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I need detailed more detailed GENERAL ELISA PROTOCOL including Plate Preparation ,Assay Procedure and TECHNICAL HINTS AND LIMITATIONS for Kallistatin protein (Human) (ab116855) ...online Data sheet is not enough..
Asked on Nov 24 2011
Thank you for contacting us. Here is a protocol that can be applied with this protein (ab116855) to perform an ELISA: 1. Coat antigen (200ng/well) onto the wells in a 96 well mictrotiter plate 2. Block unbound sites with 5% skim milk in PBST 3. Apply primary antibody (diluted in 5% skim milk). 4. Incubate the plate at room temperature for two hours. 5. Wash 4 times with PBST. 6. Apply secondary antibody, and incubate the plate at room temperature for one hour. 7. Wash 4 times with PBST. 8. Apply relevant chromogen and incubate at room temperature for 20 minutes. Read the plate in ELISA reader at 450 nm. A more general protocol for performing ELISAs can be found here: https://www.abcam.com/ps/pdf/protocols/Indirect%20ELISA%20protocol.pdf With a useful troubleshooting guide here: https://www.abcam.com/ps/pdf/protocols/ELISA_troubleshooting.pdf I hope this information has been helpful. If you require any further information please do not hesitate to contact us again.
Abcam Scientific Support
Answered on Nov 25 2011