Anti-Lactate Dehydrogenase B antibody [60H11]
| Name | Anti-Lactate Dehydrogenase B antibody [60H11] |
|---|---|
| Supplier | Abcam |
| Catalog | ab85319 |
| Prices | $388.00 |
| Sizes | 100 µl |
| Host | Mouse |
| Clonality | Monoclonal |
| Isotype | IgG2b |
| Clone | 60H11 |
| Applications | ICC/IF ICC/IF FC WB IP ELISA IHC-P |
| Species Reactivities | Mouse, Rat, Human |
| Antigen | His tagged recombinant Human lactate dehydrogenase B protein, purified from E |
| Description | Mouse Monoclonal |
| Gene | LDHB |
| Conjugate | Unconjugated |
| Supplier Page | Shop |
Product images
![All lanes : Anti-Lactate Dehydrogenase B antibody [60H11] (ab85319) at 1/2000 dilutionLane 1 : HeLa cell lysateLane 2 : RD cell lysateLane 3 : Ramos cell lysateLane 4 : A431 cell lysateLane 5 : Jurkat cell lysateLane 6 : K562 cell lysateLane 7 : C6 cell lysate](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_3/14561_lactate-dehydrogenase-B-Primary-antibodies-ab85319-1.jpg)
All lanes : Anti-Lactate Dehydrogenase B antibody [60H11] (ab85319) at 1/2000 dilutionLane 1 : HeLa cell lysateLane 2 : RD cell lysateLane 3 : Ramos cell lysateLane 4 : A431 cell lysateLane 5 : Jurkat cell lysateLane 6 : K562 cell lysateLane 7 : C6 cell lysate
ICC/IF image of ab85319 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 (ab85319, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse 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.
IHC image of ab85319 staining in Human breast cancer formalin fixed paraffin embedded tissue section, performed on a Leica BondTM 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 ab85319, 5µg/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.For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
![Overlay histogram showing HeLa cells stained with ab85319 (red line). The cells were fixed with 80% 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 (ab85319, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_3/14564_Lactate-Dehydrogenase-B-Primary-antibodies-ab85319-4.jpg)
Overlay histogram showing HeLa cells stained with ab85319 (red line). The cells were fixed with 80% 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 (ab85319, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
![Anti-Lactate Dehydrogenase B antibody [60H11] (ab85319) at 1/5000 dilution + Active human Lactate Dehydrogenase B full length protein (ab78521) at 0.1 µgSecondaryGoat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.Exposure time : 30 seconds](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_3/14565_Lactate-Dehydrogenase-B-Proteins-and-Peptides-ab78521-1.jpg)
Anti-Lactate Dehydrogenase B antibody [60H11] (ab85319) at 1/5000 dilution + Active human Lactate Dehydrogenase B full length protein (ab78521) at 0.1 µgSecondaryGoat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.Exposure time : 30 seconds
lactate dehydrogenase B was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Mouse monoclonal to Lactate Dehydrogenase B 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 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab85319.Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution.Band: 37kDa: lactate dehydrogenase B
Product References
Ovariectomy induces a shift in fuel availability and metabolism in the - Ovariectomy induces a shift in fuel availability and metabolism in the
Ding F, Yao J, Zhao L, Mao Z, Chen S, Brinton RD. PLoS One. 2013;8(3):e59825.
Quinoline 3-sulfonamides inhibit lactate dehydrogenase A and reverse aerobic - Quinoline 3-sulfonamides inhibit lactate dehydrogenase A and reverse aerobic
Billiard J, Dennison JB, Briand J, Annan RS, Chai D, Colon M, Dodson CS, Gilbert SA, Greshock J, Jing J, Lu H, McSurdy-Freed JE, Orband-Miller LA, Mills GB, Quinn CJ, Schneck JL, Scott GF, Shaw AN, Waitt GM, Wooster RF, Duffy KJ. Cancer Metab. 2013 Sep 6;1(1):19.