Anti-Lactate Dehydrogenase antibody

• IHC-P

AbReview16772****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lactate Dehydrogenase antibody (AB47010)

ab47010 at a 1/400 dilution staining LDHA in human skeletal muscle sections by Immunohistochemistry (formalin/PFA-fixed paraffin-embedded sections) incubated for 20 minutes at 25°C. Heat mediated antigen retrieval used, EDTA ph 9.0, 100°C for 20 minutes. Secondary used undiluted, goat anti-mouse/rabbit IgG conjugated to HRP polymer.

This image is from an anonymous abreview.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Lactate Dehydrogenase antibody (AB47010)

ICC/IF image of ab47010 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab47010, 1μg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. 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). DAPI was used to stain the cell nuclei (blue).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lactate Dehydrogenase antibody (AB47010)

IHC image of Lactate Dehydrogenase staining in human breast carcinoma FFPE section, performed on a Leica Bond^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab47010, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

• IP

Unknown

Immunoprecipitation - Anti-Lactate Dehydrogenase antibody (AB47010)

LDHA was immunoprecipitated using 0.5mg Hela whole cell extract, 5μg of Rabbit polyclonal to LDHA and 50μl of protein G magnetic beads (+). No antibody was added to the control (-).

The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.

Proteins were eluted by addition of 40μl SDS loading buffer and incubated for 10min at 70°C; 10μl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab47010.

Secondary : Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).

Band : 36kDa; LDHA

All lanes:

Immunoprecipitation - Anti-Lactate Dehydrogenase antibody (ab47010)

Predicted band size: 37 kDa

true

Exposure time: 12min

• WB

AbReview26601****

Western blot - Anti-Lactate Dehydrogenase antibody (AB47010)

Blocking step : 5% Milk for 1 hour at 24°C

Primary antibody incubation for 12 hours at 4°C

All lanes:

Western blot - Anti-Lactate Dehydrogenase antibody (ab47010) at 1/500 dilution

All lanes:

Zebrafish whole cell lysate at 10 µg

Secondary

All lanes:

HRP conjugated Goat anti-rabbit monoclonal secondary antibody at 1/5000 dilution

Predicted band size: 37 kDa

Observed band size: 22 kDa,36 kDa

true

Exposure time: 30s

This image is courtesy of an anonymous Abreview

• WB

Project3322****

Western blot - Anti-Lactate Dehydrogenase antibody (AB47010)

All lanes:

LDHA antibody (ab47010) at 1 µg/mL

Lane 1:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 2:

Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate at 10 µg

Lane 3:

A431 (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 4:

HEK293 Human embryonic kidney cell line Whole Cell Lysate at 10 µg

Secondary

All lanes:

IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

Predicted band size: 37 kDa

Observed band size: 36 kDa

false

• WB

Unknown

Western blot - Anti-Lactate Dehydrogenase antibody (AB47010)

All lanes:

Western blot - Anti-Lactate Dehydrogenase antibody (ab47010) at 1 µg/mL

All lanes:

Western blot - Recombinant human Lactate Dehydrogenase protein (Active) (<a href='/en-us/products/proteins-peptides/recombinant-human-lactate-dehydrogenase-protein-active-ab93699'>ab93699</a>) at 0.1 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-preadsorbed-ab97080'>ab97080</a>) at 1/5000 dilution

Predicted band size: 37 kDa

true

Exposure time: 10s

• WB

Supplier Data

Western blot - Anti-Lactate Dehydrogenase antibody (AB47010)

Observed bands

Lane 1 : 40 kDa

Lane 2 : 37 kDa

Lane 3 : 62 kDa

Lane 4 : 37 kDa

All lanes:

Western blot - Anti-Lactate Dehydrogenase antibody (ab47010)

Lane 1:

LDHA recombinant

Lane 2:

LDHB recombinant

Lane 3:

LDHC recombinant

Lane 4:

HeLa

Predicted band size: 37 kDa

false

Exposure time: 30s