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For staining human renal and skin biopsy sections easily and rapidly! FITC-Anti Collagen IV α5(IV) Chain, Human (Mono)+Texas Red-Anti Collagen IV α2(IV) Chain, Human (Mono)

Alport's Syndrome Monoclonal Staining Cocktail

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.

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-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.

 Collagen Detection Antibodies and Kits Flyer [PDF]
  1. Cell Matrix Protein Antibody (Anti Collagen)
  2. Cell Matrix Protein Antibody (Others)
  3. Bone Marker Protein Antibody
  4. Visual Protein Antibody
  5. Enzyme Antibody
  6. Homeobox Protein Antibody
  7. anti human collagen IV a5 antibody
  8. anti collagen 7 monoclonal antibody
  9. Alport's Syndrome Monoclonal Staining Cocktail

Product List

Product Name Cat# Quantity Price

Anti Collagen 4 cocktail for Alport's Syndrome

SGE-CFT-45325 1ML

¥ 55,000
$ 734
€ 550

Citation
  • Negative Staining for COL4A5 Correlates With Worse Prognosis and More Severe Ultrastructural Alterations in Males With Alport Syndrome
    Samar M. Said, Mary E.Fidler, Anthony M. Valeri, Brooke McCann, Wade Fiedler, Lynn D.Cornell, Mariam Priya Alexander, Ahmed M. Alkhunaizi, Anne Sullivan, Carl H. Cramer, Marie C. Hogan, Samih H. Nasr
    Kidney Int Rep. 2017 Jan;2(1):44-52. PMID:
  • Functional assessment of a novel COL4A5 splice region variant and immunostaining of plucked hair follicles as an alternative method of diagnosis in X-linked Alport syndrome.
    Malone AF, Funk SD, Alhamad T, Miner JH.
    Pediatr Nephrol. 2017 Jun;32(6):997-1003. PMID: 28013382
  • Alport syndrome and pregnancy: Good obstetric and nephrological outcomes in a pregnant woman with homozygous autosomal recessive Alport syndrome.
    Nishizawa Y, Takei T, Miyaoka T, Kamei D, Mochizuki T, Nitta K.
    J Obstet Gynaecol Res. 2016 Mar;42(3):331-5. PMID: 26628290
  • X-linked Alport syndrome caused by splicing mutations in COL4A5.
    Nozu K, Vorechovsky I, Kaito H, Fu XJ, Nakanishi K, Hashimura Y, Hashimoto F, Kamei K, Ito S, Kaku Y, Imasawa T, Ushijima K, Shimizu J, Makita Y, Konomoto T, Yoshikawa N, Iijima K.
    Clin J Am Soc Nephrol. 2014 Nov 7;9(11):1958-64. PMID: 25183659
  • Pregnancy complicated with Alport syndrome: a good obstetric outcome and failure to diagnose an infant born to a mother with Alport syndrome by umbilical cord immunofluorescence staining.
    Matsubara S, Ueda Y, Takahashi H, Nagai T, Kuwata T, Muto S, Yamaguchi T, Takizawa T, Suzuki M.
    J Obstet Gynaecol Res. 2009 Dec;35(6):1109-14. PMID: 20144175
  • Collagen alpha5 and alpha2(IV) chain coexpression: analysis of skin biopsies of Alport patients.
    Patey-Mariaud de Serre N, Garfa M, Bessieres B, Noel LH, Knebelmann B.
    Kidney Int. 2007 Aug;72(4):512-6. PMID: 17554254
  • Autosomal dominant Alport's syndrome: study of a large Tunisian family.
    Kharrat M, Makni S, Makni K, Kammoun K, Charfeddine K, Azaeiz H, Jarraya F, Ben Hmida M, Gubler MC, Ayadi H, Hachicha J.
    Saudi J Kidney Dis Transpl. 2006 Sep;17(3):320-5. PMID: 16970251
  • In vivo expression of podocyte slit diaphragm-associated proteins in nephrotic patients with NPHS2 mutation.
    Zhang SY, Marlier A, Gribouval O, Gilbert T, Heidet L, Antignac C, Gubler MC.
    Kidney Int. 2004 Sep;66(3):945-54. PMID: 15327385
  • In vivo expression of putative LMX1B targets in nail-patella syndrome kidneys.
    Heidet L, Bongers EM, Sich M, Zhang SY, Loirat C, Meyrier A, Broyer M, Landthaler G, Faller B, Sado Y, Knoers NV, Gubler MC.
    Am J Pathol. 2003 Jul;163(1):145-55. PMID: 12819019

References
  • The NC1 domain of collagen IV encodes a novel network composed of the alpha 1, alpha 2, alpha 5, and alpha 6 chains in smooth muscle basement membranes.
    Borza DB, Bondar O, Ninomiya Y, Sado Y, Naito I, Todd P, Hudson BG.
    J Biol Chem. 2001 Jul 27;276(30):28532-40. PMID: 11375996
  • Epitope-defined monoclonal antibodies against type-IV collagen for diagnosis of Alport's syndrome.
    Kagawa M, Kishiro Y, Naito I, Nemoto T, Nakanishi H, Ninomiya Y, Sado Y.
    Nephrol Dial Transplant. 1997 Jun;12(6):1238-41. PMID: 9198058
  • Relationship between COL4A5 gene mutation and distribution of type IV collagen in male X-linked Alport syndrome. Japanese Alport Network.
    Naito I, Kawai S, Nomura S, Sado Y, Osawa G.
    Kidney Int. 1996 Jul;50(1):304-11. PMID: 8807602
  • Differential expression of two basement membrane collagen genes, COL4A6 and COL4A5, demonstrated by immunofluorescence staining using peptide-specific monoclonal antibodies.
    Ninomiya Y, Kagawa M, Iyama K, Naito I, Kishiro Y, Seyer JM, Sugimoto M, Oohashi T, Sado Y.
    J Cell Biol. 1995 Sep;130(5):1219-29. PMID: 7657706
  • Type IV collagen alpha 5 chain. Normal distribution and abnormalities in X-linked Alport syndrome revealed by monoclonal antibody.
    Yoshioka K, Hino S, Takemura T, Maki S, Wieslander J, Takekoshi Y, Makino H, Kagawa M, Sado Y, Kashtan CE.
    Am J Pathol. 1994 May;144(5):986-96. PMID: 8178947

To be used for research only. DO NOT use for human gene therapy or clinical diagnosis.