicted band size: 44 kDa,79 kDa

Observed band size: 24 kDa,44 kDa,72 kDa,79 kDa

false

Exposure time: 30s

• WB

Supplier Data

Western blot - Anti-FACL4 antibody [EPR17587-42] - BSA and Azide free (AB251419)

This data was developed using ab205199, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-FACL4 antibody [EPR17587-42] (<a href='/en-us/products/primary-antibodies/facl4-antibody-epr17587-42-ab205199'>ab205199</a>) at 1/1000 dilution

Lane 1:

Human fetal brain lysate at 10 µg

Lane 2:

Human fetal heart lysate at 10 µg

Lane 3:

Human fetal kidney lysate at 10 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 79 kDa

Observed band size: 79 kDa

false

Exposure time: 1min

• WB

Supplier Data

Western blot - Anti-FACL4 antibody [EPR17587-42] - BSA and Azide free (AB251419)

This data was developed using ab205199, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-FACL4 antibody [EPR17587-42] (<a href='/en-us/products/primary-antibodies/facl4-antibody-epr17587-42-ab205199'>ab205199</a>) at 1/5000 dilution

All lanes:

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: 79 kDa

Observed band size: 79 kDa

false

Exposure time: 3min