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For staining human renal and skin biopsy sections easily and rapidly! FITC-Anti Collagen IV ƒ¿5(IV) Chain, Human (Mono)+Texas Red-Anti Collagen IV ƒ¿2(IV) Chain, Human (Mono)

Fluorochrome-conjugated MoAbs for Alport syndrome


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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 α5(IV) demonstrates the presence or absence of the α5(IV) chain, and Texas Red –anti α2(IV) reveals the renal basement membrane structure.


Alport syndrome, an inherited disease, shows the absence or reduction of the α5(IV) chain in the glomerular, tubular and Bowman’s capsular basement membranes. Normal human kidney have α1(IV) to α6(IV) chains in the renal basement membranes.


Double exposure of normal human kidney section stained with the product.
FITC fluorescence (α5 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 (α5 chain) is observed in the kidney of the patient with X-linked Alport'fs syndrome, only Texas Red fluorescence (α2 chain) is confirmed.


 Collagen Detection Antibodies and Kits Flyer [PDF]

Application Procedures

Protocol [PDF]

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-α5(IV) but the background staining of Texas Red-anti-α2(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.
References

> Kagawa M, et al., (1997) Epitope-defined monoclonal antibodies against type-IV collagen for diagnosis of Alport syndrome. Nephrol. Dial. Transplant. 12: 1238-1241.
> Yoshioka K et al. (1994) Type IV collagen ƒ¿5 chain: Normal distribution and abnormalities in X-linked Alport syndrome revealed by monoclonal antibody. Am. J. Pathol. 144: 986-996.
> Ninomiya Y et al. (1995) Differential expression of two basement membrane collagen genes, COL4A6 and COL4A5, demonstrated by immunofluorescence staining using peptide-specific monoclonal antibodies. J. Cell Biol. 130: 1219-1229.
> Naito I et al., Japanese Alport Network (1996) Relationship between COL4A5 gene mutation and distribution of type IV collagen in male X-linked Alport syndrome. Kidney Int.50: 304-311.

Product List

Product Name Cat# Quantity Price

Anti Collagen 4, Human (Rat) Fluorescein Isothiocyanate/Texas RedTM DataSheet

SGE-CFT-45325

1 ML ¥55,000
$734

Related Products

This product can be used for immunostaining of cryostat sections and for western blotting
Anti Human Collagen IV Alpha5 Chain Monoclonal antibody


Human cryostat section stained with monoclonal antibody H52.
Glomerular basement membrane (BM), part of tubular BM, and Bowman’s capsular BM are stained.

Product Name Cat# Quantity Price

Anti Collagen 4, Human (Rat)  DataSheet

SGE-C-452

500 UL ¥28,000
$374

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