Biochem Lab Midterm

146 on a p20

14.6 microliters

072 on a p200

72 microliters

053 on a p1000

530 microliters

P(20) tip color and range

clear 2-20

P(200) tip color and range

yellow 20-200

P(1000) tip color and range

Blue 100-1000

precise

all points close together

accurate

all points close to the theoretical data

R²=1

percise and accurate

R²=0

Not precise or accurate

what is part A?

Plunger

What is part B?

Friction Ring

What is part C?

Micrometer

What is part D?

Digital Volume Indicator

What is part E?

Shaft

What is part F?

Tip Ejector Button

What is part G?

body

What is part H?

Tip Ejector

What is part I?

Pipette Tip

Affinity Chromatography

Beads are linked to a biomolecule that interacts with the protein

Size exclusion chromatography

Beads with small pores separate based on size. large biomolecules elute first then small

Ion-Exchange Chromatography

positive charged beads interact with negative charged protiens.

Which one elutes first?

hemoglobin 65000 Daltons

myoglobin 17000 Daltons

myosin 180000 Daltons

myosin 180000 Daltons

Which one elutes last?

hemoglobin 65000 Daltons

myoglobin 17000 Daltons

myosin 180000 Daltons

myoglobin 17000 Daltons

what gives hemoglobin/ myoglobin its color?

iron containing heme

what does high altitude do to red blood cells

more are produced

gene therapy

provide working copy of the gene

vitamin B12

breakdown of fats

buffer

liquid used in mobile phase

myoglobin

transports oxygen to muscles

hemoglobin

transports oxygen to tissue

immune system Vs. disease

physically blocks pathogens/ makes molecules and cells that recognize pathogens

immune response

expose the immune system to a weak/ inactive pathogen

how do diseases spread?

vector transfer/ exchange of body fluids/ ingestion of contaminated food or water/ inhalation

diseases that attack the immune system

lupus/ HIV

why would immune systems not work?

genetic disorder/ infection with a pathogen/ immunosuppressive meds/ overreaction to an antigen/ autoimmune disorder

immunosuppressants

immune system wont attack transplant tissue

rapid detection of disease exposure

who gets treated/ minimizing exposure

ELISA

enzyme linked immuno sorbent assay

enzymes in immunoassay

convert substrate into a colored compound

inate immunity

born with immunological defenses

passive immunity

acquisition of antibodies from external source

aquired immunity

specific response to specific foreign substance. born with it but must be activated

humoral immunity

production of antibodies in the blood stream/ lymph that bind to foreign antigens

cell mediated

production of T cells that bind/ destroy infections

antigen

foreign to the body

epitope

single shape on an antigen recognized by antibodies

antibody

first line of defense against invading microorganisms

IgG

most abundant in internal body fluids

IgM

serum responsable for primary immune response

IgA

only antibody passed from mom to baby in external secretion

IgD

regulating immune response

IgD

regulating immune response

IgE

allergic reactions

Macrophages

removes foreign cells from blood/ processing antigens

infection transmission

person to person/ animal to person (Zoonosis)/ environment to person

hypersensitive reaction

immune system overreacts to an antigen

Immunodeficiency

unable to mount effective immune response

primary immunodefficincy

genetic basis

secondary immunodefficency

external cause

anaphylactic 

allergies

cytotoxic

transfusion reactions/ Rh incompatibilty

immune complex

inhaling mold spores

Delayed hyper sensitivity

contact sensitivity

autoimmune disease

immune system mistakes and attacks own body

immunofluorescent assay (IFA)

detected with fluorescently labeled antibodies

agglutination

visible precipitates appear when antibodies and specific antigens come in contact

immunochromatography test

dip-stick immunoassays

microplate test

ELISA detects microbial antigensM

molecular methods

detects microbial RNA/ DNA

microscopy

visual identification with staining/ physical characteristics

live attenuated vaccines

weakened (attenuated) microbes

killed/ inactivated vaccines

microbes killed by heat or chemicals

subunit vaccines

made of pieces of microbes

DNA vaccines

DNA that codes for microbial antigens

mRNA vaccines

mRNA molecules encode the antigen

Antibody Vaccines

construct human monodonal antibodies using recombinant DNA tech

post exposure vaccines

vaccine regimen begun promptly after exposure

polyclonal antibodies

immunize/ fuse cells to create hybridoma/ culture cells in liquid growth medium/ growth medium

Genetically engineering antibodies

antibodies that recognize the antigen attached with a drug

hybridoma immortalization

combining human and mouse genes to camoflauge a mouse monoclonal antibody cheaper and quicker

phage display

phage that binds to a specific antigen and can be used like an antibody

phage library

libraries of billions of potentially useful antibodies created by inserting shuffled genes from billions of humans into the bacteriophage lambda so that the lambda phages display the binding sites from human antibodies on their surface

inderect detection

uses polyclonal secondary antibodies to bind to primary antibodies. only one type of enzyme-linked secondary body is needed

immunostaining

localizes antigens in organelles, cells, tissues, or whole organisms, and can be used to distinguish one cell type from another

immunoblotting/ western blotting

tells protien’s size and relative abundance in a given sample

dot blotting

sample is sotted onto a membrane directly rather than being blotted from a ggel

what are four diffrent ways light can interact with biomolecules?

light scattering/ reflection/ absorption/ transmission

in the bradford assay, the protien-dye complex at 545 mm makes the protien dye compllex blue while the dye alone (without proteins) absorbs blue light at 470 nm making the dye…

reddish brown

what has an unprotonated peak absorbance of 595 nm?

coomassie G-250 dye

what are some colorimetric methods?

biuret/ bradford/ lowry

what is the first step of bradford assay?

preparation of a dillution series of known protein standards and preparation of unknowns

what is the second step of bradford assay?

addition of bradford dye and incubation for >5 min

what is step 3 of the bradford assay?

binding of dye to protein, resulting in a color change and reading of the asorption at A595 in a spec

what is step 4 of the bradford assay?

compilation of the data into a standard curve and unknown protein concentration detemination

what is the primary interaction that occurs between Coomassie G-250 dye and proteins in the Bradford assay?

electrostatic interactions between arginine and sulfate groups

why is it important to use a fresh tip for each standard and sample when adding them to the cuvettes in the Bradford assay?

to avoid cross contamination and ensure accurate readings

the closer the correlation coefficient (R²) is to ___ the better the fit of the standard curve

1

biophotonics

technology that focuses on the interaction of bio materials with light and other forms of radiant energy