Revision 6

#26768Store at -20C

1 个试剂盒

(4 x 20 microliters)

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.
Product Includes Product # Quantity Mol. Wt Isotype/Source
PARP (46D11) Rabbit mAb 9532 20 µl 116, 89 kDa Rabbit 
AIF (D39D2) XP® Rabbit mAb 5318 20 µl 67 kDa Rabbit IgG
MIF (E8S8H) Rabbit mAb 75038 20 µl 12 kDa Rabbit IgG
Poly/Mono-ADP Ribose (E6F6A) Rabbit mAb 83732 20 µl Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

The Parthanatos Antibody Sampler Kit provides an economical means of detecting the activation of parthanatos. The kit includes enough antibodies to perform two western blot experiments with each primary antibody.

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 antibodies.

Background

Parthanatos is a form of regulated cell death that follows a multistep cascade and is triggered by the accumulation of poly (ADP-ribose) (PAR). When PAR polymerase-1 (PARP-1) is overactivated under certain conditions, excessive PAR is produced and binds to apoptosis-inducing factor (AIF). As a result, AIF is released from the mitochondria and forms a complex with macrophage migration inhibitory factor (MIF). Subsequently, the AIF/MIF complex is translocated to the nucleus where MIF cleaves genomic DNA into large fragments, and cell death follows (1-3). Studies have found that parthanatos is involved in the pathogenesis of many diseases, particularly neurodegenerative disorders, such as Alzheimer's disease, Huntington's disease, Parkinson's disease, and amyotrophic lateral sclerosis (ALS) (4-7).

  1. Fatokun, A.A. et al. (2014) Br J Pharmacol 171, 2000-16.
  2. Wang, Y. et al. (2016) Science 354, aad6872. doi: 10.1126/science.aad6872.
  3. Liu, L. et al. (2022) Cell Mol Life Sci 79, 60.
  4. Park, H. et al. (2020) Int Rev Cell Mol Biol 353, 1-29.
  5. Wang, X. and Ge, P. (2020) Neuroscience 449, 241-250.
  6. Zhou, Y. et al. (2021) Pharmacol Res 163, 105299.
  7. Thapa, K. et al. (2021) Life Sci 267, 118975.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    XP is a registered trademark of Cell Signaling Technology, Inc.
    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
    #26768

