2 - PROTEIN (Lab Methods Only)

Protein electrophoresis (MOST → LEAST anodal)

Albumin → Alpha 1 → Alpha 2 → Beta → Gamma

INCREASED: alpha 1 and alpha 2

Acute Inflammation

INCREASED: alpha 1 & alpha 2, & gamma

Chronic Infection

PROTEIN ELECTROPHORESIS:

• Polyclonal with BETA-GAMMA BINDING

Cirrhosis

PROTEIN ELECTROPHORESIS:

• Sharp ↑ in immunoglobulin “M Spike”

Monoclonal gammopathy

PROTEIN ELECTROPHORESIS:

• low albumin

• high alpha 2

Nephrotic Syndrome

REFERENCE METHOD for protein determination

• KJELDAHL method

  • H2SO4 + CuSO4

Biuret → most common method

PROTEIN ELECTROPHORESIS:

• SALT PRECIPITATION METHOD:

  • what are precipitated in high salt concentration?

  • where is albumin found?

  • albumin is quantified by?

  • reagents used?

PROTEIN ELECTROPHORESIS:

• Globulins = precipitated

• Albumin = in supernatant (quantified by Biuret Rxn)

  • Sodium Sulfate

  • Sodium Sulfite

  • Ammonium Sulfate

  • Methanol

PROTEIN DYE BINDING:

• Non-specific dye for albumin

• Methyl Orange → Non-specific
  
  

• HABA → Specific but subject to interferences

• Bromocresol Green (BSG) → most common (not affected)

• Bromocresol Purple (BSP) → Specific & Precise

PROTEIN DYE BINDING:

• Specific for albumin but subject to interferences (SALICYLATES)

• Methyl Orange → Non-specific

• HABA → Specific but subject to interferences

• Bromocresol Green (BSG) → most common (not affected)

• Bromocresol Purple (BSP) → Specific & Precise

PROTEIN DYE BINDING:

• Most common method used

• Not interfered by SALICYLATES

• Methyl Orange → Non-specific

• HABA → Specific but subject to interferences

• Bromocresol Green (BSG) → most common (not affected)

• Bromocresol Purple (BSP) → Specific & Precise

PROTEIN DYE BINDING:

• SPECIFIC and PRECISE

• Methyl Orange → Non-specific

• HABA → Specific but subject to interferences

• Bromocresol Green (BSG) → most common (not affected)

• Bromocresol Purple (BSP) → Specific & Precise