Rat Laminin ELISA Kit

Catalog No.: BEK1135

Size: 96T

Range: 156 pg/ml-10,000 pg/ml

Sensitivity < 10 pg/ml

Storage and Expiration: Store at 2-8℃ for 6 months, or at -20℃ for 12 months.

Application: For quantitative detection of Laminin in rat serum, plasma, body fluids, tissue lysates or cell culture supernatants.

Introduction

Laminins are major proteins in the basal lamina, a protein network foundation for most cells and organs. Fifteen laminin trimers have been identified. The laminins are combinations of different alpha-, beta-, and gamma-chains. The laminins are an important and biologically active part of the basal lamina, influencing cell differentiation, migration, adhesion as well as phenotype and survival. Recently, several publications have demonstrated that laminins can be used to culture cells, such as pluripotent stem cells, that are difficult to culture on other substrates. Rodin et al. showed that recombinant laminin 511 can be used to create a totally xeno-free and defined cell culture environment to culture human pluripotent ES cells and human iPS cells.

Principle of the Assay  

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-Laminin polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-Laminin polyclonal antibody was used as detection antibodies. The standards, test samples and biotin conjugated detection antibody were added to the wells subsequently, and wash with wash buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with wash buffer. TMB substrates were used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the Laminin amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of Laminin can be calculated.

Kit components

1. One 96-well plate pre-coated with anti-Rat Laminin antibody
2. Lyophilized rat Laminin standards: 2 tubes (10 ng / tube)
3. Sample / Standard diluent buffer: 30ml
4. Biotin conjugated anti-rat Laminin antibody (Concentrated): 130μl. Dilution: 1:100
5. Antibody diluent buffer: 12ml
6. Avidin-Biotin-Peroxidase Complex (ABC) (Concentrated): 130μl. Dilution: 1:100
7. ABC diluent buffer: 12ml
8. TMB substrate: 10ml
9. Stop solution: 10ml
10. Wash buffer(25X): 30ml

Note: Reconstitute standards and test samples with Kit Component 3.

Material Required But Not Provided

1. 37℃ incubator
2. Microplate reader (wavelength: 450nm)
3. Precise pipette and disposable pipette tips
4. Automated plate washer
5. ELISA shaker
6. 1.5ml of Eppendorf tubes
7. Plate cover
8. Absorbent filter papers
9. Plastic or glass container with volume of above 1L