膜杰作
Star Staining
产品描述（Product Details）
| Assay Type | Sandwich-ELISA |
| Analyte | Protein L |
| Format | 96T |
| Regulatory Status | RUO |
| Sensitivity | ＜50pg/mL |
| Standard Curve Range | 50 pg/mL-3200 pg/mL |
| Assay Time | 2 hr |
| Suitable Sample Type | For the quantitative determination of recombinant protein L |
| Sample volume | 50uL |
背景（Background）
Protein 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).
应用说明（Application）
The kit is developed for the detection of natural or structurally conserved recombinant forms of Protein L and a recombinant form of Protein L with very significant structural differences from natural Protein A such as MabSelectTM VL in Bioprocess manufacturing applications. It is used as a tool to aid in optimal purification process development and in routine quality control of in-process streams as well as final product .
It is for research use only.
存储（Storage）
The unopened kit is stable for 12 months from the date of manufacture if stored at 2°C to 8°C.
The opened kit should be stored per components table. The shelf life is 30 days from the date of opening.
组分（Materials Provided）
| ID | Components | Size |
| RES026-C01 | Pre-Coated Anti-Protein L Antibody Microplate | 1plate |
| RES026-C02 | Recombinant Protein L Standard (1μg/mL) | 100μL |
| RES026-C03 | Biotin-Anti-Protein L Antibody | 150uL |
| RES026-C04 | Streptavidin-HRP | 10μg |
| RES026-C05 | 10×Sample Dilution Buffer | 15mL |
| RES026-C06 | Denaturation Buffer | 15mL |
| RES026-C07 | 20×Washing Buffer | 30mL |
| RES026-C08 | Antibody Dilution Buffer | 15mL |
| RES026-C09 | Streptavidin-HRP Dilution Buffer | 15mL |
| RES026-C10 | Substrate Solution | 12mL |
| RES026-C11 | Stop Solution | 8mL |
原理（Assay Principles）
The resDetect™ Protein L ELISA Kit is used to measure the levels of recombinant Protein L by employing a standard sandwich-ELISA format. The micro-plate in the kit has been pre-coated with anti- Protein L polyclonal antibody. Firstly, the standard samples provided in kit and your samples are treated with Denaturation Buffer to dissociation of Protein L and antibody, stand a few minute. Before adding standards and samples, add the Biotin-Anti- Protein L Antibody to the plate to ensure that the standard samples are neutralized by the Biotin-Anti- Protein L Antibody buffer solution and protect the pre-coated antibody on the plate. Then, add the standard samples and your samples to the plate and form Antibody-antigen (Protein L) - biotinylated antibody complex, incubate and wash the wells. Next add Horseradish peroxidase conjugated streptavidin (Streptavidin-HRP) to the plate, incubate and wash the wells to remove any unbound reactants. At last, load the tetramethylbenzidine (TMB) substrate into the wells and monitor a blue color. The reaction is stopped by the addition of a stop solution and the color turns yellow. The intensity of the absorbance can be measured at 450nm and 630nm on a microtiter plate reader. The OD Value reflects the amount of Protein L.
典型数据-Typical Data Please refer to DS document for the assay protocol.
Detection of Recombinant Protein L by sandwich-ELISA Assay. Immobilized Anti- Protein L Antibody can bind Recombinant Protein L . Detection was performed using Biotin-Anti- Protein L Antibody with sensitivity of 50 pg/mL. For each experiment, a standard curve needs to be set for each micro-plate, and the specific OD value may vary depending on different laboratories, testers, or equipments. The following example data is for reference only.
验证（Validation）
批内差异（Intra-Assay Statistics）
Three samples of known concentration were tested ten times on one plate to assess intra-assay precision.
批间差异（Inter-Assay Statistics）
Three samples of known concentration were tested in three separate assays to assess inter-assay precision.
回收率（Recovery）
Add different concentrations of Protein L (0.2ng/mL、1ng/mL、10ng/mL) to different concentrations of Human IgG1 (Bevacizumab) (20mg/mL、10mg/mL、5mg/mL) or Human IgG4 (Toripalimab) (10mg/mL、5mg/mL、2mg/mL), then dilute the antibodies to a reasonable range, then test and calculated the concentration of protein L to give the recovery rate.
干扰效应(Interference effect)
We have conducted interference effect test about frequently-used buffers, they have excellent buffer compatibility. For specific buffers, it is recommended that you verify recovery to determine the minimum dilution ratio.
专属性（Specificity）
Host cell protein (HCP 500 ng/mL) and host cell DNA (HCD 0.5 ng/mL) of HEK293, E.coli or CHO systems were added to human IgG1 (Bevacizumab, 1mg/mL) and human IgG4 (Toripalimab, 1mg/mL), respectively, which were higher than the usual quality standard limit. Then high, medium, and low concentrations of Protein L were added, respectively, and the ratio of Protein L recovery in the Protein L added samples without HCP and HCD was added as the specificity verification index. verification index.
背景（Background）：protein L
Protein 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).
