Scribd Logo
Upload

Welcome to Scribd!

  • 54 views

    Affinity of LDH Lab

    Uploaded by

    Zoe A Garwood
    This document summarizes a student's lab report on affinity chromatography to purify lactate dehydrogenase from rat liver supernatant. The student aims to purify LDH and determine the amount retained by the column. Tables show absorbance readings for BSA standards, pyruvate volumes, and LDH activity in fractions. Graphs plot BSA concentration vs absorbance and pyruvate volume vs absorbance. The results will help characterize the purified LDH and interaction with the column.

    Copyright:

    © All Rights Reserved
    Download as DOCX, PDF, TXT or read online from Scribd

    Affinity of LDH Lab

    Uploaded by

    Zoe A Garwood
    54 views5 pages
    This document summarizes a student's lab report on affinity chromatography to purify lactate dehydrogenase from rat liver supernatant. The student aims to purify LDH and determine the amount retained by the column. Tables show absorbance readings for BSA standards, pyruvate volumes, and LDH activity in fractions. Graphs plot BSA concentration vs absorbance and pyruvate volume vs absorbance. The results will help characterize the purified LDH and interaction with the column.

    Original Description:

    Original Title

    Affinity of LDH lab

    Copyright

    © © All Rights Reserved

    Available Formats

    DOCX, PDF, TXT or read online from Scribd

    Did you find this document useful?

    Is this content inappropriate?

    This document summarizes a student's lab report on affinity chromatography to purify lactate dehydrogenase from rat liver supernatant. The student aims to purify LDH and determine the amount retained by the column. Tables show absorbance readings for BSA standards, pyruvate volumes, and LDH activity in fractions. Graphs plot BSA concentration vs absorbance and pyruvate volume vs absorbance. The results will help characterize the purified LDH and interaction with the column.

    Copyright:

    © All Rights Reserved
    Download as DOCX, PDF, TXT or read online from Scribd
    Download as docx, pdf, or txt
    54 views5 pages

    Affinity of LDH Lab

    Uploaded by

    Zoe A Garwood
    This document summarizes a student's lab report on affinity chromatography to purify lactate dehydrogenase from rat liver supernatant. The student aims to purify LDH and determine the amount retained by the column. Tables show absorbance readings for BSA standards, pyruvate volumes, and LDH activity in fractions. Graphs plot BSA concentration vs absorbance and pyruvate volume vs absorbance. The results will help characterize the purified LDH and interaction with the column.

    Copyright:

    © All Rights Reserved
    Download as DOCX, PDF, TXT or read online from Scribd
    Download as docx, pdf, or txt
    of 5

    Student’s Name: Zoe Garwood

    Student’s ID#: 620137260

    Lab date: October 27, 2021

    Lab Number: 7

    Lab Title: Affinity Chromatography of Lactate Dehydrogenase

    Aim:
    1. To purify Lactate Dehydrogenase from rat liver supernatant
    2. To determine the amount of interacting proteins that can be retained by the column
    Methodology:
    Method is as seen in the department of Biochemistry Laboratory Manual
    Results:
    Table 1 showing the Absorbance values at 595nm versus the BSA concentration

    BSA 1 2 Avg
    concentration/Mg/mL

    0.00 0.000 0.000 0.000

    0.05 0.039 0.046 0.043

    0.10 0.088 0.075 0.082

    0.15 0.105 0.114 0.110

    0.20 0.155 0.161 0.158

    0.40 0.305 0.293 0.299

    0.60 0.402 0.417 0.410

    0.80 0.514 0.505 0.510

    1.00 0.625 0.610 0.618


    Table 2 showing Volume of Pyruvate in mLs versus the absorbance reading at 440nm

    Pyruvate Absorbance reading at 440nm Avg


    Volume/mLs

    1 2 Avg

    0.0 0 0 0

    0.1 0.038 0.042 0.040

    0.2 0.084 0.082 0.083

    0.3 0.106 0.112 0.109

    0.4 0.183 0..167 0.175

    0.5 0.218 0.203 0.211


    Table 3 showing the absorbance reading at 595nm for the LDH activity for the control, supernatant and
    the fractions using the red and green dye

    Absorbance at 595nm

    Test Sample Red Dye Green Dye

    Control blank blank

    Supernatant 0.299 0.249

    Fraction1 0.106 0.162

    Fraction 2 0.056 0.105

    Fraction 3 0.064 0.160

    Fraction 4 0.045 0.120

    Fraction 5 0.248 0.124

    Fraction 6 0.077 0.125


    Graph 1 showing BSA Concentration versus absorbance
    reading at 595nm
    0.7

    0.6

    0.5

    0.4

    0.3

    0.2

    0.1

    0
    0 0.2 0.4 0.6 0.8 1 1.2

    1 2 Avg

    Graph 2 showing the Pyruvate Volume versus the Absorbance


    reading at 440nm
    0.25

    0.2

    0.15

    0.1

    0.05

    0
    0 0.1 0.2 0.3 0.4 0.5 0.6

    Absorbance @440nm 1 Absorbance @440nm 2 Absorbance @440nm Avg

    You might also like