ELISA Tutorial 5: Preparing ELISA Data in Excel for Analysis with GraphPad Prism

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Here, we demonstrate the first steps of data analysis for some typical ELISA data. The procedure shown is as follows: 1) Compare the arrangement of our samples on the 96-well plate with the absorbance measurements obtained from the plate reader. 2) Subtract out the blank measurements from all the entire plate. 3) Arrange the absorbance measurements of the standards next to their corresponding known concentrations. 4) Place the absorbance measurements of the samples below the absorbance measurements of the standards.
In the next video of this series, we will demonstrate how to interpolate the concentration of our samples using software called GraphPad Prism. Click here for a free 30-day trial of GraphPad Prism - protocol-place.com/Prism
This video and other protocols can be found at our website, the "Protocol Place" - protocol-place.com/
A full ELISA protocol can be found at protocol-place.com/assays/sand...
This video is a part of our ELISA Tutorial Series. Here are all of the videos in this series:
ELISA Tutorial 1: Understand How an ELISA Works - • ELISA Tutorial 1: How ...
ELISA Tutorial 2: Coating and Blocking the ELISA Plate - • ELISA Tutorial 2: Coat...
ELISA Tutorial 3: Preparing and Adding Samples to the ELISA Plate - • ELISA Tutorial 3: Prep...
ELISA Tutorial 4: Finishing the Assay (Sandwich ELISA) - • ELISA Tutorial 4: Fini...
ELISA Tutorial 5: Preparing ELISA Data in Excel for Analysis with GraphPad Prism - • ELISA Tutorial 5: Prep...
ELISA Tutorial 6: How to Analyze ELISA Data with GraphPad Prism - • ELISA Tutorial 6: How ...
Competitive ELISA Tutorial 1: How a Competitive ELISA Works - • Competitive ELISA Tuto...
Competitive ELISA Tutorial 2: How to Use Calbiotech's Competitive ELISA Kits - • Competitive ELISA Tuto...
Competitive ELISA Tutorial 3: Analyzing Typical ELISA Data in Excel - • Competitive ELISA Tuto...
We hope you enjoy watching and benefit from our tutorials. If so, please take a minute to "like," or better yet, share them with others!
Thanks for watching!
Youssef Farhat
Protocol Place: protocol-place.com
**Check out some of our other tutorials via the links below**
Competitive ELISA Tutorials - - - • Competitive ELISA Tuto...
ELISA Tutorials - - - • ELISA Tutorial 1: How ...
Gelatin Zymography Tutorials - - - • Gelatin Zymography Tut...

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Комментарии : 32

@xma3559 7 лет назад

Very comprehensive. Thanks to your video. I was quite stuck with my assignment!

@sirinamuntaka6146 9 лет назад

very wonderful and simplified tutorial. I loved it

@sirinamuntaka6146 9 лет назад

ery wonder and simplified tutorial. I loved it

@jesuspenaarias9366 7 лет назад

nanananah esto es un plagio barato que gracias ni que leches

@brianleon4575 4 года назад

thanks a lot! this saved me

@felixleaorodriguezfierros1859 6 лет назад

Hi, thank you for the video It was very useful. One question, why are you dividing 500/3??? Thank you for the information

@issame945 2 года назад

vielen Dank. It´s realy very helpfull and easy to understand

@worldofinformation815 2 года назад

Thanks for your video✌✔🌹

@souadalshubrami9099 5 лет назад

thank you for very helpful tutorial , i want to ask how can calculate Absorbance data from Concentration value ?? Back up Equation

@bibianapardo123 Год назад

Thanks a lot for your help, is so importan to my thesis

@qinzhou1927 2 года назад

Thank you so much Very helpful. What was the BLANK? was it your negative?

@alvinmagana3356 Год назад

Great job

@spkspk1416 3 года назад

Thank you!

@cdscodi1 3 года назад

What is gray zone calculations that plus - minus 10% of CO value which is mentioned in literature of Elisa

@JBakedDeans 4 года назад

Thanks McLovin

@sharon_ng 7 лет назад

Why do you not make the -ve values into 0 since it can't have less than 0 antibody?

@ishtiaqahmed3785 Год назад

I have IDEXX ELISA kit for paratuberculosis. Formula is given there on brochure. OD values have to be incorporated in formula. But end results are confusing. I need help please.

@apekshaannigeri8532 4 года назад

it is very usefull snd simplfied. thanks a lot. but then i have a query. why do you have to subtract the blank value from all the sample ODs?

@wc389 4 года назад

In a perfect world, the blank well would read 0.0000, however this never happens, depending on the material of the plate, the temperature of the reader, the type of reader, each well will give off an absorbance value. This suggests that the blank well has some of your target analyte in it. However if you have completed the experiment correctly, your blank well does not have any analyte in it. So we must remove the value of the blank from each of the wells, to correct for atmospherics...

@esmatshaker6073 8 лет назад

thank u so much , i m sorry i dont understood the part of concentration about 500 what is this number and how u r get it

@jontrip247 8 лет назад

+esmat shaker Nanodrop is one method to obtain concentrations. There are others

@ytlau2247 3 месяца назад

Hi is this applicable to direct ELISA too?

@pksaini8107 2 месяца назад

.TNF-alpha= 450 value - 0.132 Concentration - 11.792 Inme se result konsa hai.. please tell me

@abdulwahabkhashab2933 8 лет назад

Very informative indeed, but i wonder why did you subtract your blank using your average blank?????

@Protocolplace-about 8 лет назад

Blank measurements have some variation to them, so subtracting an average is a more accurate way to account for the assay's background signal.

@abdulwahabkhashab2933 8 лет назад

Even with the blank itself!!!!! this is new to me.

@Protocolplace-about 8 лет назад

+abdulwahab KHASHAB Yes, if you would like to use the blank measurements on the standard curve that you use to interpret your data, then you should most certainly subtract the blank average from the blank values themselves.

@abdulwahabkhashab2933 8 лет назад

I'll try it out and compare both curves. Thanks allot

@sharon_ng 7 лет назад

Why are there -ve values?

@MrIraqyforlife 2 года назад

thx

@yjharvey7824 2 года назад

I think you made a mistake calculating the background. You need to type in =average (b9:m9) but yours is =average (b9,m9) so you actually only selected two cells which made the background average very low.