Anti-Iba-1 Polyclonal Rabbit Antibody from Wako

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Donnelly Center for Cellular and Biomolecular Research
University of Toronto
Postdoctoral Researcher
Brain tissue section (frozen, 5 uM) from embryonic (E14) mouse brain stained with anti- Iba-1 antibody (1:1000) shows ramified microglia with distinct processes.

Company:

Wako Chemicals USA

Product Name:

Anti-Iba-1 Polyclonal Antibody, Rabbit

Catalog Number:

019-19741

Image

For the detection of microglia in mouse and human brain and spinal cord tissues, Iba-1 has been identified as a good marker candidate. Indeed, Iba-1 is specifically expressed in macrophages/microglia and hence using this marker, one can clearly distinguish microglia/macrophages from astrocytes and neurons in the nervous system. The most useful purpose of the Anti-Iba-1 antibody is to differentiate normal quiescent microglia from activated and inflammatory microglia. This is important from the standpoint of neuroinflammation which was my area of research. Specifically, I also wanted to identify how microglia contribute to the growth and development of astrocytes and neurons in the developing embryo and during developmental neuropathogenesis (Antony JM, 2011a,b) .

Experimental Design and Results Summary

Applications

Immunohistochemistry (IHC), immunocytochemistry (ICC), fluorescence activated cell sorting (FACS)

Sample

Frozen and paraffin sections and primary cultures from embryonic and postnatal mouse brain tissue

Primary Incubation

Iba-1 4°C overnight incubation at 1:1000 in 3% normal goat serum (NGS) and 0.26% bovine serum albumin (BSA) in hepes buffered saline (HBS)

Blocking Agent

6% NGS, 0.5% BSA in HBS, 1 hour at room temperature.

Secondary Incubation

Alexa488-conjugated goat anti-rabbit (Jackson Immunoresearch, Cat # 111-545-003) at 1:10,000 in 3% NGS and 0.26% BSA in HBS, for 1 hour at room temperature

Tertiary Incubation

N/A

Detection

Fluorescence (Zeiss Axioplan2 upright microscope with SPOT2 camera)

Results Summary

Iba-1 is exclusively present on microglia in embryonic and postnatal mouse brain tissue. The signal is very strong with no background signal in any other cell type. Poor washing in PBS can lead to low level detection of Iba-1 in other cell types.

Additional Notes

Iba-1 as a marker has been well established and has shown consistent results in several experiments. Indeed, Iba-1 transgenic mice are available (Hirasawa et al., 2005).

Image Gallery

Brain tissue section (frozen, 5 uM) from embryonic (E14) mouse brain stained with anti- Iba-1 antibody (1:1000) shows ramified microglia with distinct processes.

Summary

The Good

Anti-Iba-1 polyclonal antibody can be used for several applications (immunocytochemistry and immunohistochemistry, FACS)

The Bad

None.

The Bottom Line

Good antibody and probably the best antibody currently available for detection of microglia.

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