Revision 6
Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

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

WB, 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 Information

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.

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 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 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.

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

Revision 6
#17198

Iba1/AIF-1 (E4O4W) XP® Rabbit mAb

Western Blotting Image 1: Iba1/AIF-1 (E4O4W) XP® Rabbit mAb Expand Image
使用 Iba1/AIF-1 (E4O4W) XP® Rabbit mAb(上图)和 β-Tubulin (D2N5G) Rabbit mAb #15115 (下图)对不同细胞系的提取物进行蛋白印迹分析。
Western Blotting Image 1: Iba1/AIF-1 (E4O4W) XP® Rabbit mAb Expand Image
Simple Western™ analysis of lysates (1 mg/mL) from THP-1 cells using Iba1/AIF-1 (E4O4W) XP® Rabbit mAb #17198. 虚拟泳道式图像(左图)显示一抗稀释比例为 1:10 和 1:50 时的靶标条带(如图所示)。对应的电泳图(右图)为一抗稀释比例在 1:10(蓝线)和 1:50(绿线)时沿毛细血管内分子量的化学发光结果。在还原条件下,使用 12-230 kDa 分离模块在 ProteinSimple(BioTechne 品牌)的 Jess™ Simple Western 仪器上进行该实验。
Immunoprecipitation Image 1: Iba1/AIF-1 (E4O4W) XP® Rabbit mAb Expand Image
对 THP-1 细胞提取物中的 Iba1/AIF-1 进行免疫沉淀。泳道 1 为 10% input,泳道 2 为 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900,泳道 3 为 Iba1/AIF-1 (E4O4W) XP® Rabbit mAb。使用 Iba1/AIF-1 (E4O4W) XP® Rabbit mAb 进行蛋白质印迹分析。
Immunohistochemistry Image 1: Iba1/AIF-1 (E4O4W) XP® Rabbit mAb Expand Image
在 Leica® BOND Rx 上使用 Iba1/AIF-1 (E4O4W) XP® Rabbit mAb 对石蜡包埋的人神经内分泌癌细胞进行免疫组织化学分析。
Immunohistochemistry Image 2: Iba1/AIF-1 (E4O4W) XP® Rabbit mAb Expand Image
在 Leica® BOND Rx 上使用 Iba1/AIF-1 (E4O4W) XP® Rabbit mAb 对石蜡包埋的人非霍奇金淋巴瘤进行免疫组织化学分析。
Immunohistochemistry Image 3: Iba1/AIF-1 (E4O4W) XP® Rabbit mAb Expand Image
在 Leica® BOND Rx 上使用 Iba1/AIF-1 (E4O4W) XP® Rabbit mAb(左图)或 Iba1/AIF-1 (E5N4J) Mouse mAb (IHC Formulated) #58970(右图)对石蜡包埋的人卵巢透明细胞癌组织进行免疫组织化学分析。这两种抗体可检测人 Iba1/AIF-1 上的独立、独特表位。使用两种抗体获得的相似染色模式有助于确认染色特异性。
Immunohistochemistry Image 1: Iba1/AIF-1 (E4O4W) XP® Rabbit mAb Expand Image
使用 Iba1/AIF-1 (E4O4W) XP® Rabbit mAb(绿色)和 APP/β-Amyloid (NAB228) Mouse mAb #2450(红色)对石蜡包埋的人阿尔茨海默病的大脑进行双重免疫组织化学分析。
Immunohistochemistry Image 2: Iba1/AIF-1 (E4O4W) XP® Rabbit mAb Expand Image
使用 Iba1/AIF-1 (E4O4W) XP® Rabbit mAb 对石蜡包埋的小鼠脑进行免疫组织化学分析。
Immunohistochemistry Image 3: Iba1/AIF-1 (E4O4W) XP® Rabbit mAb Expand Image
使用 Iba1/AIF-1 (E4O4W) XP® Rabbit mAb 对石蜡包埋的小鼠脾脏进行免疫组织化学分析。
Immunohistochemistry Image 4: Iba1/AIF-1 (E4O4W) XP® Rabbit mAb Expand Image
使用 Iba1/AIF-1 (E4O4W) XP® Rabbit mAb 对石蜡包埋的小鼠小肠进行免疫组织化学分析。
Immunohistochemistry Image 5: Iba1/AIF-1 (E4O4W) XP® Rabbit mAb Expand Image
