MDC1 antibody
Sheep anti Human MDC1 antibody (AHP799) used for the visualization of MDC1 expression in U2OS osteosarcoma cells following irradiation induced DNA damage by immunofluorescence.
Image caption:
PP4 dephosphorylates γH2AX at the sites of DNA damage. (A,B) U2OS cells transfected with siRNAs against PP4C, PP2ACα/β or a control sequence were irradiated with 1.5 Gy and processed for γH2AX (A,B) and MDC1 (B) immunofluorescence and DAPI counterstaining at the indicated time points. Images were taken with a × 40 objective (A) or with a × 63 objective (B). Scale bar, 23 μm (A) and 16 μm (B). (C–F) Quantification of cells with >5 γH2AX (C), >10 γH2AX (D,F) and >5 MDC1 (E) foci. At least 300 cells were analysed per experiment. Cells were treated with KU55933 and KU57788 12 min post-IR (F). Error bars represent standard error of mean of independent duplicated experiments (C) or standard deviation of three independent experiments (D–F). A × 63 oil immersion objective (C,E) or a × 40 water immersion objective (D,F) was used. (G) U2OS cells transfected with siRNAs against PP4C or a control sequence were collected at the indicated time points immediately before (no IR), or 1 or 6 h after irradiation (10 Gy), then fractionated as described in Méndez & Stillman (2000) into whole-cell lysates (WCLs), nuclear soluble (S3) or chromatin (P3) fractions, and analysed by immunoblotting using the indicated antibodies. DAPI, 4,6-diamidino-2-phenylindole; H3, histone H3; IR, irradiation; MDC1, mediator of DNA damage checkpoint 1; siRNA, small interfering RNA.
From: Nakada S, Chen GI, Gingras AC, Durocher D.
PP4 is a gamma H2AX phosphatase required for recovery from the DNA damage checkpoint.
EMBO Rep. 2008 Oct;9(10):1019-26.
Sheep anti Human MDC1 antibody (AHP799) used for the demonstration of MDC1 accumulation of MDC1 in murine spermatazoa by immunofluorescence.
Image caption:
Analysis of epigenetic differences in meiotic silencing in oocytes compared to spermatocytes. a XY pachytene spermatocyte stained for SYCP3 (magenta), HORMAD2 (green) and MDC1 (red), showing MDC1 accumulation in the sex chromatin (arrow). b XO pachytene oocyte stained for SYCP3 (magenta), HORMAD2 (green) and MDC1 (red), also showing MDC1 enrichment on the asynapsed X chromosome (arrow). c XY pachytene spermatocyte stained for SYCP3 (magenta), HORMAD2 (green) and SUMO-1 (red), showing accumulation of SUMO-1 in the sex chromatin (arrow). d XO pachytene oocyte stained for SYCP3 (magenta), HORMAD2 (green) and SUMO-1 (red), also showing SUMO-1 enrichment on the asynapsed X chromosome (arrow). e XY diplotene spermatocyte stained with SYCP3 to mark chromosome axes (magenta), HORMAD2 (green) to label the asynapsed X and Y chromosomes (arrow) and H3K9me3 (red), which shows enrichment in the chromatin of the X and Y chromosomes and also at constitutive heterochromatin (asterisks). A line (yellow) was drawn through the asynapsed X (arrow) to quantify HORMAD2 and H3K9me3 intensities (g). f XO diplotene oocyte showing no enrichment of H3K9me3 in the chromatin of the asynapsed X chromosome (arrow). H3K9me3 staining is restricted to sites of constitutive heterochromatin (asterisks). A line (yellow) was drawn through the asynapsed X (arrow) to quantify HORMAD2 and H3K9me3 intensities (h). g, h Intensities of H3K9me3 and HORMAD2 immunostaining quantified by densitometry across the indicated paths along (g) the asynapsed X in the spermatocyte from panel e, and along (h) the asynapsed X in diplotene XO oocyte from panel f. Scale bars = 10 μm
From: Cloutier JM, Mahadevaiah SK, ElInati E, Tóth A, Turner J.
Mammalian meiotic silencing exhibits sexually dimorphic features.
Chromosoma. 2016 Jun;125(2):215-26.
