Mouse cortical glia stained with anti-Iba1 (red) and Hoechst (blue). Image provided by Simon Glerup, Aarhus University.
Ghatak et al. MethodsX 2014 show that Iba-1 signal using this antibody is increased following antigen retrieval using EDTA, pH 6.0.
See list of additional Iba1 antibodies at Linscott’s Directory
On July 6, 2020 Jamie Rose, University of Edinburgh wrote:
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#!
On May 13, 2020 Dimitri Traenkner, University of Utah wrote:
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
On April 16, 2020 Johannes Schlachetzki wrote:
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
On April 29, 2017 Pall Karlson, Department of Clinical Medicine, Aarhus University wrote:
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.
On May 22, 2015 Jean-Martin Lapointe wrote:
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.
On September 28, 2014 Simon Glerup, Mayo Clinic wrote:
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
On March 6, 2014 Christian Vægter wrote:
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.
On May 20, 2012 Simon G, Aarhus wrote:
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.