    Parthanatos 抗体小包装组合

    Parthanatos Antibody Sampler Kit: Image 1 Expand Image
    使用 Poly/Mono-ADP Ribose (E6F6A) Rabbit mAb #83732(绿色)、S6 Ribosomal Protein (54D2) Mouse mAb #2317(红色)和 DAPI #4083(蓝色)对未经处理(左图)、经过氧化氢处理(500 mM,5 分钟;中图)或经 PARP 抑制剂 Talazoparib(100 nmol/L,3 小时)处理后再经过氧化氢处理(500 mM,5 分钟;右图)的 HeLa 细胞进行共聚焦免疫荧光分析。
    Parthanatos Antibody Sampler Kit: Image 2 Expand Image
    使用 PARP (46D11) Rabbit mAb #9532 对经过 Etoposide(25 uM,5 小时)处理的 Jurkat 细胞的裂解物 (1 mg/mL) 进行 Simple Western™ 分析。虚拟泳道式图像(左图)显示一抗稀释比例在 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。
    Parthanatos Antibody Sampler Kit: Image 3 Expand Image
    Enhanced cross-linking and immunoprecipitation (eCLIP) was performed with RNA from K-562 cells and PARP (46D11) Rabbit mAb using a protocol based on the RBP-eCLIP Kit from EclipseBio. The figure shows binding across the GET4 transcript. 数据由 Gene Yeo 博士的实验室友情提供,并经许可使用。
    Parthanatos Antibody Sampler Kit: Image 4 Expand Image
    使用 AIF (D39D2) XP® Rabbit mAb,对 Jurkat、C2C12 和 C6 细胞提取物进行蛋白质印迹分析。
    Parthanatos Antibody Sampler Kit: Image 5 Expand Image
    Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using AIF (D39D2) XP® Rabbit mAb.
    Parthanatos Antibody Sampler Kit: Image 6 Expand Image
    一抗与靶标蛋白结合之后,与偶联 HRP 的二抗形成复合体。添加 LumiGLO®,在酶催化分解期间发光。
    Parthanatos Antibody Sampler Kit: Image 7 Expand Image
    使用 MIF (E8S8H) Rabbit mAb(上图)和 GAPDH (D16H11) XP® Rabbit mAb #5174(下图)对不同细胞系的提取物进行蛋白质印迹分析。SNB-19 细胞中 MIF 蛋白的缺失与公开的生物信息数据库中报道的 mRNA 表达谱一致,从而证实了该抗体对 MIF 的特异性。
    Parthanatos Antibody Sampler Kit: Image 8 Expand Image
    使用 Poly/Mono-ADP Ribose (E6F6A) Rabbit mAb(上图)或 GAPDH (D16H11) XP® Rabbit mAb #5174(下图)对未经处理 (-) 或已经过以下所示的联合处理方法处理的 Colo 205 细胞进行蛋白质印迹分析:用过氧化氢(500 μM,5 分钟)处理,或经过氧化氢处理的裂解物再用磷酸二酯酶 1(0.5 μg/mL,4 小时,在 37ºC 下)或 tcPARG(5 μM,4 小时,在 37ºC 下)处理。
    Parthanatos Antibody Sampler Kit: Image 9 Expand Image
    使用 PARP (46D11) Rabbit mAb #9532(上图)或 β-actin (13E5) Rabbit mAb #4970(下图)对对照 HEK293 细胞(泳道1)或 PARP 敲除型 HEK293 细胞(泳道2提取物进行蛋白质印迹法分析。PARP 敲除型 HEK293 细胞中没有信号,这证实了该抗体对 PARP 的特异性。
    Parthanatos Antibody Sampler Kit: Image 10 Expand Image
    Immunohistochemical analysis of paraffin-embedded human papillary thyroid carcinoma using AIF (D39D2) XP® Rabbit mAb.
    Parthanatos Antibody Sampler Kit: Image 11 Expand Image
    使用 MIF (E8S8H) Rabbit mAb (绿色)对小鼠结肠细胞(左图)和骨骼肌细胞(右图)进行共聚焦免疫荧光分析。肌动蛋白丝用 DyLight 554 Phalloidin #13054(红色)标记。样品用 ProLong® Gold Antifade Reagent with DAPI #8961(蓝色)封装。
    Parthanatos Antibody Sampler Kit: Image 12 Expand Image
    使用 Poly/Mono-ADP Ribose (E6F6A) Rabbit mAb(上图)或 β-Tubulin (9F3) Rabbit mAb #2128(下图)对未经 (-) 或已经过氧化氢(500μM,5 分钟;+)处理的 Colo 205 细胞进行蛋白质印迹分析。
    Parthanatos Antibody Sampler Kit: Image 13 Expand Image
    使用 PARP (46D11) Rabbit mAb 对未处理或经 TNF-α 和 cycloheximide 处理的 THP-1 细胞提取物、以及 SW620 和 A20 细胞系的对照提取物进行蛋白质印迹分析。
    Parthanatos Antibody Sampler Kit: Image 14 Expand Image
    使用 AIF (D39D2) XP® Rabbit mAb #5318(绿色)和 GFAP (GA5) Mouse mAb #3670(红色)对小鼠脑干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。
    Parthanatos Antibody Sampler Kit: Image 15 Expand Image
    Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma using AIF (D39D2) XP® Rabbit mAb.
    Parthanatos Antibody Sampler Kit: Image 16 Expand Image
