![Lysozyme was immunoprecipitated using 0.5mg HL60 whole cell extract, 5µg of Rabbit polyclonal to Lysozyme 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, HL60 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 ab2408.Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).Band: 16kDa; Lysozyme](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_3/18184_ab2408-171834-IPV018ab240820mMod.jpg)
Lysozyme was immunoprecipitated using 0.5mg HL60 whole cell extract, 5µg of Rabbit polyclonal to Lysozyme 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, HL60 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 ab2408.Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).Band: 16kDa; Lysozyme
Ab2408 staining lysozyme in human colon. Staining is localised to cytoplasm.Left panel: with primary antibody at 2 ug/ml. Right panel: isotype control.Sections were stained using an automated system (DAKO Autostainer Plus), at room temperature: sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
All lanes : Anti-Lysozyme antibody (ab2408) at 1 µg/mlLane 1 : HL60 (Human promyelocytic leukemia cell line) Whole Cell Lysate Lane 2 : Spleen (Human) Tissue Lysate - adult normal tissue (ab29699)Lysates/proteins at 10 µg per lane.SecondaryGoat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.
Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human tonsil tissue, staining Lysozyme with ab2408.