Anti-Lactate Dehydrogenase antibody [RM1039]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lactate Dehydrogenase antibody [RM1039] (AB300637)

Immunohistochemical analysis of paraffin-embedded human tonsil tissue labelling lactate dehydrogenase A, B, C with ab300637 at 1/5000 (0.094 µg/ml) followed by a ready to use LeicaDS9800 (BOND^™ Polymer Refine Detection).

Positive staining on human tonsil is observed.

The section was incubated with ab300637 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

Counterstained with Hematoxylin.

Secondary antibody only control : PBS was used instead of primary antibody followed by ready to use secondary antibody LeicaDS9800 (BOND^™ Polymer Refine Detection kit).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins was used.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Lactate Dehydrogenase antibody [RM1039] (AB300637)

Immunofluorescent analysis of 4% paraformaldehyde fixed and 0.1% TritonX-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labeling lactate dehydrogenase A, B, C with ab300637 at 1/50 dilution (9.44µg/ml), followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488, Green) preadsorbed at 1 : 1000 (2µg/mL).

Confocal image showing cytoplasmic and weak nuclear staining in HeLa cell line.

ab195889 anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) (Red) was used as a counterstain at 1 : 200 dilution (2.5µg/ml).

Nuclear counter satin is DAPI.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Lactate Dehydrogenase antibody [RM1039] (AB300637)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed and 90% methanol permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling lactate dehydrogenase A, B, C with ab300637 at 1/500 dilution (0.1µg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lactate Dehydrogenase antibody [RM1039] (AB300637)

Immunohistochemical analysis of paraffin-embedded human colon carcionoma tissue labelling Lactate Dehydrogenase A, B, C with ab300637 at 1/5000 (0.094 µg/ml) followed by a ready to use LeicaDS9800 (BOND^™ Polymer Refine Detection).

Positive staining on human colon carcinoma is observed.

The section was incubated with ab300637 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

Counterstained with Hematoxylin.

Secondary antibody only control : PBS was used instead of primary antibody followed by ready to use secondary antibody LeicaDS9800 (BOND^™ Polymer Refine Detection kit).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins was used.

• IP

Lab

Immunoprecipitation - Anti-Lactate Dehydrogenase antibody [RM1039] (AB300637)

Lactate Dehydrogenase was immunoprecipitated from 0.35 mg HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate with ab300637 at 1/30 dilution (2 µg in 0.35 mg lysates). Western blot was performed on the immunoprecipitate using ab300637 at 1/5000 dilution.

VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

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

All lanes:

Immunoprecipitation - Anti-Lactate Dehydrogenase antibody [RM1039] (ab300637) at 1/5000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 10 µg

Lane 2:

ab300637 at 1/30 IP in HeLa whole cell lysate at 10 µg

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab300637 in HeLa whole cell lysate at 10 µg

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 50s

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Lactate Dehydrogenase antibody [RM1039] (AB300637)

Immunofluorescent analysis of 4% paraformaldehyde fixed and 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labeling lactate dehydrogenase A, B, C with ab300637 at 1/50 dilution (9.44µg/ml), followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488, Green) preadsorbed at 1 : 1000 (2µg/mL).

Confocal image showing cytoplasmic and weak nuclear staining in NIH/3T3 cell line.

ab195889 anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) (Red) was used as a counterstain at 1 : 200 dilution (2.5µg/ml).

Nuclear counter satin is DAPI.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lactate Dehydrogenase antibody [RM1039] (AB300637)

Immunohistochemical analysis of paraffin-embedded rat cardiac muscle tissue labelling Lactate Dehydrogenase A, B, C with ab300637 at 1/5000 (0.094 µg/ml) followed by a ready to use LeicaDS9800 (BOND^™ Polymer Refine Detection).

Positive staining on rat cardiac muscle is observed.

The section was incubated with ab300637 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

Counterstained with Hematoxylin.

Secondary antibody only control : PBS was used instead of primary antibody followed by ready to use secondary antibody LeicaDS9800 (BOND^™ Polymer Refine Detection kit).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins was used.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Lactate Dehydrogenase antibody [RM1039] (AB300637)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed and 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling lactate dehydrogenase A, B, C with ab300637 at 1/500 dilution (0.1µg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lactate Dehydrogenase antibody [RM1039] (AB300637)

Immunohistochemical analysis of paraffin-embedded mouse cardiac muscle tissue labelling Lactate Dehydrogenase A, B, C with ab300637 at 1/5000 (0.094 µg/ml) followed by a ready to use LeicaDS9800 (BOND^™ Polymer Refine Detection).

Positive staining on mouse cardiac muscle is observed.

The section was incubated with ab300637 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

Counterstained with Hematoxylin.

Secondary antibody only control : PBS was used instead of primary antibody followed by ready to use secondary antibody LeicaDS9800 (BOND^™ Polymer Refine Detection kit).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins was used.

• IP

Lab

Immunoprecipitation - Anti-Lactate Dehydrogenase antibody [RM1039] (AB300637)

Lactate Dehydrogenase was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast) whole cell lysate with ab300637 at 1/30 dilution (2 µg in 0.35 mg lysates). Western blot was performed on the immunoprecipitate using ab300637 at 1/1000 dilution.

VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

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

All lanes:

Immunoprecipitation - Anti-Lactate Dehydrogenase antibody [RM1039] (ab300637) at 1/1000 dilution

Lane 1:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 10 µg

Lane 2:

ab300637 at 1/30 IP in NIH/3T3 whole cell lysate at 10 µg

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab300637 in NIH/3T3 whole cell lysate at 10 µg

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 10s

• WB

Lab

Western blot - Anti-Lactate Dehydrogenase antibody [RM1039] (AB300637)

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

All lanes:

Western blot - Anti-Lactate Dehydrogenase antibody [RM1039] (ab300637) at 1/1000 dilution

Lane 1:

Human heart lysate at 20 µg

Lane 2:

Mouse heart lysate at 20 µg

Lane 3:

Mouse skeletal muscle lysate at 20 µg

Lane 4:

Rat heart lysate at 20 µg

Lane 5:

Rat skeletal muscle lysate at 20 µg

Lane 6:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 7:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 8:

C6 (rat glial tumor glial cell) whole cell lysate at 20 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 1/100000 dilution

Predicted band size: 37 kDa

Observed band size: 35-37 kDa

false

Exposure time: 10s

• WB

Lab

Western blot - Anti-Lactate Dehydrogenase antibody [RM1039] (AB300637)

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

All lanes:

Western blot - Anti-Lactate Dehydrogenase antibody [RM1039] (ab300637) at 1/1000 dilution

Lane 1:

LDH-A recombinant protein containing his-tag (2-332 aa) at 10 ng

Lane 2:

LDH-B recombinant protein containing his-tag (1-334 aa)  at 10 ng

Lane 3:

LDH-C recombinant protein containing GST-tag (1-332 aa)  at 10 ng

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 1/100000 dilution

Observed band size: 37 kDa,62 kDa

false

Exposure time: 80s