    使用 MIF (E8S8H) Rabbit mAb(绿色)对 U-87 MG 细胞(左图,阳性)或 SNB-19 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。
    Parthanatos Antibody Sampler Kit: Image 17 Expand Image
    使用 AIF (D39D2) XP® Rabbit mAb #5318(绿色)、GFAP (GA5) Mouse mAb #3670(红色)和 F4/80 (BM8.1) Rat mAb #71299(蓝色)对小鼠小脑进行共聚焦免疫荧光分析。
    Parthanatos Antibody Sampler Kit: Image 18 Expand Image
    Immunohistochemical analysis of paraffin-embedded human renal cell carcinoma using AIF (D39D2) XP® Rabbit mAb.
    Parthanatos Antibody Sampler Kit: Image 19 Expand Image
    使用 AIF (D39D2) XP® Rabbit mAb #5318(绿色)、GFAP (GA5) Mouse mAb #3670(红色)和 F4/80 (BM8.1) Rat mAb #71299(蓝色)对小鼠纹状体进行共聚焦免疫荧光分析。
    Parthanatos Antibody Sampler Kit: Image 20 Expand Image
    Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using AIF (D39D2) XP® Rabbit mAb.
    Parthanatos Antibody Sampler Kit: Image 21 Expand Image
    使用 AIF (D39D2) XP® Rabbit mAb #5318(绿色)和 GFAP (GA5) Mouse mAb #3670(红色)对小鼠丘脑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。
    Parthanatos Antibody Sampler Kit: Image 22 Expand Image
    Immunohistochemical analysis of paraffin-embedded human papillary carcinoma of the breast using AIF (D39D2) XP® Rabbit mAb.
    Parthanatos Antibody Sampler Kit: Image 23 Expand Image
    使用 AIF (D39D2) XP® Rabbit mAb #5318(绿色),对 C2C12 细胞进行共聚焦免疫荧光分析。使用 DY-554 phalloidin(红色)标记肌动蛋白纤丝。蓝色伪彩 = DRAQ5® #4084(DNA 荧光染料)。
    Parthanatos Antibody Sampler Kit: Image 24 Expand Image
    Immunohistochemical analysis of paraffin-embedded normal human testis using AIF (D39D2) XP® Rabbit mAb.
    Parthanatos Antibody Sampler Kit: Image 25 Expand Image
    使用 AIF (D39D2) XP Rabbit mAb(绿色),对显示与用 MitoTracker® Red CMXRos(红色)标记的线粒体共定位的 Hela 细胞进行共聚焦免疫荧光分析。蓝色伪彩 = DRAQ5® #4084(DNA 荧光染料)。
    Parthanatos Antibody Sampler Kit: Image 26 Expand Image
    Immunohistochemical analysis of paraffin-embedded normal human spleen using AIF (D39D2) XP® Rabbit mAb.
    Parthanatos Antibody Sampler Kit: Image 27 Expand Image
    Immunohistochemical analysis of paraffin-embedded normal human skeletal muscle using AIF (D39D2) XP® Rabbit mAb.
    Parthanatos Antibody Sampler Kit: Image 28 Expand Image
    Immunohistochemical analysis of paraffin-embedded normal human liver using AIF (D39D2) XP® Rabbit mAb.
    Parthanatos Antibody Sampler Kit: Image 29 Expand Image
    Immunohistochemical analysis of paraffin-embedded normal human kidney using AIF (D39D2) XP® Rabbit mAb.
    Parthanatos Antibody Sampler Kit: Image 30 Expand Image
    Immunohistochemical analysis of paraffin-embedded normal human breast using AIF (D39D2) XP® Rabbit mAb.
    Parthanatos Antibody Sampler Kit: Image 31 Expand Image
    Immunohistochemical analysis of paraffin-embedded A20 syngeneic tumor using AIF (D39D2) XP® Rabbit mAb.
    Parthanatos Antibody Sampler Kit: Image 32 Expand Image
    Immunohistochemical analysis of paraffin-embedded LL/2 syngeneic tumor using AIF (D39D2) XP® Rabbit mAb.
    Parthanatos Antibody Sampler Kit: Image 33 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse testis using AIF (D39D2) XP® Rabbit mAb.
    Parthanatos Antibody Sampler Kit: Image 34 Expand Image
    Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using AIF (D39D2) XP® Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).