Question

I am stuck on how to find concentration for standards and unknowns. Any kind of help would be appreciated.

The corrected absorbance is = Blank - absorbance

1. Prepare the BCA working reagent. Use a graduated cylinder and add 30 ml of the BCA reagent into a 50 ml Erlenmeyer flask. Use the appropriate micropipette to add 600 uL of the CuSO4 solution into the flask and mix by swirling the flask. Record the exact mass of albumin and the volume of the volumetric flask used to prepare the -1.xx mg/mL protein stock solution. This information is needed to calculate the exact concentration of the stock solution. Prepare a -0.10xx mg/mL protein working standard solution from the stock solution. Transfer 1.000 mL of the stock solution to a 10.00 mL volumetric flask using the appropriate micropipette. Dilute to volume with DI water. Pour the working standard solution into a clean, dry test tube. If the test tube is wet, first rinse with a portion of the working standard before pouring the remaining solution into the test tube. Place the tube in the test tube holder. Prepare your blank and standard samples according to the table below. NOTE: All samples (blank, standards, and unknown) are prepared in test tubes. 6. Select one unknown sample that you will analyze in triplicate. Record the unknown number. Transfer 0.500 mL of the unknown into a clean dry test tube using the appropriate micropipette. Prepare the unknown sample solution in triplicate. Cut out and paste this table into your lab notebook Volume of Volume Working Protein Volume BCA Sample Total Unknown Std Solution Volume DI Working Volume Water (mL) Solution (mL) Reagent (mL) (mL) (mL) Blank 3.000 1.000 4.000 Standard 1 0.100 0 3.000 0.900 4.000 Standard 2 0.200 0 3.000 0.800 4.000 Standard 3 0.300 0 3.000 0.700 4.000 Standard 4 | 0.400 0 3.000 0.600 4.000 Standard 5 0.500 0 3.000 0.500 4.000 Unknown 0.100 3.000 0.900 4.000 Unknown 0.100 3.000 0.900 4.000 Unknown 0.100 3.000 0.900 4.000 Cover the top of the tubes with Parafilm. Mix each sample for -10-15 seconds using a vortex mixer. Place the tubes in the constant temperature water bath at 60°C. While waiting for the samples to complete the 30 min. reaction time, turn on the LabQuest instrument. Complete operating instructions I and II on the next page. After 30 minutes, remove the samples from the water bath and cool to room temperature. 0 0

Answer 1

০০a3 6 Cstecs 1-0034 mty Soome Toras for Standard volume ntcnwn 4-o00 CSum o used Soluvans of all volume Csrondad arant of Shoxkadded Tor volume 1-0034M X oloo ml 2 O-0252 Mg/m t-004 2) Estandard 0504 m9/m Cskndard 3) 1-0031 0300ml 2 9-000 m mgIm 1ిం KO.40oml Csrandaud y mg ImL O-101 x0-406 ml +-0024 mg Csrandcrd msLmL O-126 Cae Sandaud Cons enhaen 2 ab scabance (Y-vales comeched 0131-0-134 -333 - ০-1১৭ 6-159 O-O So4 mg(mt Oh24-0-(31 245 O0756 mg/m O-40) l01 mym ৩१०৪ - 0-139 o-S29 o-126 9/me ৪५० -0121 O-661

taking cbsabnce dvaw gyaph When he cansenhahen Cnd Valye On of voten Valces ecreatn gek we +0-006 049 5-139014 x 2. S-18 x +o-006 of dilaed unknowA 1 Solon And Concentraton absovbunce 0-139 zo-238 Covected u get x ১১bshe -124014xto-006049 2 0-0101g O.olo22 ng i. oalo Solunen ongir of prcten Concencentaha mass hooles Pre sent luted unIcnown mass lginal added ఎ 22 XY-00 ml MC O.4088 mgLmL CPvdenn n ngira unknown O41 mg(m 2