2. Antibodies

image
Figure:
Immuno-double staining of rat primary mixed culture cells

Green:
Iba1, which reacts to anti Iba1 antibody (Wako Cat. #019-19741)

Red:
astrocyte, which reacts to anti GFAP, monoclonal antibody

(Data was provided by Dept. of Neurochemistry, National Institute of Nueroscience (Japan).)
Antibodies against Macrophage/ Microglia-specific Protein Iba1*

* Iba1: ionized calcium binding adapter molecule

Anti Iba-1 polyclonal antibody, Rabbit, for Immunocytochemistry
Wako Cat. No. 019-19741 (50 µg (100 µL)) <for Immunochemistry>
Keep at -20°C
Working Concentration: 1 ~ 2 µg/mL (Immunocytochemistry)
Anti Iba-1 polyclonal antibody, Rabbit, for Western Blotting
Wako Cat. No. 016-20001 (50 µg (100 µL)) <for Immunochemistry>
Keep at -20°C
Working Concentration: 0.5 ~ 1 µg/mL (Western Blot)
Calcium ions are known to be one of the most important signal mediators in all cells including
central nervous system (CNS) cells. Calcium ions exert their signaling activity through
association with various calcium binding proteins, many of which are classified into a large
protein family, the EF hand protein family.
Iba1 is a 17-kDa EF hand protein that is specifically expressed in macrophages/ microglia
and is upregulated during the activation of these cells.
Wako distributes rabbit polyclonal antibodies were raised against a synthetic peptide
corresponding to the Iba1 carboxy-terminal sequence, which was conserved among human,
rat and mouse Iba1 protein sequences. These antibodies are specifically reactive to
microglia/ macrophages, are appropriate for immuno-double staining of brain tissues and cell
culture in combination with monoclonal antibody to GFAP, which specifically reacts to astrocyte.


[Specificity]
Specific to microglia and macrophages, but not cross-reactive with neurons and astrocytes. Reactive with human, mouse and rat Iba1.
image
Figure :
Immunofl uorescence staining of adult mouse olfactory epithelium with goat anti-OMP (Wako Cat. # 544-10001).

Green :
OMP staining was visualized with Cy2 (Jackson Immuno Research).

Data was provided by Dr. Frank L. Margolis and Dr. Jae Hyung Koo, Department of Anatomy and Neurobiology, School of Medicine, University of Maryland.
Research for Olfactory Nerve
Anti Olfactory Marker Protein, Goat [Anti OMP]
Wako Cat. No. 019-22291* (100 µL) <for Immunochemistry>
Keep at -20°C
*: The catalog number was used to be 544-10001.
Olfactory Marker Protein (OMP) is soluble acid protein expressed in mature olfactory nerve. This goat antiserum is highly specifi c for mature olfactory neurons and their axons and terminals in tissue sections of many vertebrate species including rodents, humans, marsupials and amphibia.
Working Dilutions:
Western Blot: ~ 1 : 50,000
Immunocytochemistry: 1 : 200 (paraffin embedded material)
1 : 50,000 (Vectastain-Elite with fixed floating sections)
Preparation:
Goat antiserum to OMP (100 μL) is diluted 1:1 with glycerol containing 0.05% sodium azide to facilitate shipment at ambient temperature.
[References]
1) Baker, H. et al. : J. Comp. Neurol., 285, 246 (1989).
2) Buiakova, O. I. et al. : Genomics, 20, 452 (1994).
3) Cummings, D. M. et al. : J. Comp. Neurol., 421, 362 (2000).
4) Keller, A. and Margolis, F. L. : J. Neurochem., 24, 1101 (1975).
5) Koo, J. H. et al. : J. Neurochem., 90, 102 (2004).
6) Koo, J. H. et al. : J. Comp. Neurol., 487, 1 (2005).
7) Rama Krishna, N. S. et al. : Brain Res., 593, 295 (1992).
8) Verhaagen, J. et al. : J. Neurosci. Res., 26, 31 (1990).
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Wako Product Update Bio-No.1 [ page. 7 ]