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97148
Phospho-Histone H2A.X (Ser139) (D7T2V) Mouse mAb (BSA and Azide Free)
Primary Antibodies
Monoclonal Antibody

Phospho-Histone H2A.X (Ser139) (D7T2V) Mouse mAb (BSA and Azide Free) #97148

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  1. WB
  2. IHC
  3. IF
  4. F
Western Blotting Image 1: Phospho-Histone H2A.X (Ser139) (D7T2V) Mouse mAb (BSA and Azide Free)
Western blot analysis of extracts from HeLa cells, untreated (-), UV-treated (+), or UV-treated and subsequently treated with calf intestinal phosphatase (CIP) plus λ-phosphatase (+), using Phospho-Histone H2A.X (Ser139) (D7T2V) Mouse mAb (upper) or Histone H2A (D6O3A) Rabbit mAb #12349 (lower). Data were generated using the standard formulation of this product.
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Immunohistochemistry Image 1: Phospho-Histone H2A.X (Ser139) (D7T2V) Mouse mAb (BSA and Azide Free)
Immunohistochemical analysis of paraffin-embedded human non-small cell lung carcinoma using Phospho-Histone H2A.X (Ser139) (D7T2V) Mouse mAb performed on the Leica® BOND Rx. Data were generated using the standard formulation of this product.
Immunohistochemistry Image 2: Phospho-Histone H2A.X (Ser139) (D7T2V) Mouse mAb (BSA and Azide Free)
Immunohistochemical analysis of paraffin-embedded human ductal breast carcinoma using Phospho-Histone H2A.X (Ser139) (D7T2V) Mouse mAb performed on the Leica® BOND Rx. Data were generated using the standard formulation of this product.
Immunohistochemistry Image 1: Phospho-Histone H2A.X (Ser139) (D7T2V) Mouse mAb (BSA and Azide Free)
Immunohistochemical analysis of paraffin-embedded human appendix using Phospho-Histone H2A.X (Ser139) (D7T2V) Mouse mAb. Data were generated using the standard formulation of this product.
Immunohistochemistry Image 2: Phospho-Histone H2A.X (Ser139) (D7T2V) Mouse mAb (BSA and Azide Free)
Immunohistochemical analysis of paraffin-embedded human breast carcinoma, untreated (left) or λ-phosphatase-treated (right), using Phospho-Histone H2A.X (Ser139) (D7T2V) Mouse mAb. Data were generated using the standard formulation of this product.
Immunohistochemistry Image 3: Phospho-Histone H2A.X (Ser139) (D7T2V) Mouse mAb (BSA and Azide Free)
Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using Phospho-Histone H2A.X (Ser139) (D7T2V) Mouse mAb. Data were generated using the standard formulation of this product.
Immunohistochemistry Image 4: Phospho-Histone H2A.X (Ser139) (D7T2V) Mouse mAb (BSA and Azide Free)
Immunohistochemical analysis of paraffin-embedded human lymph node using Phospho-Histone H2A.X (Ser139) (D7T2V) Mouse mAb. Data were generated using the standard formulation of this product.
Immunohistochemistry Image 5: Phospho-Histone H2A.X (Ser139) (D7T2V) Mouse mAb (BSA and Azide Free)
Immunohistochemical analysis of paraffin-embedded OVCAR-8 cell pellets, untreated (left) or UV-treated (right), using Phospho-Histone H2A.X (Ser139) (D7T2V) Mouse mAb. Data were generated using the standard formulation of this product.
Immunofluorescence Image 1: Phospho-Histone H2A.X (Ser139) (D7T2V) Mouse mAb (BSA and Azide Free)
Confocal immunofluorescent analysis of HeLa cells, untreated (left), treated with UV (100 mJ/cm2, 2 hr recovery; center), or treated with UV and post-processed with λ-phosphatase (right), using Phospho-Histone H2A.X (Ser139) (D7T2V) Mouse mAb (green). Actin filaments were labeled with DyLight 554 Phalloidin #13054 (red). Data were generated using the standard formulation of this product.
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Flow Cytometry Image 1: Phospho-Histone H2A.X (Ser139) (D7T2V) Mouse mAb (BSA and Azide Free)
Flow cytometric analysis of HeLa cells, untreated (blue) or treated with UV (100 mJ, 2 hr recovery; green), using Phospho-Histone H2A.X (Ser139) (D7T2V) Mouse mAb (solid lines) or concentration-matched Mouse (G3A1) mAb IgG1 Isotype Control #5415 (dashed lines). Anti-mouse IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4408 was used as a secondary antibody. Data were generated using the standard formulation of this product.
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To Purchase # 97148
Cat. # Size Qty. Price
97148SF
100 µg
$ 776

Supporting Data

REACTIVITY H M R Mk
SENSITIVITY Endogenous
MW (kDa) 15
Source/Isotype Mouse IgG1

Application Key:

