Revision 6

#26768Store at -20C

1 Kit

(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.

    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 6
    #26768

    Parthanatos Antibody Sampler Kit

    Parthanatos Antibody Sampler Kit: Image 1 Expand Image
    Confocal immunofluorescent analysis of HeLa cells either untreated (left), treated with hydrogen peroxide (500 mM, 5 min; center), or treated with PARP inhibitor Talazoparib (100 nmol/L, 3 hr) followed by hydrogen peroxide (500 mM, 5 min; right) using Poly/Mono-ADP Ribose (E6F6A) Rabbit mAb #83732 (green), S6 Ribosomal Protein (54D2) Mouse mAb #2317 (red), and DAPI #4083 (blue).
    Parthanatos Antibody Sampler Kit: Image 2 Expand Image
    Simple Western™ analysis of lysates (1 mg/mL) from Jurkat cells treated with Etoposide (25 uM, 5 hours) using PARP (46D11) Rabbit mAb #9532. The virtual lane view (left) shows the target bands (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.
    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. Data is kindly provided by the laboratory of Dr. Gene Yeo and used with permission.
    Parthanatos Antibody Sampler Kit: Image 4 Expand Image
    Western blot analysis of extracts from Jurkat, C2C12, and C6 cells using AIF (D39D2) XP® Rabbit mAb.
    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
    After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
    Parthanatos Antibody Sampler Kit: Image 7 Expand Image
    Western blot analysis of extracts from various cell lines using MIF (E8S8H) Rabbit mAb (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). The absence of MIF protein in SNB-19 cells is consistent with mRNA expression profiles reported in public bioinformatic databases, confirming specificity of the antibody for MIF.
    Parthanatos Antibody Sampler Kit: Image 8 Expand Image
    Western blot analysis of Colo 205 cells treated (+) with combinations of the following treatments as indicated: hydrogen peroxide (500 μM, 5 min), hydrogen peroxide-treated lysates treated with phosphodiesterase 1 (0.5 μg/mL, 4 hr at 37ºC), or with tcPARG (5 μM, 4 hr at 37ºC ) using Poly/Mono-ADP Ribose (E6F6A) Rabbit mAb (upper), or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
    Parthanatos Antibody Sampler Kit: Image 9 Expand Image
    Western blot analysis of extracts from control HEK293 cells (lane 1) or PARP knockout HEK293 cells (lane 2) using PARP (46D11) Rabbit mAb #9532 (upper) or β-actin (13E5) Rabbit mAb #4970 (lower). The absence of signal in the PARP knockout HEK293 cells confirms the specificity of the antibody for PARP.
    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
    Confocal immunofluorescent analysis of mouse testis (left) and skeletal muscle (right) using MIF (E8S8H) 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).
    Parthanatos Antibody Sampler Kit: Image 12 Expand Image
    Western blot analysis of Colo 205 cells untreated (-), or treated with hydrogen peroxide (500 μM, 5 min; +), using Poly/Mono-ADP Ribose (E6F6A) Rabbit mAb (upper), or β-Tubulin (9F3) Rabbit mAb #2128 (lower).
    Parthanatos Antibody Sampler Kit: Image 13 Expand Image
    Western blot analysis of extracts from THP-1 cells, untreated or treated with TNF-α and cycloheximide as well as control extracts from SW620 and A20 cell lines, using PARP (46D11) Rabbit mAb.
    Parthanatos Antibody Sampler Kit: Image 14 Expand Image
    Confocal immunofluorescent analysis of mouse brainstem using AIF (D39D2) XP® Rabbit mAb #5318 (green) and GFAP (GA5) Mouse mAb #3670 (red). Sections were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
    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
    Confocal immunofluorescent analysis of U-87 MG cells (left, positive) or SNB-19 cells (right, negative) using MIF (E8S8H) 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).
    Parthanatos Antibody Sampler Kit: Image 17 Expand Image
    Confocal immunofluorescent analysis of mouse cerebellum using AIF (D39D2) XP® Rabbit mAb #5318 (green), GFAP (GA5) Mouse mAb #3670 (red), and F4/80 (BM8.1) Rat mAb #71299 (blue).
    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
    Confocal immunofluorescent analysis of mouse striatum using AIF (D39D2) XP® Rabbit mAb #5318 (green), GFAP (GA5) Mouse mAb #3670 (red), and F4/80 (BM8.1) Rat mAb #71299 (blue).
    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
    Confocal immunofluorescent analysis of mouse thalamus using AIF (D39D2) XP® Rabbit mAb #5318 (green), GFAP (GA5) Mouse mAb #3670 (red). Sections were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
    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
    Confocal immunofluorescent analysis of C2C12 cells using AIF (D39D2) XP® Rabbit mAb #5318 (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    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
    Confocal immunofluorescent analysis of HeLa cells using AIF (D39D2) XP Rabbit mAb (green), showing colocalization with mitochondria that have been labeled with MitoTracker® Red CMXRos (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    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).