Anti-Lactate Dehydrogenase 抗体 [RM1039] (ab300637)

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

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

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. 

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. 

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.

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.

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 (9.44 μg/ml), followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488, Green) preadsorbed at 1:1000 (2 μg/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 (2.5 μg/ml). Nuclear counter satin is DAPI. 

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 (9.44 μg/ml), followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488, Green) preadsorbed at 1:1000 (2 μg/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 (2.5 μg/ml). Nuclear counter satin is DAPI. 

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.

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.

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.

Lane 1: HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate 10 µg (Input)

Lane 2: ab300637 IP in HeLa whole cell lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab300367 in HeLa whole cell lysate

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

Exposure time: 50 seconds.  

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.

Lane 1: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate 10 µg (Input)

Lane 2: ab300637 IP in NIH/3T3 whole cell lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab300367 in NIH3T3 whole cell lysate

Observed mass (kDa): 37

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

Exposure time: 10 seconds.