Anti-Laminin γ2 N-terminal fragment, Human, Mouse-Mono (Clone P2H)

Product#: FDV-0025
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Anti-Laminin γ2 N-terminal fragment, Human, Mouse-Mono (Clone P2H)


General information 

Cat. No. FDV-0025
Size 100 ul
Format Mouse ascites
Formulation Ascites without any additives
Host Species and Clonality Mouse monoclonal
Isotype and Subclass  IgG2a
Purification No purification
Lot Number see vial label
Specificity Human, other species not tested yet
Antigen recombinant human γ2pf containing domain IV + V
Epitope γ2 domain V, NE2
Application  
  1. Western blotting under both reducing, and non-reducing conditions (1/1,000-1/10,000)
  2. Immunohistochemistry with paraffin and frozen sections
  3. ELISA
Storage For short-term storage, -20oC. For long-term storage, -80oC storage is preferable. 
    Avoid repeated freeze-thaw cycles and avoid storage at 4oC


Description

Laminins, which consist of three subunits called α, β and γ chains, are major cell-adhesive components of extracellular matrix, especially basement membranes (BMs). The laminin family is constituted of over 15 isoforms, expressed in a tissue-specific manner and plays differential roles in each tissue. Laminin γ2 chain is a subunit of laminin-332 isoform, formerly laminin-5, but it is frequently overexpressed as a monomer form or the β3-γ2 heterodimer in invasive cancers. Laminin γ2 is a protein of approximately 150 kDa and cleaved at its short arm mainly by bone morphogenic protein-1 (BMP1) in cancer cells, releasing an N-terminal proteolytic fragment (γ2pf) of 45 kDa. The short arm and γ2pf are also cleaved by MMPs and serine proteinases, releasing further small N-terminal fragments. Recently, Dr. Kaoru Miyazaki and co-workers generated a novel mouse monoclonal antibody against γ2pf, clone P2H, which can efficiently detect invasive cancer cells. Clone P2H recognizes laminin γ2 chain of laminin-332 on normal epithelial basement membranes, while in malignant cancer tissues it strongly immunostains the cytoplasm of cancer cells at invasion fronts. Thus, P2H is an important antibody capable of highly detecting the tumor invasion marker laminin γ2 chain. P2H is an only commercially available antibody to detect the N-terminal proteolytic fragments of laminin γchain in cancer tissues. It is useful for pathological assessment of invasiveness and malignancy in various types of human cancers. P2H is applicable to not only immunohistochemistry, but also immunoblotting, ELISA and immunoprecipitation.
 
F9.png
Figure 1. Protein structure of laminin-3A32s and cleavage site/N-terminal fragment of γ2 chain

Application examples
 
Western blot
Samples :
(left) recombinant human γ2pf under reducing conditions
(right) conditioned medium of VMRC-LCP lung cancer cells. Small γ2 N-terminal fragments are seen below the γ2pf band (45 kDa), and 15-kDa bands corresponds to NE1-2 and NE2-3.

Gel conc. : 10% (left), gradient gel (right)
1 st Antibody : clone P2H, 1/1,000 dilution
2 nd Antibody : anti-mouse IgG (H+L)-HRP conjugate
F10.png
Immunohistochemistry : Normal mammary gland and breast cancer
Upper panel : Normal mammary gland
Lower panel : Breast cancer

P2H stains basement membranes surrounding mammary glands in normal tissue, whereas in a breast cancer tissue cytoplasmic accumulation of γ2pf or its fragments is detected with P2H.
F11.png
Immunohistochemistry : Normal skin and skin cancer
Upper panel : Normal skin tissue
Lower panel : Skin cancer tissue

Although P2H stains epithelial basement membranes in normal skin, in a skin cancer tissue it strongly stains invading cancer cells or their cytoplasm.
F12.png
Immunohistochemistry : Lung cancers - SCC???????
Right upper panel : Stained by P2H (anti-γ2pf)
Right lower panel : Stained by BG5 (anti-α3A, cat. No. FDV-0024)

Although BG5 detected basement membrane (BM)-like structure surrounding tumor cells, P2H detected not only BMs but also the cytoplasm of cancer cells.
F13.png
Immunohistochemistry : Lung SCC and ADC
Upper two panels : Invasive lung SCC
Lower two panels : Invasive lung ADC

P2H strongly detected cytoplasm of cancer cells at invasion fronts.
F14.png

Reference
  1. Miyazaki et al., Cancer Sci., 107, 1909-1918 (2016) Highly sensitive detection of invasive lung cancer cells by novel antibody against amino-terminal domain of laminin gamma2 chain.

Related products
 

Clone

BG5

F7

P2H

12D

Target

Laminin-α3A chain

Laminin-α3B chain

Laminin-γ2 chain N-terminal region

Laminin-511 trimer

Normal Tissue

FDV-0023_1-1.jpg

FDV-0023_1-2.jpg

FDV-0023_1-3.jpg

FDV-0023_1-4.jpg

Epitope

FDV-0023_2-1.jpg

FDV-0023_2-2.jpg

FDV-0023_2-3.jpg

FDV-0023_2-4.jpg

Cancer Tissue

FDV-0023_3-1.jpg
 

FDV-0023_3-2.jpg
 

FDV-0023_3-3.jpg
 

FDV-0023_3-4.jpg
 


Outline of four clones
 

Description

Anti-Laminin alpha 3A

Anti-Laminin alpha 3B

Anti-Laminin gamma 2 N-terminal Fragment

Anti-Laminin-511

Clone

BG5

F7

P2H

12D

Specificity

Laminin alpha 3A*1

Laminin alpha 3B

Laminin gamma 2 Domain V-NE2

Laminin-511

Host and Type

Mouse Monoclonal

Subclass

IgG2a

IgG1

IgG2a

IgG2b

Reactivity

Human

Human

Human and Mouse

Human

Form

Mouse Ascites (Unpurifed)

     

Application

ELISA, IHC, IP, WB

IHC (frozen & paraffin sections), WB

IHC (frozen), IP, WB Non-reducing) *2

Code

FDV-0024

FDV-0023

FDV-0025

FDV-0026

Size

100 uL

Storage

-20oC *3


*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 4oC. We recommend -20oC storage.
 

Aliases for LAMC2 Gene

  • Laminin Subunit Gamma 2 2 3 5
  • Large Adhesive Scatter Factor 140 KDa Subunit 3 4
  • Cell-Scattering Factor 140 KDa Subunit 3 4
  • Laminin Subunit Gamma-2 3 4
  • Epiligrin Subunit Gamma 3 4
  • Ladsin 140 KDa Subunit 3 4
  • Kalinin Subunit Gamma 3 4
  • Nicein Subunit Gamma 3 4
  • CSF 140 KDa Subunit 3 4
  • Laminin B2t Chain 3 4
  • Laminin, Gamma 2 2 3
  • BM600-100kDa 2 3
  • LAMB2T 3 4
  • LAMNB2 3 4
  • Laminin, Gamma 2 (Nicein (100kD), Kalinin (105kD), BM600 (100kD), Herlitz Junctional Epidermolysis Bullosa)) 2
  • Kalinin/Nicein/Epiligrin 100 KDa Subunit 4
  • Laminin-5 Subunit Gamma 4
  • Kalinin-105kDa 2
  • Nicein-100kDa 2
  • BM600 3
  • EBR2A 3
  • LAMC2 5
  • EBR2 3
  • B2T 3
  • CSF 3
 

 

 
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