使用 Iba1/AIF-1 (E4O4W) XP® Rabbit mAb 对石蜡包埋的正常大鼠脑进行免疫组织化学分析。
Immunohistochemistry Image 6: Iba1/AIF-1 (E4O4W) XP® Rabbit mAb Expand Image
使用 Iba1/AIF-1 (E4O4W) XP® Rabbit mAb 对石蜡包埋的正常恒河猴脾脏进行免疫组织化学分析。
Immunohistochemistry Image 7: Iba1/AIF-1 (E4O4W) XP® Rabbit mAb Expand Image
使用 Iba1/AIF-1 (E4O4W) XP® Rabbit 对石蜡包埋的正常恒河猴肝脏进行免疫组织化学分析
Immunohistochemistry Image 8: Iba1/AIF-1 (E4O4W) XP® Rabbit mAb Expand Image
使用 Iba1/AIF-1 (E4O4W) XP® Rabbit mAb 对石蜡包埋的正常叙利亚仓鼠小肠进行免疫组织化学分析。
Immunohistochemistry Image 9: Iba1/AIF-1 (E4O4W) XP® Rabbit mAb Expand Image
使用 Iba1/AIF-1 (E4O4W) XP® Rabbit mAb对石蜡包埋的人卵巢浆液性癌进行免疫组织化学分析。
Immunohistochemistry Image 10: Iba1/AIF-1 (E4O4W) XP® Rabbit mAb Expand Image
使用 Iba1/AIF-1 (E4O4W) XP® Rabbit mAb 对石蜡包埋的人结肠癌进行免疫组织化学分析。
Immunohistochemistry Image 11: Iba1/AIF-1 (E4O4W) XP® Rabbit mAb Expand Image
以浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(右图)作为对照,使用 Iba1/AIF-1 (E4O4W) XP® Rabbit mAb(左图)对石蜡包埋的人乳腺导管癌细胞进行免疫组织化学分析。
Immunohistochemistry Image 12: Iba1/AIF-1 (E4O4W) XP® Rabbit mAb Expand Image
使用 Iba1/AIF-1 (E4O4W) XP® Rabbit mAb 对石蜡包埋的 THP-1 cell pellet 细胞沉淀物(左图,阳性)或 SH-SY5Y 细胞沉淀物(右图,阴性)进行免疫组织化学分析。
Immunofluorescence Image 1: Iba1/AIF-1 (E4O4W) XP® Rabbit mAb Expand Image
使用 Iba1/AIF-1 (E4O4W) XP® Rabbit mAb(绿色)对人皮层(左图)和小鼠 CA1 海马(右图)进行共聚焦免疫荧光分析。在小鼠组织切片中,细胞核标有 DAPI(蓝色)。图片由 Simone Brioschi 博士和 Marco Marcona 博士(华盛顿大学)提供,并经许可使用。
Immunofluorescence Image 2: Iba1/AIF-1 (E4O4W) XP® Rabbit mAb Expand Image
使用 Iba1/AIF-1 (E4O4W) XP® Rabbit mAb(绿色)对小鼠小肠进行共聚焦免疫荧光分析。肌动蛋白微丝由 DyLight 554 Phalloidin #13054(红色)标记。切片用 ProLong® Gold Antifade Reagent with DAPI #8961(蓝色)封装。
Immunofluorescence Image 3: Iba1/AIF-1 (E4O4W) XP® Rabbit mAb Expand Image
使用 Iba1/AIF-1 (E4O4W) XP® Rabbit mAb(绿色)对小鼠肝脏进行共聚焦免疫荧光分析。肌动蛋白微丝由 DyLight 554 Phalloidin #13054(红色)标记。切片用 ProLong® Gold Antifade Reagent with DAPI #8961(蓝色)封装。
Immunofluorescence Image 4: Iba1/AIF-1 (E4O4W) XP® Rabbit mAb Expand Image
使用 Iba1/AIF-1 (E4O4W) XP® Rabbit mAb(绿色)对小鼠海马进行共聚焦免疫荧光分析。切片用 ProLong® Gold Antifade Reagent with DAPI #8961(蓝色)封装。
Immunofluorescence Image 1: Iba1/AIF-1 (E4O4W) XP® Rabbit mAb Expand Image
使用 Iba1/AIF-1 (E4O4W) XP® Rabbit mAb(绿色)对用 TPA (12-O-Tetradecanoylphorbol-13-Acetate)(80 nM,24 小时;左图,阳性)分化的 THP-1 细胞和 SH-SY5Y 细胞(右图,阴性)进行共聚焦免疫荧光分析。肌动蛋白微丝由 DyLight 554 Phalloidin #13054(红色)标记。样本用 ProLong® Gold Antifade Reagent with DAPI #8961(蓝色)封片。
Flow Cytometry Image 1: Iba1/AIF-1 (E4O4W) XP® Rabbit mAb Expand Image
使用 Iba1/AIF-1 (E4O4W) XP® Rabbit mAb(实线)或浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(虚线)对 SH-SY5Y 细胞(蓝色,阴性)和 THP-1 细胞(绿色,阳性)进行流式细胞分析。Anti-rabbit IgG (H+L)、F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 作为二抗。