Anti-Lactate Dehydrogenase antibody [EP1566Y]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lactate Dehydrogenase antibody [EP1566Y] (AB52488)

ab52488 staining Lactate Dehydrogenase in human breast carcinoma tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/2000. ab97051 was used as the secondary antibody.

Negative control 1 : PBS in place of primary antibody.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Lactate Dehydrogenase antibody [EP1566Y] (AB52488)

ab52488 staining Lactate Dehydrogenase in HeLa (human cervix adenocarcinoma) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at a dilution of 1/100. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a dilution of 1/1000. ab7291 and ab150120 were used as counterstains for primary antibody ab52488 and secondary antibody ab150077 respectively and DAPI was used as a nuclear counterstain.

Negative control 1 : Rabbit primary antibody and anti-mouse secondary antibody (ab150120)
Negative control 2 : Mouse primary antibody (ab7291) and anti-rabbit secondary antibody (ab150077)

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Lactate Dehydrogenase antibody [EP1566Y] (AB52488)

ICC/IF image of unpurified ab52488 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab52488, 1μg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor^® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43μM.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Lactate Dehydrogenase antibody [EP1566Y] (AB52488)

Overlay histogram showing HeLa cells stained with unpurified ab52488 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab52488, 1/50 dilution) for 30 min at 22°C. The secondary antibody used was DyLight^® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (1μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a slightly decreased signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

• IP

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Immunoprecipitation - Anti-Lactate Dehydrogenase antibody [EP1566Y] (AB52488)

ab52488 immunoprecipitating Lactate Dehydrogenase. 10μg of cell lysate was incubated with primary antibody at a dilution of 1/30 and VeriBlot for IP Detection Reagent (HRP) (ab131366) at a dilution of 1/10000.

Lane 1 : HeLa (human cervix adenocarcinoma) whole cell lysate (10ug)
Lane 2 : HeLa (human cervix adenocarcinoma) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab52488 in HeLa (human cervix adenocarcinoma) whole cell lysate

All lanes:

Immunoprecipitation - Anti-Lactate Dehydrogenase antibody [EP1566Y] (ab52488)

Predicted band size: 37 kDa

Observed band size: 36 kDa

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• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Lactate Dehydrogenase antibody [EP1566Y] (AB52488)

Intracellular Flow Cytometry analysis of Raw264.7 (mouse abelson murine leukemia virus-induced tumor) whole cell lysate labelling Lactate Dehydrogenase with purified ab52488 at 1/190 (red). Cells were fixed with 4% paraformaldehyde. Alexa Fluorr^® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lactate Dehydrogenase antibody [EP1566Y] (AB52488)

ab52488 staining Lactate Dehydrogenase in mouse liver tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/2000. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

Negative control 1 : PBS in place of primary antibody.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lactate Dehydrogenase antibody [EP1566Y] (AB52488)

ab52488 staining Lactate Dehydrogenase in mouse liver tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/2000. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

Negative control 1 : PBS in place of primary antibody.

• WB

Lab

Western blot - Anti-Lactate Dehydrogenase antibody [EP1566Y] (AB52488)

All lanes:

Western blot - Anti-Lactate Dehydrogenase antibody [EP1566Y] (ab52488) at 1/5000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma) whole cell lysate at 10 µg

Lane 2:

HEK293 (human embryonic kidney) whole cell lysate at 10 µg

Lane 3:

Human heart tissue 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: 37 kDa

Observed band size: 36 kDa

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• WB

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Western blot - Anti-Lactate Dehydrogenase antibody [EP1566Y] (AB52488)

Blocking and diluting buffer 5% NFDM/TBST

All lanes:

Western blot - Anti-Lactate Dehydrogenase antibody [EP1566Y] (ab52488) at 1/20000 dilution

Lane 1:

Mouse kidney tissue lysate at 10 µg

Lane 2:

Rat kidney tissue 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 1/20000 dilution

Predicted band size: 37 kDa

Observed band size: 36 kDa

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• WB

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Western blot - Anti-Lactate Dehydrogenase antibody [EP1566Y] (AB52488)

All lanes:

Western blot - Anti-Lactate Dehydrogenase antibody [EP1566Y] (ab52488) at 1/100000 dilution

All lanes:

Hela cell lysate at 10 µg

Secondary

All lanes:

Goat anti-rabbit HRP labeled at 1/2000 dilution

Predicted band size: 37 kDa

Observed band size: 37 kDa

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