  • WB-Western Blot
  • IP-Immunoprecipitation
  • IHC-Immunohistochemistry
  • ChIP-Chromatin Immunoprecipitation
  • C&R-CUT&RUN
  • C&T-CUT&Tag
  • DB-Dot Blot
  • eCLIP-eCLIP
  • IF-Immunofluorescence
  • F-Flow Cytometry

Species 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
  • All-All Species Expected

Product Usage Information

This product is the carrier-free version of product #80312. All data were generated using the same antibody clone in the standard formulation which contains BSA and glycerol.
This formulation is ideal for use with technologies requiring specialized or custom antibody labeling, including fluorophores, metals, lanthanides, and oligonucleotides. It is not recommended for ChIP, ChIP-seq, CUT&RUN, or CUT&Tag assays. If you require a carrier-free formulation for chromatin profiling, please contact us. Optimal dilutions/concentrations should be determined by the end user.

Formulation

Supplied in 1X PBS, BSA and Azide Free.

For standard formulation of this product see product #80312.

Storage

Store at -20°C. This product will freeze at -20°C so it is recommended to aliquot into single-use vials to avoid multiple freeze/thaw cycles. A slight precipitate may be present and can be dissolved by gently vortexing. This will not interfere with antibody performance.

Specificity / Sensitivity

Phospho-Histone H2A.X (Ser139) (D7T2V) Mouse mAb (BSA and Azide Free) recognizes endogenous levels of Histone H2A.X protein only when phosphorylated at Ser139.

Species Reactivity:

Human, Mouse, Rat, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser139 of human Histone H2A.X protein.

Background

Histone H2A.X is a variant histone that represents approximately 10% of the total H2A histone proteins in normal human fibroblasts (1). H2A.X is required for checkpoint-mediated cell cycle arrest and DNA repair following double-stranded DNA breaks (1). DNA damage, caused by ionizing radiation, UV-light, or radiomimetic agents, results in rapid phosphorylation of H2A.X at Ser139 by PI3K-like kinases, including ATM, ATR, and DNA-PK (2,3). Within minutes following DNA damage, H2A.X is phosphorylated at Ser139 at sites of DNA damage (4). This very early event in the DNA-damage response is required for recruitment of a multitude of DNA-damage response proteins, including MDC1, NBS1, RAD50, MRE11, 53BP1, and BRCA1 (1). In addition to its role in DNA-damage repair, H2A.X is required for DNA fragmentation during apoptosis and is phosphorylated by various kinases in response to apoptotic signals. H2A.X is phosphorylated at Ser139 by DNA-PK in response to cell death receptor activation, c-Jun N-terminal Kinase (JNK1) in response to UV-A irradiation, and p38 MAPK in response to serum starvation (5-8). H2A.X is constitutively phosphorylated on Tyr142 in undamaged cells by WSTF (Williams-Beuren syndrome transcription factor) (9,10). Upon DNA damage, and concurrent with phosphorylation of Ser139, Tyr142 is dephosphorylated at sites of DNA damage by recruited EYA1 and EYA3 phosphatases (9). While phosphorylation at Ser139 facilitates the recruitment of DNA repair proteins and apoptotic proteins to sites of DNA damage, phosphorylation at Tyr142 appears to determine which set of proteins are recruited. Phosphorylation of H2A.X at Tyr142 inhibits the recruitment of DNA repair proteins and promotes binding of pro-apoptotic factors such as JNK1 (9). Mouse embryonic fibroblasts expressing only mutant H2A.X Y142F, which favors recruitment of DNA repair proteins over apoptotic proteins, show a reduced apoptotic response to ionizing radiation (9). Thus, it appears that the balance of H2A.X Tyr142 phosphorylation and dephosphorylation provides a switch mechanism to determine cell fate after DNA damage.
  1. Yuan, J. et al. (2010) FEBS Lett 584, 3717-24.
  2. Rogakou, E.P. et al. (1998) J Biol Chem 273, 5858-68.
  3. Burma, S. et al. (2001) J Biol Chem 276, 42462-7.
  4. Rogakou, E.P. et al. (1999) J Cell Biol 146, 905-16.
  5. Mukherjee, B. et al. (2006) DNA Repair (Amst) 5, 575-90.
  6. Solier, S. et al. (2009) Mol Cell Biol 29, 68-82.
  7. Lu, C. et al. (2006) Mol Cell 23, 121-32.
  8. Lu, C. et al. (2008) FEBS Lett 582, 2703-8.
  9. Cook, P.J. et al. (2009) Nature 458, 591-6.
  10. Xiao, A. et al. (2009) Nature 457, 57-62.

Pathways & Proteins

Explore pathways + proteins related to this product.

Limited Uses

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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.

For Research Use Only. Not For Use In Diagnostic Procedures.
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