Anti-PFKM + PFKP antibody [EPR17314] - BSA and Azide free

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-PFKM + PFKP antibody [EPR17314] - BSA and Azide free (AB242413)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling PFKP with ab204131 at 1/250 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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab204131)

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-PFKM + PFKP antibody [EPR17314] - BSA and Azide free (AB242413)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed MCF7 (Human breast adenocarcinoma cell line) cells labeling PFKP with ab204131 at 1/250 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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab204131)

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-PFKM + PFKP antibody [EPR17314] - BSA and Azide free (AB242413)

Immunofluorescent analysis of 100% methanl-fixed, 0.1% Triton X-100 permeabilized MCF7 (Human breast adenocarcinoma cell line) cells labeling PFKP with ab204131 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on MCF7 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody at 1/1000 dilution (red).
The negative controls are as follows :
-ve control 1 : ab204131 at 1/250 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody at 1/1000 dilution.
-ve control 2 : Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab204131)

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-PFKM + PFKP antibody [EPR17314] - BSA and Azide free (AB242413)

Immunofluorescent analysis of 100% methanl-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling PFKP with ab204131 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (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 Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody at 1/1000 dilution (red).
The negative controls are as follows :
-ve control 1 : ab204131 at 1/250 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody at 1/1000 dilution.
-ve control 2 : Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab204131)

• IP

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Immunoprecipitation - Anti-PFKM + PFKP antibody [EPR17314] - BSA and Azide free (AB242413)

PFKP was immunoprecipitated from 1mg of MCF7 (Human breast adenocarcinoma cell line) whole cell lysate with ab204131 at 1/80 dilution.

Western blot was performed from the immunoprecipitate using ab204131 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 : MCF7 whole cell lysate 10ug (Input).

Lane 2 : ab204131 IP in MCF7 whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab204131 in MCF7 whole cell lysate.

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

Exposure time : 10 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab204131)

All lanes:

Immunoprecipitation - Anti-PFKM + PFKP antibody [EPR17314] (<a href='/en-us/products/primary-antibodies/pfkm-pfkp-antibody-epr17314-ab204131'>ab204131</a>)

false

• WB

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Western blot - Anti-PFKM + PFKP antibody [EPR17314] - BSA and Azide free (AB242413)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab204131).

Blocking/diluting buffer and concentration : 5% NFDM/TBST

Exposure time : Lane1 : 10 seconds
Lane 2 : 1 seconds

All lanes:

Western blot - Anti-PFKM + PFKP antibody [EPR17314] (<a href='/en-us/products/primary-antibodies/pfkm-pfkp-antibody-epr17314-ab204131'>ab204131</a>) at 1/1000 dilution

Lane 1:

Western blot - Recombinant Human PFKP protein (GST tag N-Terminus) (<a href='/en-us/products/proteins-peptides/recombinant-human-pfkp-protein-ab132823'>ab132823</a>)

Lane 2:

DDK tagged Recombinant Human PFKM protein (Full length, 85 KDa)

Secondary

Lanes 1 - 2:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/20000 dilution

Lanes 1 - 2:

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 1/20000 dilution

false