Anti CX3CR1, Guinea Pig

CX3CR1 is a type of G protein-coupled receptor (GPCR) for the fractalkine CX3CL1. It is known to be involved in cell survival, migration and proliferation. It is expressed on macrophages and microglia and is used as a marker for these cells. It has been reported that in CX3CR1 deficient mice, synaptic pruning by microglia is not sufficient, resulting in abnormal synaptic transmission between neurons, and it is thought to be required for synaptic pruning¹⁾. The “Anti CX3CR1, Guinea Pig” is a guinea pig polyclonal antibody, raised against CX3CR1^2-3). 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 (24 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

Primary Antibody:

Iba1: Anti Iba1, Rabbit (for Immunocytochemistry) (Product No. 019-19741) 1:500
CX3CR1: Anti CX3CR1, Guinea Pig (This product) 1:800

Neurons were detected using NeuroTrace™ 435/455 Blue Fluorescent Nissl Stain (ThermoFisher Scientific)

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

1. Zhan, Y. et al.: Nat. Neurosci., 17(3), 400(2014).
   Deficient neuron-microglia signaling results in impaired functional brain connectivity and social behavior
2. Takagi, S. et al.: J. Neuroimmunology, 331, 74(2019).
   Microglia are continuously activated in the circumventricular organs of mouse brain
3. Kawai, S. et al.: Cell Biochem. Funct., 38(4), 392(2020).
   Transient increase of microglial C1q expression in the circumventricular organs of adult mouse during LPS‐induced inflammation