Rabbit Polyclonal Anti-Collagen IV Antibody from Abcam

Abcam’s anti-collagen IV antibody is designed to bind particularly to native collagen epitopes. It is used for investigating key structural components of glomerular basement membranes (GBM).

  • Product Name - Anti-Collagen IV antibody
  • Description - Rabbit polyclonal to Collagen IV
  • Host Species - Rabbit
  • Tested Applications - Suitable for IHC-FoFr, IHC-FrFl, IHC-P, ICC/IF, WB, IP, IHC-Fr, and ELISA.
  • Species Reactivity - The antibody reacts with various species like human, hamster, mouse, rat, dog, cow, pig, Syrian hamster, Chinese hamster, African green monkey, and zebrafish. It is expected to work with mammals.
  • Immunogen - It is a full-length native protein (purified) that corresponds to Collagen IV, which is derived from bovine and human placenta. The immunogen preserves the protein’s native conformation.
  • Positive Control - Natural Human Collagen IV protein (ab7536) can be utilized as a positive control in WB. It is a human epidermal keratinocytes lysate.

Specificity

The ab6586 antibody is designed to adhere precisely to native collagen epitopes made up of numerous subunit strands. It has negligible cross-reactivity with Type VI, V, III, II, or I collagens. In addition, the non-specific cross-reaction of anti-collagen antibodies with other types of non-collagen extracellular matrix proteins or human serum proteins is insignificant.

General Notes

A minimum of 11 genetically different gene products is collectively called “collagen types” or other proteoglycans and proteins of the extracellular matrix. Collagens in humans are made up of around 20 special protein chains that go through different types of post-translational modifications and are eventually organized into a triple helix. This leads to a major difference between numerous types of collagens.

Greatly conserved throughout evolution, collagens are defined by a continuous “Glycine-X-Y” triplet repeat that forms an essential component of the triple helical structure. Owing to these reasons, it is often highly complicated to produce antibodies that have specificities to collagens. Non-denatured three-dimensional (3D) epitopes govern the development of type-specific antibodies. This preparation leads to native conformation of the protein.

The antibody is ideal for detecting extracellular matrix proteins in both diseased and normal state tissues. Neoplasia is mainly characterized by the disruption of tissue organization. It is possible to distinguish malignant lesions from benign, by analyzing the breakdown of basement membranes and loss of 3D architecture.

It is believed that malignant cells utilize matrix metalloproteases to break down barriers generated by the extracellular matrix, which subsequently enables metastasis. Gelatinases, stomelysins, and collagenases can collectively decompose all the numerous components of the extracellular matrix, such as basement membrane glycoproteins, non-fibrillar collagens, and fibrillar collagens.

Abcam recommends Goat Anti-Rabbit Alexa Fluor® 488 (ab150077) and Goat Anti-Rabbit HRP (ab205718) secondaries.

Applications

Abcam’s Abpromise guarantee covers the usage of ab6586 in the tested applications listed in the table below. Recommended starting dilutions have been mentioned; the end-user should determine the ideal concentrations/dilutions.

Application Abreviews Notes
ELISA 1/5000–1/50000
IHC-Fr ☆☆☆☆☆ 1/50–1/200
IP ☆☆☆☆☆ 1/100
WB ☆☆☆☆☆ 1/1000–1/10000. Should be used under non-reducing conditions. Expected molecular weight is 160 kDa.
However, the product is not suitable for use under denaturing conditions in ELISA, IP, and WB. Hence, Abcam recommends testing it under native conditions.
ICC/IF ☆☆☆☆ Should be used at an assay dependent concentration. PubMed: 19933193.
IF Should be used at an assay dependent concentration. PubMed: 28153846.
IHC-P ☆☆☆☆☆ 1/500. Heat-mediated antigen retrieval should be performed prior to starting with IHC staining protocol.
IHC-FrFl ☆☆☆☆☆ Should be used at an assay dependent concentration.
IHC-FoFr ☆☆☆☆☆ Should be used at an assay dependent concentration.

Target

Function

Here, type IV collagen happens to be the main structural component of glomerular basement membranes (GBM), creating a “chicken-wire” meshwork along with nidogen/entactin, proteoglycans, and laminins.

Arresten, containing the C-terminal NC1 domain, suppresses angiogenesis as well as tumor formation. It was found that the C-terminal half possesses the anti-angiogenic activity. It particularly suppresses the proliferation, migration, and tube formation of endothelial cells. It also prevents the expression of hypoxia-inducible factor 1-alpha and activation of ERK1/2 and p38 MAPK. It is a ligand for alpha-1/beta-1 integrin.

Involvement in Disease

COL4A1 defects lead to brain small vessel disease with hemorrhage (BSVDH) [MIM:607595]. BSVDH account for a larger proportion of intracerebral hemorrhages and underlie 20%–30% of ischemic strokes. Inheritance is autosomal dominant.

COL4A1 defects also lead to hereditary angiopathy with muscle cramps and nephropathy aneurysms (HANAC) [MIM:611773]. Bilateral large cysts and hematuria constitute the clinical renal manifestations. Complex basement membrane defects in the skin and kidney were revealed by histologic analysis. The systemic angiopathy seems to have an impact on large arteries as well as small vessels.

COL4A1 defects lead to porencephaly familial (PCEPH) [MIM:175780] - a term employed for any cerebrospinal fluid-filled cyst or cavitation in the brain. Porencephaly type 1 is often unilateral and arises from focal destructive lesions like birth trauma or fetal vascular occlusion. Schizencephalic porencephaly, or Type 2, is often symmetric and denotes a major arrest or defect in the cerebral ventricle development.