PRINCIPLE

Enzyme-Linked Immunosorbent Assay (ELISA) is a plate-based assay technique, which is also named enzyme immunoassay (EIA). The technique was designed for detecting and quantifying substances such as proteins, antibodies and hormones. In an ELISA, antigen from the sample is attached to a solid surface and then complexed with an antibody. This antibody is linked to an enzyme which can catalyze substrates and produces detectable signal, most commonly a color change.

Material

Reagents Instruments and Consumables
Coating Buffer Coated Antigen Microplate Reader (CMax Plus)
Blocking Buffer Detect Antibody Eppendoff
Washing Buffer HRP-conjugated Secondary Antibody Constant Temperature Incubator
TMB Solution
Stopping Buffer

METHODS

  1. Coat microtiter plate wells with 100 µl of the antigen at a concentration (e.g. 2 µg/mL) in coating buffer. Incubate overnight at 4°C. Wash the plate 3 times with washing buffer.
  2. Add 200 µl of blocking buffer to each well. Incubate for 2 hours at 37°C. Wash 4 times.
  3. Add 100 µl of diluted Antibody to the relevant wells. Incubate for 1 hour at 37°C. Wash 4 times.
  4. Add 100 µl of HRP-conjugated secondary antibody to each well. Incubate for 1 hour at 37°C. Wash 4 times.
  5. Add 100 µl of TMB solution to each well. Incubate at 37°C for 10-15 minutes.
  6. Add 50 µl of stop buffer. Gently tap plate to ensure thorough mixing.
  7. Read the OD Value (450 nm).
  8. Plate configuration.

HOT PRODUCTS

PDCD1 HIV1 gp120 CD33 IAV HA PRNP TNF MS4A1 CD274 APP EGFR CD40 ERBB3 TFPI ERBB2 SARS-CoV S PCSK9 MET BSG HAVCR2 IL6 CD3 LAG3 ROR1 TNFRSF4 IL33 CD19 CD38 JAG1 IL17A IFNG CD47 HCV E2 Protein IL1A / IL1B HBsAg IGF1R CD4 AXL ANGPT2 IL23A NGF MUC1 TNFRSF18 TNFRSF9 MSTN IL1B HRSV F Protein KDR SORT1 C5 VEGF CXCR4 TNFRSF17 Tau TFRC CTLA4 IL13 IAV H5N1 HA TSLP MSLN SOST
For research use only, not directly for clinical use.
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