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Antibody > Antibody Labelling > Flashcards

Antibody Labelling Flashcards

1
Q

2 types of Labelling

A

Monoclonal

Polyclonal

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2
Q

How are Antibody Labelling characterised by?

A
  • Non covelent interaction (similar to the lock and key)
  • Detection of Ag or Ab
  • Diagnosis of disease
  • Identify molecular biology and medical intrest
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3
Q

What is the basic concept of immunological assays?

A

Antigen A+Anti- Antigen A= Positive interaction

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4
Q

What celluar antigens can we target?

A 
Cytoplasmic
Nuclear
Cell membrane
Lipids
Proteins
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5
Q

Why is labelled material usually used?

A

used to measure the amount of antigen or antibody present

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6
Q

3 labelling methods?

A
  1. Immunoenzyme methods
  2. Immunofloresence methods
  3. Immuofold methods
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7
Q

description of the immunoenzyme method?

A

Enzyme used as a label

eg, Peroxidase, Glucose Oxidase

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8
Q

description of the immunofloresence method?

A

Flourochrome used as a label

e,g AMCA, Flourscin, TRITC

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9
Q

Immunogold method description?

A

Collodial gold particles are used as the label

USed in microscopy

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10
Q

Direct labelling methods?

A

Label attached directly to the primary antibody. Used in kidney biopses

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11
Q

Indirect labelling methods?

A

Label attached to a secondary antibody:

  • allows greater amplification
  • More economic
  • Longer
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12
Q

5 Types of antibody load?

A
  1. Enzymes
  2. Biotin
  3. Flourophores
  4. Gold
  5. Radioisotope labels
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13
Q

Enzymes acting as an antibody load description?

A

2 typical substances:

  1. 5-brono-4-chloro-3’indolyphosphate ptolyphosphate salt (BCIP)
  2. Nitro blue tetracolium chloride (NBT)
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14
Q

Preparation for enzyme conjugates?

A

Chemical cross linking

Size exclusion chromatography

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15
Q

Biotin description?

A

Covelently linked to Fc domain using aminocapric acid
Free biotin removed by gel filtration
Optimally 20-30 biotin
Amplification step

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16
Q

What is the signal amplification with biotin labelling?

A

Label attached to a secondary antibody:
-high sensitivity
-Longer + more expensive
greater signal amplification

17
Q

Fluorophores description

A

Excitable molecules with distinct excitation/emission wavelengths–> eg, FITC

18
Q

What can a Flow Cytometer tell us about a cell?

A

Its relative size
Its gravity
Relative flouresence intensity

19
Q

Gold description?

A

Immunoglobulin gold complexes
Antibody + collodial gold + PEG (polyethylene Glycol)
Centrifuge conjugates on sucrose density gradient

20
Q

Radioisotope labels advantages?

A

Flexibility
Size
Sensitivity

21
Q

Radioisotope label disadvantages?

A

Toxicity

Shelf life

Antibody (10 decks)

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