Revision 5
Cell Signaling Technology

Orders: 877-616-CELL (2355) orders@cellsignal.com

Support: 877-678-TECH (8324)

Web: info@cellsignal.com cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP, IHC-Bond, IHC-P, IF-F, IF-IC, FC-FP

REACTIVITY:

H M R Hm Mk

SENSITIVITY:

Endogenous

MW (kDa):

17

Source/Isotype:

Rabbit IgG

UniProt ID:

#P55008

Entrez-Gene Id:

199

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
IHC Leica Bond 1:800 - 1:3200
Immunohistochemistry (Paraffin) 1:800 - 1:3200
Immunofluorescence (Frozen) 1:50 - 1:200
Immunofluorescence (Immunocytochemistry) 1:50 - 1:200
Flow Cytometry (Fixed/Permeabilized) 1:50 - 1:200

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

For a carrier-free (BSA and azide free) version of this product see product #79394.

Specificity / Sensitivity

Iba1/AIF-1 (E4O4W) XP® Rabbit mAb recognizes endogenous levels of total Iba1/AIF-1 protein.

Species Reactivity:

Human, Mouse, Rat, Hamster, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala139 of human Iba1/AIF-1 protein.

Background

Ionized calcium-binding adaptor molecule 1 (Iba1), also known as allograft inflammatory factor 1 (AIF-1), is an evolutionarily conserved cytoplasmic calcium binding protein containing a central pair of EF-hand calcium binding motifs (1,2). Iba1/AIF-1 was originally cloned from activated macrophages in human atherosclerotic allogenic heart grafts undergoing chronic transplant rejection as well as from rat monocytes (3,4). Its function is not very well understood, but Iba1/AIF-1 expression is upregulated in response to interferon-gamma and, therefore, could modulate macrophage-dependent immune response (3). As an F-actin-binding protein, Iba1/AIF-1 may function to remodel the actin cytoskeleton and contribute to morphological changes that correlate with various microglial/macrophage states (5). Iba1/AIF-1 is also uniquely expressed in cells of monocytic lineage and is, therefore, widely used as a marker for microglia/macrophages in the brain and other tissue (6).

  1. Schulze, J.O. et al. (2008) FEBS J 275, 4627-40.
  2. Deininger, M.H. et al. (2002) FEBS Lett 514, 115-21.
  3. Utans, U. et al. (1995) J Clin Invest 95, 2954-62.
  4. Imai, Y. et al. (1996) Biochem Biophys Res Commun 224, 855-62.
  5. Kanazawa, H. et al. (2002) J Biol Chem 277, 20026-32.
  6. Ito, D. et al. (1998) Brain Res Mol Brain Res 57, 1-9.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation IHC-Bond: IHC Leica Bond IHC-P: Immunohistochemistry (Paraffin) IF-F: Immunofluorescence (Frozen) IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Revision 5
#17198

