Revision 1
#40291
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For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:
W, IP, ChIP
Reactivity:
H M R Mk
Sensitivity:
Endogenous
MW (kDa):
62
Source/Isotype:
Rabbit IgG
UniProt ID:
#Q16254
Entrez-Gene Id:
1874
Product Usage Information
Application | Dilution |
---|---|
Western Blotting | 1:1000 |
Immunoprecipitation | 1:50 |
Chromatin IP | 1:50 |
Storage
Specificity/Sensitivity
E2F4 (E3G2G) Rabbit mAb recognizes endogenous levels of total E2F4 protein.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu405 of human E2F4 protein.
Background
The E2F family consists of 8 transcription factors that regulate genes that control cell cycle progression by complexing with DP and Rb proteins (1-4). E2F transcriptional activation is generally opposed by associating with RB proteins, pRB, p107, and p130 (5-7). E2F-1, -2, and -3a function as activators that can help quiescent cells enter S phase, while E2F-3b, -4, and -5 repress cell growth through the recruitment of HDAC’s and other corepressors to target genes (8-10). E2F-6 diverges considerably from other family members, and has repressive properties governed not through interaction with Rb proteins, but by recruiting the polycomb repressive complex (11,12). E2F-7, and -8 are unique in that they have two DNA-binding domains and do not heterodimerize with DP proteins. These E2F family members repress transcription and delay progression of the cell cycle through the regulation of E2F-1 (13-15)
Background References
- Nevins, J.R. (1992) Nature 358, 375-6.
- Wu, C.L. et al. (1995) Mol Cell Biol 15, 2536-46.
- Huber, H.E. et al. (1993) Proc Natl Acad Sci U S A 90, 3525-9.
- Rogers, K.T. et al. (1996) Proc Natl Acad Sci U S A 93, 7594-9.
- Shirodkar, S. et al. (1992) Cell 68, 157-66.
- Hiebert, S.W. et al. (1992) Genes Dev 6, 177-85.
- Dyson, N. (1998) Genes Dev 12, 2245-62.
- DeGregori, J. et al. (1997) Proc Natl Acad Sci U S A 94, 7245-50.
- Lukas, J. et al. (1996) Mol Cell Biol 16, 1047-57.
- Meloni, A.R. et al. (1999) Proc Natl Acad Sci U S A 96, 9574-9.
- Trimarchi, J.M. et al. (1998) Proc Natl Acad Sci U S A 95, 2850-5.
- Trimarchi, J.M. et al. (2001) Proc Natl Acad Sci U S A 98, 1519-24.
- Logan, N. et al. (2004) Oncogene 23, 5138-50.
- Logan, N. et al. (2005) Oncogene 24, 5000-4.
- Moon, N.S. and Dyson, N. (2008) Dev Cell 14, 1-3.
Species Reactivity
Species reactivity is determined by testing in at least one approved application (e.g., western blot).
Western Blot Buffer
IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
Applications Key
W: Western Blotting IP: Immunoprecipitation ChIP: Chromatin IP
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 GP: Guinea Pig Rab: Rabbit G: Goat All: All Species Expected
Trademarks and Patents
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XP is a registered trademark of Cell Signaling Technology, Inc.
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Revision 1
#40291
E2F4 (E3G2G) Rabbit mAb
Western blot analysis of extracts from various cell lines using E2F4 (E3G2G) Rabbit mAb
Immunoprecipitation of E2F4 from HT-1080 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is E2F4 (E3G2G) Rabbit mAb. Western blot analysis was performed with E2F4 (E3G2G) Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used as a secondary antibody.
Chromatin immunoprecipitations were performed with cross-linked chromatin from Raji cells and either E2F4 (E3G2G) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human Timeless Intron 1 Primers #7001, SimpleChIP® Human DHFR Intron 1 Primers #7531 and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.