Useful monoclonal antibodies for the research of cancer pathology
Laminins are major extracellular matrix proteins containing over 15 members. Laminins are large hetero-trimers consisting of 5 alpha chains, 3 beta chains, and 3 gamma chains. Due to its structural varieties, it has been
challenging to detect each laminin isoform by conventional antibodies. Our supplier Funakoshi commercializes 4 unique antibodies, originally developed by Dr. Kaoru Miyazaki, Professor Emeritus of Yokohama City University in Japan. Our antibodies can separately recognize specific isoforms or structures of laminin.
Fig.1 Structure of Laminin 3A32, 3B32 and γ2pf fragment with epitopes of 3 clones (F7, BG5 andP2H).
■ BG5/F7 : For laminin alpha-3 chains
- Laminin alpha 3 chain, a major component of Laminin-332, has two splicing variants, alpha 3A and alpha 3B
- Clone BG5 is specific antibody for alpha 3A
- Clone F7 is the world first commercially available antibody for alpha 3B
■ P2H : For laminin gamma-2 chain
- Laminin gamma2, an only component of Laminin-332, is a marker gene of invasive cancer tissue
- Laminin gamma-2 chain is activated via proteolytical cleavage at N-terminal
- N-terminal proteolytic fragment of gamma-2, called γ2pf, is highly accumulated into cytosol under malignant cancer
- Clone P2H is the world first antibody for γ2pf
■ 12D : For laminin-511
- There are two similar isoforms, laminin-511 and laminin-521 and it is difficult for conventional antibodies such as anti-alpha 5 chain to separate them.
- Clone 12D specifically react with trimeric structure of laminin-511, not react with each components alpha 5, beta 1, gamma 1 and similar isoform laminin-511.
Note: Laminin 511 is used as a culture substrate for ES and iPS cells.
Fig.2 Structure of Laminin-511
|Description||Anti-Laminin alpha 3A||Anti-Laminin alpha 3B||Anti-Laminin gamma 2 N-terminal Fragment||Anti-Laminin-511|
|Specificity||Laminin alpha 3A *1|
|Host and Type||Mouse Monoclonal|
|Reactivity||Human||Human||Human and Mouse||Human|
|Form||Mouse Ascites (Unpurified)|
|Application||ELISA, IHC, IP, WB||IHC (frozen & paraffin sextions), WB||IHC (frozen), IP, WB (non-reducing) *2|
*1 Clone BG5 only detects alpha 3A in IHC. However, this clone may react with alpha 3B in ELISA, IP and Western Blot.
*2 Clone 12D can be only used for IHC (Frozen) and under non-reducing condition for Western Blot.
*3 Non-preservative added. So these 4 clones can be degraded at 4C. We recommend -20C storage.
Fig.3 IHC image of skin cancer tissue in clone BG5 and F7
Left : Clone BG5, Right : Clone F7
Both clones detectedbasal membrane like structure around the tumor but F7 clone also detected
vascular basement membrane (arrow heads).
Fig.4 IHC image of tissues in clone P2H
Upper images : Normal mammary gland
Lower images: Breast cancer
P2H stains basement membranes surrouding mammary glands in normal tisue,
whereas in a breast cancer tissue cytoplasmic accumulation of γ2pf orits fragments
Fig.5 IHC image ofbreast cancer tissue in clone 12D
A : Vascular basement membrane
B: Mammary gland basement membrane
C : Basement membrane-like structures surrounding tumor cells
Fig.6 Specificity of clone 12D against laminin isoforms
Seven recombinant human laminins were coated on a multi-well
plate and detected by clone 12D. The data shows clone 12D
specifically reacts with Laminin 511.
Citations and References