For staining human renal and skin biopsy sections easily and rapidly!

FITC-Anti Collagen IV a5(IV) Chain, Human (Mono)+Texas Red-Anti Collagen IV a2(IV) Chain, Human (Mono)

Background

This product can be used for staining human renal and skin biopsy sections easily and rapidly as two monoclonal antibodies are conjugated with two different fluorochromes. FITC-conjugated-anti a5(IV) demonstrates the presence or absence of the a5(IV) chain, and Texas Red –anti a2(IV) reveals the renal basement membrane structure.

Alport syndrome, an inherited disease, shows the absence or reduction of the a5(IV) chain in the glomerular, tubular and Bowman’s capsular basement membranes. Normal human kidney have a1(IV) to a6(IV) chains in the renal basement membranes.

Double exposure of normal human kidney section stained with the product. FITC fluorescence (a5 chain) is observed in the GBM, part of the TBM and Bowman’s capsular BM. BMs were two fluorochromes are present look orange to yellow.

Double exposure of Alport human kidney section stained with the product. Because no FITC fluorescence (a5 chain) is observed in the kidney of the patient with X-linked Alport'fs syndrome, only Texas Red fluorescence (a2 chain) is confirmed.

Application Procedures

Protocol

CFT-45325 is prepared to stain human cryostat sections of biopsy specimens from kidney and skin by direct immunofluorescence.

Staining Examples [PDF]

How to stain human cryostat sections
Wash cryostat sections with phosphate buffered saline (PBS), wipe PBS around them with a paper wiper, and incubate them with 20-50 micro liter of the staining solution for 30-60 min at room temperature. After washing them with PBS, mount them and observe them with a fluorescence microscope

Cautions

• Do not dilute the staining solution.
• Do not use stale cryostat sections.
• Do not fix sections with any fixatives.
• In case that a section is not stained or very poorly stained with FITC-anti-a5(IV) but the background staining of Texas Red-anti-a2(IV) is normally stained
  1) The disease is diagnosed as Alport syndrome. Or,
  2) There is still a possibility that a yet-unknown amino acid substitution is present in the region of the epitope sequence.
• For accurate diagnosis, it is important to consider clinical symptoms, and to make additional staining with other monoclonal antibodies against type IV collagen.

 

Product Name	Cat#	Quantity
Fluorochrome-conjugated Anti Collagen IV cocktail for Alport's syndrome	SGE-CFT45325	1ML

Collagen Detection Antibodies and Kits Flyer [PDF]

 

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