![Lactate Dehydrogenase B was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal 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 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 ab75167.Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).Band: 34kDa; Lactate Dehydrogenase B](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_3/14545_ab75167-197831-IPV023ab751673min.jpg)
Lactate Dehydrogenase B was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal 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 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 ab75167.Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).Band: 34kDa; Lactate Dehydrogenase B
ab75167 at 5 µg/ml staining lactate dehydrogenase B in Human placenta by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).
Anti-Lactate Dehydrogenase B antibody (ab75167) at 1 µg/ml + fetal muscle lysate at 10 µgSecondaryHRP conjugated anti-Rabbit IgG at 1/50000 dilution
IHC image of ab75167 staining in human normal liver (bile duct cells) 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 ab75167, 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.
ab75167 at 5 µg/ml staining lactate dehydrogenase B in Human placenta by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).
All lanes : Anti-Lactate Dehydrogenase B antibody (ab75167) at 1 µg/mlLane 1 : MarkerLane 2 : Zebrafish brain homogenate at 20 µgLane 3 : Zebrafish heart homogenate at 20 µgLane 4 : Zebrafish liver homogenate at 20 µgLane 5 : Zebrafish skeletal muscle homogenate at 20 µgSecondaryGoat polyclonal to Rabbit IgG – H&L – Pre-Adsorbed (HRP) at 1/6000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.
ICC/IF image of ab75167 stained DU145 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 ab75167 at 1µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) 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.
Application: IHC. Species+tissue/cell type: Human lung adenocarcinoma cell line A549. Primary antibody dilution: 1:100. Secondary antibody: Goat anti-rabbit AlexaFluor 488. Secondary antibody dilution: 1:400
