Anti-Rad18 (mouse) antibody, rabbit polyclonal70-025100 µgShipping and Storage: Ship at 4℃ or -20℃ and store at -20℃
Form: 1 mg/ml IgG fraction in PBS- with 50% glycerol, filter-sterilized. Azide andcarrier-protein free
Reactivity: Mouse Rad18 protein. Not reactive to human Rad18.
Immunogengen: GST-fusion protein containing 100 carboxyl terminal amino acids ofmouse Rad18
Applications (see Ref 1~3)
1) Western blotting (1,000 fold dilution).
2) Immuno-precipitation (200~500 dilution))3) Indirect immuno-fluorescence staining. (assay dependent)
4) Immuno-histochemistry (100~300 fold dilution)
Background: The Rad6 (UBE2B)-Rad18 pair of genes plays a critical role inpost-replication repair of damaged DNA. Rad6 protein functions as an E2 enzyme andRad18 (509 aa, 57.4 kDa) as a ubiquitine ligase (E3) which ubiquitinates PCNA. Rad18
recruits translesion DNA polymerases to damaged DNA.
Database Link: UniProtKB/Swiss-Prot Q9QXK2 Mouse Rad18
Gene ID 16098139 Mouse Rad18

Fig.1.Identification of mouse Rad18
protein in ES cells by Western blot with
anti-mRad18 antibody.
WT; Lysate of wild-type mouse ES cells

RAD18-/-;

LysateofRad18

doubleknock-out mouse ES cells
Protein levels of α-tubulin in the lysates
are shown as a control.
Three bands are absent in RAD18
knock-out cells.
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Fig.2. Identification of mouse Rad18
protein in NIH3T3 cells by western blot
with the antibody. Cell extract, 23 ug,was used. Anti-mouse Rad18 was used
at 2,000 fold dilution. Similar to Fig.1,
two extra bands (75~90 kDa) may
represent

modifiedproducts(ubiquitination, phosphorylation).
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Fig.3. Immunofluorescence staining of Rad18

Fig.4. Detection of Rad18 in mouse testis.

proteinantibody.

Section of paraformaldehyde fixed mouse was

Wild-type (+/+) and RAD18-/- ES cells (-/-).

stained with anti-mRad18 antibody. As a second

Samples were prefixed 3.7% formaldehyde

antibody, peroxidase-conjugated anti-rabbit IgG

and fixed with 80% methanol. Anti-mRad18

donky antibody was used. Signals were enhanced

antibody was used at 1/300 dilution. AS awith TSA plus biotin system and detected bysecond antibody,FITC conjugated anti-rabbit

using DAB substrate.

IgG was used

Arrows indicate undifferentiated spermatogonia.

withanti-mRad18

Related Products: 70-020 anti-Rad6 antibody

70-023 anti-Rad18 (human) antibody

References: This product has been used in the following publications.
1. Tateishi S. et al. (2003) Enhanced genomic instability and defective postreplication
repair in RAD18 knockout mouse embryonic stem cells. Mol Cell Biol 23:474-81.
PubMed 12509447 WB, IF/IC
2. Watanabe K. et al. (2004) Rad18 guides poleta to replication stalling sites throughphysical interaction and PCNA monoubiquitination. EMBO J. 23:3886-96. PubMed
15359278 WB
3. Masuyama S. et al. (2005) Regulated expression and dynamic changes in subnuclear
localization of mammalian Rad18 under normal and genotoxic conditions. Genes
Cells. 10:753-62. PubMed 16098139 IHC
4. Sun J. et al. (2009) Rad18 is required for long-term maintenance of spermatogenesis
in mouse testes. Mech Dev 126:173-83.PubMed 19068231 IHC, WB
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Tel. 81-6-6877-2335

Fax. 81-6-6877-2336

info@bioacademia.co.jp

http://www.bioacademia.co.jp/en/