This polyclonal antibody was produced against rat immunoglobulin κ light chain peptide (SVTDQDSKDSTYSMSSTLSL)1,2
It cross-react with some other species (see immunoblot in figure1). It can be used western blot and dot blot.
Western blot of serum of variaus species with the rabbit anti-immunoglobulin κ light
chain polyclonal antibody 1: Rat; 2: Mouse; 3:Marmoset. In the species, Only a band at <30k (neighboring 24k ) is detected.
Procedure for western blot of sera with rabbit anti-immunoglobulin κ light chain polyclonal antibodies
- Rat or mouse sea
- Extracting buffer solutions (100 mM Tris-HCl, pH 8.8, 2% SDS, 20% sucrose, 0.06% BPB, 7M urea, 2M thiourea and 0.1M DTT)
- SDS-PAGE gel (10-20% gradient or 12.5% isocratically)
- PVDF or nitrocellulose membrane
- Blocking buffer (BSA or skim milk etc)
- RESVO rabbit anti- immunoglobulin κ light chain polyclonal antibody
- HRP conjugated anti-rabbit IgG
- ECL western blot substrate
- Sera mixed at a volume ratio of 1:1 with protein extracting buffer solutions(Vortex more than 15 min).
- The mixture was subjected to SDS-PAGE gel .
- Electrophoresed proteins were transferred from the SDS-PAGE gel to a PVDF (or nitrocellulose) membrane.
- The membrane is blocked for 1 hr at room temperature in a blocking buffer solution (Including BSA or skim milk etc).
- The membrane was then incubated overnight at 4°C in a blocking buffer solution containing 1:500-2000 diluted anti-immunoglobulin
κ light chain polyclonal antibody and finally transferred into a blocking buffer solution containing a HRP conjugated anti-rabbit IgG.
- Chemiluminescence from HRP elicited by ECL western blot substrate was detected by western blot imager.
Rat or mouse serum samples should diluted from 10-100 times with PBS etc, because rodent serum include high concentration of
immunoglobulin κ light chain.