Anti Laminin, Guinea Pig

Laminin is a glycoprotein with a molecular weight of approximately 900 kDa. It consists of three chains-α, β, and γ-that assemble into a cross-shaped structure. Laminin is well-known as a key component of the extracellular matrix (ECM) and plays an important role in cell adhesion, migration, and proliferation. In the central nervous system, laminin is almost exclusively expressed in the basement membranes surrounding blood vessels and is used as a marker for these vessels. Previous studies have shown that the inner (endothelial) layer of the basement membrane in blood vessels contains laminin-411 and -511, while the outer (parenchymal) layer contains laminin-211 and -111¹⁾. Laminin also plays a critical role in the formation and function of the blood-brain barrier (BBB).

The “Anti Laminin , Guinea Pig” is a guinea pig polyclonal antibody, raised against Laminin^2-6). It can be used to perform multiplex immunohistochemistry.

Antibody Information

Protocol (Example)

1.Perfusion fixation

Perfuse with sodium citrate/PBS to remove blood, followed by perfusion with 4% PFA.

2.Post-fixation

4% PFA (Recommended time: 6 hours)

3.Immersion in sucrose

30% Sucrose/PBS (4℃, from overnight to approximately 2 days)

4.Preparation of frozen sections

After removing sucrose, prepare frozen blocks and cut 30 μm thick sections using a cryostat.

5.Glycine treatment

25 mM glycine/PBS (Approximately 20 minutes)

6.Blocking

5% normal goat serum in 0.3% Triton X-100/PBS (4℃, Overnight)

7.Primary antibody reaction

Anti Laminin, Guinea pig (1:500), 1% normal goat serum in 0.3% Triton X-100/PBS (4℃, from overnight to 2 days).

8.Washing

0.3% TritonX-100/PBS (5 minutes×3)

9.Secondary antibody reaction

Alexa Fluor^® 488 AffiniPure Goat Anti-Guinea Pig IgG (H+L) (1:500), 1% normal goat serum in 0.3% Triton X-100/PBS (RT, 2 hours)

10.Washing

0.3% TritonX-100/PBS (5 minutes×3)

11.Mounting

The sample is sealed in mounting medium and stored at 4℃ in a dark place.

Application Data

Immunohistochemistry

Species: Mouse
Site: Hippocampus
Sample: Frozen section
Antibody concentration: 1:500

Data by courtesy of
Dr. Miyata, Department of Applied Biology, Kyoto Institute of Technology

Species: Mouse
Site: Medulla oblongata (Area postrema)
Sample: Frozen section
Antibody concentration: 1:500

Data by courtesy of
Dr. Miyata, Department of Applied Biology, Kyoto Institute of Technology

1. Halder, S. K. et al.: Neural Regen. Res., 18(12), 2557(2023).
   The importance of laminin at the blood-brain barrier
2. Morita, S. et al.: Cell Tissue Res., 363, 497(2016).
   Heterogeneous vascular permeability and alternative diffusion barrier in sensory circumventricular organs of adult mouse brain
3. Nishikawa, K. et al.: J. Neuroendocrinol., 29(2), (2017).
   Structural Reconstruction of the Perivascular Space in the Adult Mouse Neurohypophysis During an Osmotic Stimulation
4. Takagi, S. et al.: J. Neuroimmunol., 331, 74(2019).
   Microglia are continuously activated in the circumventricular organs of mouse brain
5. Muneoka, S. et al.: J. Neuroimmunol., 331, 58(2019).
   TLR4 in circumventricular neural stem cells is a negative regulator for thermogenic pathways in the mouse brain
6. Murayama, S. et al.: J. Neuroimmunol., 334, 576973(2019).
   Activation of microglia and macrophages in the circumventricular organs of the mouse brain during TLR2-induced fever and sickness responses