Cell Signaling Technology

Product Pathways - Neuroscience

EGR1 (44D5) Rabbit mAb #4154

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP IF-IC F ChIP H M R (B) Endogenous 75 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry  ChIP=Chromatin IP
Reactivity Key:  H=Human  M=Mouse  R=Rat  B=Bovine
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

EGR1 (44D5) Rabbit mAb detects endogenous levels of total EGR1 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the amino terminus of the sequence of human EGR1.

Western Blotting

Western Blotting

Western analysis of cell extracts from serum starved PC12, untreated or NGF-treated (100 ng/ml for 2 hours), using EGR1 (44D5) Rabbit mAb.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of PC-12 cells, untreated (blue) or NGF treated (green), using EGR1 (44D5) Rabbit mAb compared to a nonspecific negative control (red).

IF-IC

IF-IC

Confocal immunofluorescent analysis of dissociated PC12 cells either untreated (left) or NGF-treated for 2 hours (right), using EGR1 (44D5) Rabbit mAb (green) and MEK1/2 (L38C12) Mouse mAb (#4694) (red).


Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 THP-1 cells treated with TPA (12-O-Tetradecanoylphorbol-13-Acetate) #4174 (30ng/ml) overnight and either 10 μl of EGR1 (44D5) Rabbit mAb or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using human ATAD2 promoter primers, SimpleChIP® Human EGR1 Promoter Primers #5549, human PCM1 promoter primers, 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.

Background

EGR family members are transcriptional factors that contain three repetitive zinc finger DNA binding domains which bind to EGR response elements (ER) to regulate target gene expression (1). The expression of EGR family members is induced by growth factors, with EGR1 expression being induced by NGF (1,2). Increased EGR1 expression activates transcription of other signaling molecules, including CDK5 and tyrosine hydroxylase, and exerts long term effects on neural cell growth and differentiation (2,3).

  1. O'Donovan, K.J. et al. (1999) Trends Neurosci. 22, 167-173.
  2. Harada, T. et al. (2001) Nat. Cell Biol. 3, 453-459.
  3. Papanikolaou, N.A. and Sabban, E.L. (2000) J. Biol. Chem. 275, 26683-26689.

Application References

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Companion Products

Rabbit Monoclonals Produced Using Epitomics® Technology, U.S. Patent No. 5,675,063.

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

Products