Iba1/AIF-1 (E4O4W) XP® Rabbit mAb

Western Blotting Image 1: Iba1/AIF-1 (E4O4W) XP® Rabbit mAb Expand Image
Western blot analysis of extracts from various cell lines using Iba1/AIF-1 (E4O4W) XP® Rabbit mAb (upper) and β-Tubulin (D2N5G) Rabbit mAb #15115 (lower).
Immunoprecipitation Image 1: Iba1/AIF-1 (E4O4W) XP® Rabbit mAb Expand Image
Immunoprecipitation of Iba1/AIF-1 from THP-1 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Iba1/AIF-1 (E4O4W) XP® Rabbit mAb. Western blot analysis was performed using Iba1/AIF-1 (E4O4W) XP® Rabbit mAb.
Immunohistochemistry Image 1: Iba1/AIF-1 (E4O4W) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human neuroendocrine carcinoma of the lung using Iba1/AIF-1 (E4O4W) XP® Rabbit mAb performed on the Leica® BOND Rx.
Immunohistochemistry Image 2: Iba1/AIF-1 (E4O4W) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human non-Hodgkin's lymphoma using Iba1/AIF-1 (E4O4W) XP® Rabbit mAb performed on the Leica® BOND Rx.
Immunohistochemistry Image 3: Iba1/AIF-1 (E4O4W) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human ovarian clear cell carcinoma using Iba1/AIF-1 (E4O4W) XP® Rabbit mAb (left) or Iba1/AIF-1 (E5N4J) Mouse mAb (IHC Formulated) #58970 (right) performed on the Leica® BOND Rx. These two antibodies detect independent, unique epitopes on human Iba1/AIF-1 protein. The similar staining patterns obtained with both antibodies help to confirm the specificity of the staining.
Immunohistochemistry Image 1: Iba1/AIF-1 (E4O4W) XP® Rabbit mAb Expand Image
Dual immunohistochemical analysis of paraffin-embedded human Alzheimer's brain using Iba1/AIF-1 (E4O4W) XP® Rabbit mAb (brown) and APP/β-Amyloid (NAB228) Mouse mAb #2450 (red).
Immunohistochemistry Image 2: Iba1/AIF-1 (E4O4W) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded mouse brain using Iba1/AIF-1 (E4O4W) XP® Rabbit mAb.
Immunohistochemistry Image 3: Iba1/AIF-1 (E4O4W) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded mouse spleen using Iba1/AIF-1 (E4O4W) XP® Rabbit mAb.
Immunohistochemistry Image 4: Iba1/AIF-1 (E4O4W) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded mouse small intestine using Iba1/AIF-1 (E4O4W) XP® Rabbit mAb.
Immunohistochemistry Image 5: Iba1/AIF-1 (E4O4W) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded normal rat brain using Iba1/AIF-1 (E4O4W) XP® Rabbit mAb.
Immunohistochemistry Image 6: Iba1/AIF-1 (E4O4W) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded normal rhesus monkey spleen using Iba1/AIF-1 (E4O4W) XP® Rabbit mAb.
Immunohistochemistry Image 7: Iba1/AIF-1 (E4O4W) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded normal rhesus monkey liver using Iba1/AIF-1 (E4O4W) XP® Rabbit
Immunohistochemistry Image 8: Iba1/AIF-1 (E4O4W) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded normal Syrian hamster small intestine using Iba1/AIF-1 (E4O4W) XP® Rabbit mAb.
Immunohistochemistry Image 9: Iba1/AIF-1 (E4O4W) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human ovarian serous carcinoma using Iba1/AIF-1 (E4O4W) XP® Rabbit mAb.
Immunohistochemistry Image 10: Iba1/AIF-1 (E4O4W) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Iba1/AIF-1 (E4O4W) XP® Rabbit mAb.
Immunohistochemistry Image 11: Iba1/AIF-1 (E4O4W) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human ductal breast carcinoma using Iba1/AIF-1 (E4O4W) XP® Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
Immunohistochemistry Image 12: Iba1/AIF-1 (E4O4W) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded THP-1 cell pellet (left, positive) or SH-SY5Y cell pellet (right, negative) using Iba1/AIF-1 (E4O4W) XP® Rabbit mAb.
Immunofluorescence Image 1: Iba1/AIF-1 (E4O4W) XP® Rabbit mAb Expand Image
Confocal immunofluorescent analysis of human cortex (left) and mouse CA1 hippocampus (right) using Iba1/AIF-1 (E4O4W) XP® Rabbit mAb (green). In mouse tissue sections, cell nuclei were labeled with DAPI (blue). Images kindly provided by Dr. Simone Brioschi and Dr. Marco Colonna (Washington University) and used with permission.
Immunofluorescence Image 2: Iba1/AIF-1 (E4O4W) XP® Rabbit mAb Expand Image
Confocal immunofluorescent analysis of mouse small intestine using Iba1/AIF-1 (E4O4W) XP® Rabbit mAb (green). Actin filaments were labeled with DyLight 554 Phalloidin #13054 (red). Sections were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Immunofluorescence Image 3: Iba1/AIF-1 (E4O4W) XP® Rabbit mAb Expand Image
Confocal immunofluorescent analysis of mouse liver using Iba1/AIF-1 (E4O4W) XP® Rabbit mAb (green). Actin filaments were labeled with DyLight 554 Phalloidin #13054 (red). Sections were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Immunofluorescence Image 4: Iba1/AIF-1 (E4O4W) XP® Rabbit mAb Expand Image
Confocal immunofluorescent analysis of microglia in mouse hippocampus using Iba1/AIF-1 (E4O4W) XP® Rabbit mAb (green). Sections were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Immunofluorescence Image 1: Iba1/AIF-1 (E4O4W) XP® Rabbit mAb Expand Image
Confocal immunofluorescent analysis of THP-1 cells differentiated with TPA (12-O-Tetradecanoylphorbol-13-Acetate) #4174 (80 nM, 24 hr; left, positive) and SH-SY5Y cells (right, negative), using Iba1/AIF-1 (E4O4W) XP® Rabbit mAb (green). Actin filaments were labeled with DyLight 554 Phalloidin #13054 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Flow Cytometry Image 1: Iba1/AIF-1 (E4O4W) XP® Rabbit mAb Expand Image
Flow cytometric analysis of SH-SY5Y cells (blue, negative) and THP-1 cells (green, positive) using Iba1/AIF-1 (E4O4W) XP® Rabbit mAb (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.