Anti-PFKM antibody [EPR10734(B)]

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-PFKM antibody [EPR10734(B)] (AB154804)

Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling PFKM with purified ab154804 at 1/300 dilution (10μg/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor^® 488) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-PFKM antibody [EPR10734(B)] (AB154804)

Immunocytochemistry/ Immunofluorescence analysis of SH-SY5Y (Human neuroblastoma epithelial cell) cells labeling PFKM with Purified ab154804 at 1 : 300 dilution (8.7μg/ml). Cells were fixed in 100% Methanol and cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1 : 200 (2.5 μg/ml). ab150077 Goat anti rabbit IgG(Alexa Fluor^® 488) was used as the secondary antibody at 1 : 1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-PFKM antibody [EPR10734(B)] (AB154804)

Intracellular flow cytometric analysis of permeabilized SHSY5Y cells labeling PFKM with ab154804 at 1/100 dilution (red) or a rabbit IgG (green - negative control).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-PFKM antibody [EPR10734(B)] (AB154804)

Immunofluorescence analysis of HeLa cells labeling PFKM with ab154804 at 1/50 dilution.

• WB

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Western blot - Anti-PFKM antibody [EPR10734(B)] (AB154804)

All lanes:

Western blot - Anti-PFKM antibody [EPR10734(B)] (ab154804) at 1/1000 dilution

Lane 1:

Fetal brain lysate at 10 µg

Lane 2:

Fetal heart lysate at 10 µg

Lane 3:

Ramos lysate at 10 µg

Lane 4:

SHSY5Y lysate at 10 µg

Lane 5:

HeLa lysate at 10 µg

Predicted band size: 85 kDa

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• WB

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Western blot - Anti-PFKM antibody [EPR10734(B)] (AB154804)

Blocking and diluting buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-PFKM antibody [EPR10734(B)] (ab154804) at 1/2000 dilution

All lanes:

Human fetal heart lysates at 15 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 85 kDa

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• WB

Unknown

Western blot - Anti-PFKM antibody [EPR10734(B)] (AB154804)

Blocking and diluting buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-PFKM antibody [EPR10734(B)] (ab154804) at 1/2000 dilution

Lane 1:

Rat skeletal muscle lysates at 15 µg

Lane 2:

Mouse skeletal muscle lysates at 15 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 1/20000 dilution

Predicted band size: 22 kDa,85 kDa

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• WB

AbReview37428****

Western blot - Anti-PFKM antibody [EPR10734(B)] (AB154804)

Western blot analysis of Pig skeletal muscle tissue lysate using ab154804 (1 : 2000 in 1x PBS + 0.1% Tween-20 for 1 hour at 22°C) to label PFKM. A IRDye® 680-conjugated Donkey anti-rabbit IgG (1 : 40000) was used as the secondary antibody. The image shows increasing amounts of loaded protein for antibody optimization. The far left lane is the molecular weight standard.

All lanes:

Western blot - Anti-PFKM antibody [EPR10734(B)] (ab154804)

Predicted band size: 85 kDa

Observed band size: 81 kDa

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Exposure time: 5min

This image is courtesy of an Abreview submitted by Eric England

• WB

Unknown

Western blot - Anti-PFKM antibody [EPR10734(B)] (AB154804)

Blocking and diluting buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-PFKM antibody [EPR10734(B)] (ab154804) at 1/10000 dilution

All lanes:

Pig heart lysates at 15 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 1/20000 dilution

Predicted band size: 85 kDa

false

• WB

Lab

Western blot - Anti-PFKM antibody [EPR10734(B)] (AB154804)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure times : Lane 1 : 3 seconds; Lane 2 : 10 seconds; Lane 3 : 180 seconds.

All lanes:

Western blot - Anti-PFKM antibody [EPR10734(B)] (ab154804) at 1/1000 dilution

Lane 1:

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

Lane 2:

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 3:

GST tagged Recombinant Human PFKL protein (Full length, 116 KDa)

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 1/20000 dilution

Predicted band size: 85 kDa

Observed band size: 85 kDa

false