- SynonymRPL,Protein L
- SourceBiotinylated Recombinant Protein L, His,Avitag™, also known as Recombinant Peptostreptococcus magnus Protein L, was expressed in E.Coli at ACRObiosystems.Predicted N-terminus: MetRequest for sequence
- Molecular Characterization
This protein carries a polyhistidine tag at the N-terminus, followed by an Avi tag (Avitag™). The protein has a calculated MW of 43.7 kDa. The protein migrates as 50 kDa under reducing (R) condition (SDS-PAGE). This protein can bind to VL-Kappa.
- BiotinylationBiotinylation of this product is performed using Avitag™ technology. Briefly, the single lysine residue in the Avitag is enzymatically labeled with biotin.
- Biotin:Protein RatioThe biotin to protein ratio is 0.5-1 as determined by the HABA assay.
- EndotoxinLess than 1.0 EU per μg by the LAL method.
- Purity
>95% as determined by SDS-PAGE.
>90% as determined by SEC-MALS.
- Formulation
Lyophilized from 0.22 μm filtered solution in PBS, pH7.4. Normally trehalose is added as protectant before lyophilization.
Contact us for customized product form or formulation.
- Reconstitution
Please see Certificate of Analysis for specific instructions.
For best performance, we strongly recommend you to follow the reconstitution protocol provided in the CoA.
- Storage
For long term storage, the product should be stored at lyophilized state at -20°C or lower.
Please protect from light and avoid repeated freeze-thaw cycles.
This product is stable after storage at:
- -20°C to -70°C for 12 months in lyophilized state;
- -70°C for 3 months under sterile conditions after reconstitution.
Biotinylated Recombinant Protein L, His,Avitag™ on SDS-PAGE under reducing (R) condition. The gel was stained overnight with Coomassie Blue. The purity of the protein is greater than 95%.
The purity of Biotinylated Recombinant Protein L, His,Avitag™ (Cat. No. RPL-P81Q7) was more than 90% and the molecular weight of this protein is around 85-100 kDa verified by SEC-MALS.
Immobilized Mouse Human chimeric Anti-BCMA mAb (Aa01, Mouse IgG1) at 2 μg/mL (100 μL/well) can bind Biotinylated Recombinant Protein L, His,Avitag (Cat. No. RPL-P81Q7) with a linear range of 0.2-2 ng/mL (QC tested).
Immobilized Ipilimumab at 2 μg/mL (100 μL/well) can bind Biotinylated Recombinant Protein L, His,Avitag (Cat. No. RPL-P81Q7) with a linear range of 0.2-2 ng/mL (Routinely tested).
Immobilized nivolumab at 2 μg/mL (100 μL/well) can bind Biotinylated Recombinant Protein L, His,Avitag (Cat. No. RPL-P81Q7) with a linear range of 0.2-2 ng/mL (Routinely tested).
Loaded Biotinylated Recombinant Protein L, His,Avitag™(Cat. No. RPL-P81Q7) on SA Biosensor, can bind Monoclonal Anti-Human CD19 Antibody, Mouse IgG2a with an affinity constant of 13.6 nM as determined in BLI assay (ForteBio Octet Red96e) (Routinely tested).
Loaded Biotinylated Recombinant Protein L, His,Avitag™(Cat. No. RPL-P81Q7) on SA Biosensor, can bind Anti-CD19 (FMC63), scFv, His Tag with an affinity constant of 74.3 nM as determined in BLI assay (ForteBio Octet Red96e) (Routinely tested).
Loaded Biotinylated Recombinant Protein L, His,Avitag™(Cat. No. RPL-P81Q7) on SA Biosensor, can bind Anti-BCMA scFv,His Tag with an affinity constant of 6.56 nM as determined in BLI assay (ForteBio Octet Red96e) (Routinely tested).
Loaded Biotinylated Recombinant Protein L, His,Avitag™(Cat. No. RPL-P81Q7) on SA Biosensor, can bindAnti-Human BCMA MAb (human IgG1) with an affinity constant of 14 nM as determined in BLI assay (ForteBio Octet Red96e) (Routinely tested).
2e5 of anti-BCMA CAR-293 cells were stained with 100 μL of 10 μg/mL of Biotinylated Recombinant Protein L, His,Avitag™(Cat. No. RPL-P81Q7) and negative control protein respectively, washed and then followed by PE-SA and analyzed with FACS (QC tested).
2e5 of anti-CD19 CAR-293 cells were stained with 100 μL of 10 μg/mL of Biotinylated Recombinant Protein L, His,Avitag™(Cat. No. RPL-P81Q7) and negative control protein respectively, washed and then followed by PE-SA and analyzed with FACS (QC tested).
- BackgroundProtein L was isolated from the surface of bacterial species Peptostreptococcus magnus and was found to bind Ig(IgG,IgM,IgA,IgE and IgD) through L chain interaction, from which the name was suggested. Despite this wide-ranging binding capability with respect to Ig classes, Protein L is not a universal immunoglobilin-binding protein. Binding of Protein L to immunoglobulins is restricted to those containing kappa light chains (i.e., k chain of the VL domain). In humans and mice, kappa (k) light chains predominate. The remaining immunoglobulins have lambda (l) light chains. The recombinant protein contains four immunoglobulin (Ig) binding domains (Bdomains) of the native protein. Besides antibody, protein L is also suitable for binding of a wide range of antibody fragments such as Fabs, single-chain variable fragments (scFv), and domain antibodies (Dabs).
- References
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