Iba1, Fujifilm Wako Pure Chemical Corporations, 019-19741

Great for IF and ICH. 1:1000 with 2% EDTA retrieval.

For more information see Pickett et al. Cell Reports 2019, “Amyloid Beta and Tau Cooperate to Cause Reversible Behavioral and Transcriptional Deficits in a Model of Alzheimer’s Disease”.
https://www.sciencedirect.com/science/article/pii/S2211124719315268?dgcid=rss_sd_all#!

This antibody works roughly for IHC.

For more information see Tränkner et al. Cell Reports 2019, “A Microglia Sublineage Protects from Sex-Linked Anxiety Symptoms and Obsessive Compulsion”
https://www.sciencedirect.com/science/article/pii/S2211124719312331

This antibody worked great for both IHC and IF (mouse).
Dilution used: 1:500

For more information see Süß et al. Cell Reports 2020, “Chronic Peripheral Inflammation Causes a Region-Specific Myeloid Response in the Central Nervous System”.
https://www.sciencedirect.com/science/article/pii/S2211124720302904

Anti-Iba1, rabbit polyclonal antibody from Wako Chemicals (Cat. No. – 019-19741)

Used on frozen 50-micrometer thick human skin sections.
Worked quite well in 1:500 dilution over night (immunofluorescence) with positive stainings both in epidermis and dermis of the skin.

The Iba1 antibody is very robust in formalin-fixed paraffin-embedded tissue. It stains not only microglia but multiple macrophage-type cells in many tissues (intestinal lamina propria macrophages, Kupffer cells in liver, splenic red pulp macrophages, etc…). We use heat retrieval (pressure cooker) in pH6 buffer, incubate the primary for 1 hour at room temperature (around 1:5000 dilution), and use an HRP-linked secondary and DAB chromogen. There can be some punctate non-specific staining in nuclei, but you can get rid of most of it by diluting the antibody down.

The image above shows selective Iba1 staining of primary mouse cortical glia. The signal seems very specific but one could wish for higher sensitivity.
Cells were permeabilized in PBS+0.1% TritonX100 and blocked in PBS+10% FBS.
Dilution for IF 1:500

Used for IF on cultured murine microglia, 1:500. Worked very well and seems specific for microglia. Also used on spinal cord sections, equally well, with clear increase in Iba1+ cells ipsilateral to sciatic injury.

This Iba1 antibody is a great marker for microglia.
It specifically stains these and not other glia in PFA fixed cryosections of mouse brain.
It does however faintly stain nuclei of neurons in our hands.