重组Anti-GFPT2抗体[EPR19095]

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-GFPT2 antibody [EPR19095] (AB190966)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) cells labeling GFPT2 with ab190966 at 1/150 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue).

Goat anti-Rabbit IgG (Alexa Fluor^® 488) at 1/500 dilution was used as the secondary antibody.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-GFPT2 antibody [EPR19095] (AB190966)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) cells labeling GFPT2 with ab190966 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing cytoplasm staining on U-87 MG cell line.

The nuclear counterstain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).

The negative controls are as follows : -

-ve control 1 : ab190966 at 1/500 dilution followed by ab150120 at 1/1000 dilution.

-ve control 2 : ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.

• IP

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Immunoprecipitation - Anti-GFPT2 antibody [EPR19095] (AB190966)

GFPT2 was immunoprecipitated from 1mg of NIH/3T3 (Mouse embryo fibroblast cells) whole cell lysate with ab190966 at 1/50 dilution.

Western blot was performed from the immunoprecipitate using ab190966 at 1/1000 dilution.

VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1 : NIH/3T3 whole cell lysate 10ug (Input).

Lane 2 : ab190966 IP in NIH/3T3 whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab190966 in NIH/3T3 whole cell lysate.

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

Exposure time : 5 seconds.

All lanes:

Immunoprecipitation - Anti-GFPT2 antibody [EPR19095] (ab190966)

Predicted band size: 77 kDa

Observed band size: 77 kDa

false

• WB

Supplier Data

Western blot - Anti-GFPT2 antibody [EPR19095] (AB190966)

Blocking/Dilution buffer : 5% NFDM/TBST.
Exposure time : Lane 1 : 1 second; Lane 2 : 3 minutes.

Human GFPT2 recombinant protein fragment contains aa2-305 with a His-Tag®, Human GFPT1 recombinant protein fragment contains aa2-288 with a His-Tag®.

All lanes:

Western blot - Anti-GFPT2 antibody [EPR19095] (ab190966) at 1/2000 dilution

Lane 1:

Human GFPT2 recombinant protein fragment at 0.01 µg

Lane 2:

Human GFPT1 recombinant protein fragment at 0.01 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: 77 kDa

Observed band size: 33 kDa

false

• WB

Supplier Data

Western blot - Anti-GFPT2 antibody [EPR19095] (AB190966)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-GFPT2 antibody [EPR19095] (ab190966) at 1/2000 dilution

Lane 1:

U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) whole cell lysate at 10 µg

Lane 2:

Human glioma lysate at 10 µg

Secondary

Lane 1:

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

Lane 2:

Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 77 kDa

Observed band size: 77 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-GFPT2 antibody [EPR19095] (AB190966)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-GFPT2 antibody [EPR19095] (ab190966) at 1/2000 dilution

Lane 1:

Mouse heart lysate at 10 µg

Lane 2:

Mouse kidney lysate at 10 µg

Lane 3:

Mouse spleen lysate at 10 µg

Lane 4:

Rat heart lysate at 10 µg

Lane 5:

Rat spleen lysate at 10 µg

Lane 6:

NIH/3T3 (Mouse embryo fibroblast cell line) 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: 77 kDa

Observed band size: 77 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-GFPT2 antibody [EPR19095] (AB190966)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time :

Lane 1 & 2 : 3 minutes.

Lane 3 & 4 : 5 seconds.

All lanes:

Western blot - Anti-GFPT2 antibody [EPR19095] (ab190966) at 1/2000 dilution

Lane 1:

Human placenta lysate at 20 µg

Lane 2:

Mouse placenta lysate at 20 µg

Lane 3:

A-375 (Human malignant melanoma cell line) whole cell lysate at 20 µg

Lane 4:

Rat placenta 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: 77 kDa

Observed band size: 77 kDa

false