Revision 6

#17198

Store at -20C

CST Logo
Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Applications:
W, W-S, 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 Usage Information

Application Dilution
Western Blotting 1:1000
Simple Western™ 1:10 - 1:50
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.

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).

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

W: Western Blotting W-S: Simple Western™ 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

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.

限制使用

除非 CST 的合法授书代表以书面形式书行明确同意,否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件,除非书 CST 的合法授书代表以书面形式书独接受, 否书均被拒书,并且无效。

专品专有“专供研究使用”的专专或专似的专专声明, 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的, 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专,且专用于研专用途。将专品用于专断、专防或治专目的, 或专专售(专独或作专专成)或其他商专目的而专专专品,均需要 CST 的专独专可。客专:(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品,或使用专品制造任何商专专品,(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专,或使用专品开专任何与 CST 的专品或服专专争的专品或服专, (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专,(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品 , (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 6

Cell Signaling Technology Logo
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).
Western Blotting Image 1: Iba1/AIF-1 (E4O4W) XP<sup>®</sup> Rabbit mAb
Simple Western™ analysis of lysates (1 mg/mL) from THP-1 cells using Iba1/AIF-1 (E4O4W) XP® Rabbit mAb #17198. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
Western Blotting Image 1: Iba1/AIF-1 (E4O4W) XP<sup>®</sup> Rabbit mAb
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.
Immunoprecipitation Image 1: Iba1/AIF-1 (E4O4W) XP<sup>®</sup> Rabbit mAb
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 6

Cell Signaling Technology Logo
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 1: Iba1/AIF-1 (E4O4W) XP<sup>®</sup> Rabbit mAb
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 2: Iba1/AIF-1 (E4O4W) XP<sup>®</sup> Rabbit mAb
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 3: Iba1/AIF-1 (E4O4W) XP<sup>®</sup> Rabbit mAb
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 6

Cell Signaling Technology Logo
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 1: Iba1/AIF-1 (E4O4W) XP<sup>®</sup> Rabbit mAb
Immunohistochemical analysis of paraffin-embedded mouse brain using Iba1/AIF-1 (E4O4W) XP® Rabbit mAb.
Immunohistochemistry Image 2: Iba1/AIF-1 (E4O4W) XP<sup>®</sup> Rabbit mAb
Immunohistochemical analysis of paraffin-embedded mouse spleen using Iba1/AIF-1 (E4O4W) XP® Rabbit mAb.
Immunohistochemistry Image 3: Iba1/AIF-1 (E4O4W) XP<sup>®</sup> Rabbit mAb
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 6

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded mouse small intestine using Iba1/AIF-1 (E4O4W) XP® Rabbit mAb.
Immunohistochemistry Image 4: Iba1/AIF-1 (E4O4W) XP<sup>®</sup> Rabbit mAb
Immunohistochemical analysis of paraffin-embedded normal rat brain using Iba1/AIF-1 (E4O4W) XP® Rabbit mAb.
Immunohistochemistry Image 5: Iba1/AIF-1 (E4O4W) XP<sup>®</sup> Rabbit mAb
Immunohistochemical analysis of paraffin-embedded normal rhesus monkey spleen using Iba1/AIF-1 (E4O4W) XP® Rabbit mAb.
Immunohistochemistry Image 6: Iba1/AIF-1 (E4O4W) XP<sup>®</sup> Rabbit mAb
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 6

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded normal rhesus monkey liver using Iba1/AIF-1 (E4O4W) XP® Rabbit
Immunohistochemistry Image 7: Iba1/AIF-1 (E4O4W) XP<sup>®</sup> Rabbit mAb
Immunohistochemical analysis of paraffin-embedded normal Syrian hamster small intestine using Iba1/AIF-1 (E4O4W) XP® Rabbit mAb.
Immunohistochemistry Image 8: Iba1/AIF-1 (E4O4W) XP<sup>®</sup> Rabbit mAb
Immunohistochemical analysis of paraffin-embedded human ovarian serous carcinoma using Iba1/AIF-1 (E4O4W) XP® Rabbit mAb.
Immunohistochemistry Image 9: Iba1/AIF-1 (E4O4W) XP<sup>®</sup> Rabbit mAb
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 6

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Iba1/AIF-1 (E4O4W) XP® Rabbit mAb.
Immunohistochemistry Image 10: Iba1/AIF-1 (E4O4W) XP<sup>®</sup> Rabbit mAb
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 11: Iba1/AIF-1 (E4O4W) XP<sup>®</sup> Rabbit mAb
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.
Immunohistochemistry Image 12: Iba1/AIF-1 (E4O4W) XP<sup>®</sup> Rabbit mAb
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 6

Cell Signaling Technology Logo
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 1: Iba1/AIF-1 (E4O4W) XP<sup>®</sup> Rabbit mAb
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 2: Iba1/AIF-1 (E4O4W) XP<sup>®</sup> Rabbit mAb
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 3: Iba1/AIF-1 (E4O4W) XP<sup>®</sup> Rabbit mAb
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 6

Cell Signaling Technology Logo
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 4: Iba1/AIF-1 (E4O4W) XP<sup>®</sup> Rabbit mAb
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).
Immunofluorescence Image 1: Iba1/AIF-1 (E4O4W) XP<sup>®</sup> Rabbit mAb
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.
Flow Cytometry Image 1: Iba1/AIF-1 (E4O4W) XP<sup>®</sup> Rabbit mAb
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.