Sheep anti Human MDC1 antibody recognizes human Mediator of DNA damage checkpoint protein 1, also known as MDC1 or Nuclear factor with BRCT domains 1. MDC1 is a 2089 amino acid, ~250 kDa nuclear protein containing two BRCT domains and a single FHA domain. MDC1 is involved in checkpoint mediated cell cycle arest in response to DNA damage (UniProt: Q14676)
Product Details
- Target Species
- Human
- Product Form
- Serum - liquid
- Antiserum Preparation
- Antisera to human MDC1 (3838) were raised by repeated immunisations of sheep with highly purified antigen.
- Preservative Stabilisers
0.09% Sodium Azide - Immunogen
- Recombinant MDC1 BRCT domains (aa 1883 - 2089).
Storage Information
- Storage
- Store at +4oC or at -20oC if preferred.
This product should be stored undiluted.
Storage in frost-free freezers is not recommended. - Shelf Life
- 18 months from date of despatch.
More Information
- UniProt
- Q14676 Related reagents
- Entrez Gene
- MDC1 Related reagents
- Regulatory
- For research purposes only
Applications of MDC1 antibody
| Application Name | Verified | Min Dilution | Max Dilution |
|---|---|---|---|
| Immunofluorescence | 1/500 | ||
| Western Blotting | 1/2000 |
Where this antibody has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the antibody for use in their own systems using appropriate negative/positive controls.
- Western Blotting
- AHP799 detects 3 bands of approximately 250 kDa in HeLa and 293T cell lysates.
Copyright © 2018 Bio-Rad Antibodies (formerly AbD Serotec)
Secondary Antibodies Available
| Description | Product Code | Pack Size | Applications | List Price | Quantity |
|---|---|---|---|---|---|
| Rabbit anti Sheep IgG (H/L):Biotin | 5184-2304 | 1.5 ml | C E F WB | ||
| Donkey anti Sheep/Goat IgG:DyLight®488 | STAR88D488GA | 0.1 mg | F IF | ||
| Donkey anti Sheep/Goat IgG:DyLight®549 | STAR88D549GA | 0.1 mg | F IF | ||
| Donkey anti Sheep/Goat IgG:DyLight®649 | STAR88D649GA | 0.1 mg | F IF | ||
| Donkey anti Sheep/Goat IgG:FITC | STAR88F | 1 mg | C | ||
| Donkey anti Sheep/Goat IgG:HRP | STAR88P | 1 ml | C E P WB |
Product Specific References
References for MDC1 antibody
-
Goldberg, M. et al. (2003) MDC1 is required for the intra-S-phase DNA damage checkpoint.
Nature. 421 (6926): 952-6. -
Lou, Z. et al. (2003) MDC1 is coupled to activated CHK2 in mammalian DNA damage response pathways.
Nature. 421 (6926): 957-61. -
Shi, W. et al. (2008) Disassembly of MDC1 foci is controlled by ubiquitin-proteasome-dependent
degradation.
J Biol Chem. 283: 31608-16. -
Rai, R. et al (2008) Differential regulation of centrosome integrity by DNA damage response proteins.
Cell Cycle. 7: 2225-33. -
Ichijima, Y. et al. (2011) MDC1 directs chromosome-wide silencing of the sex chromosomes in male germ cells.
Genes Dev. 25: 959-71. -
Sin, H.S. et al. (2012) Human postmeiotic sex chromatin and its impact on sex chromosome evolution.
Genome Res. 22: 827-36. -
Nakada, S. et al. (2008) PP4 is a gamma H2AX phosphatase required for recovery from the DNA damage checkpoint.
EMBO Rep. 9: 1019-26. -
Nakada, S. et al. (2010) Non-canonical inhibition of DNA damage-dependent ubiquitination by OTUB1.
Nature. 466: 941-6. -
Reichert, S. et al. (2011) Survivin inhibition and DNA double-strand break repair: A molecular mechanism to overcome radioresistance in glioblastoma
Radiother Oncol. 101: 51-8. -
Capalbo, G. et al. (2010) Radiation-induced survivin nuclear accumulation is linked to DNA damage repair.
Int J Radiat Oncol Biol Phys. 77 (1): 226-34. -
Cloutier, J.M. et al. (2016) Mammalian meiotic silencing exhibits sexually dimorphic features.
Chromosoma. 125 (2): 215-26. -
Kato, Y. et al. (2015) FANCB is essential in the male germline and regulates H3K9 methylation on the sex chromosomes during meiosis.
Hum Mol Genet. 24 (18): 5234-49.
Further Reading
-
Stewart, G.S. et al. (2003) MDC1 is a mediator of the mammalian DNA damage checkpoint.
Nature. 421 (6926): 961-6.
