Revision 7

#4154Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP, IF-IC, FC-FP, ChIP, ChIP-seq

REACTIVITY:

H M R

SENSITIVITY:

Endogenous

MW (kDa):

75

Source/Isotype:

Rabbit IgG

UniProt ID:

#P18146

Entrez-Gene Id:

1958

Product Information

Product Usage Information

For optimal ChIP and ChIP-seq results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
Immunofluorescence (Immunocytochemistry) 1:1600 - 1:6400
Flow Cytometry (Fixed/Permeabilized) 1:400 - 1:1600
Chromatin IP 1:50
Chromatin IP-seq 1:50

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

For a carrier free (BSA and azide free) version of this product see product #54070.

Specificity / Sensitivity

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

Species Reactivity:

Human, Mouse, Rat

Species predicted to react based on 100% sequence homology

Bovine

Source / Purification

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

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.

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

WB: Western Blotting IP: Immunoprecipitation IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized) ChIP: Chromatin IP ChIP-seq: Chromatin IP-seq

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 All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

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.

Revision 7
#4154

EGR1 (44D5) Rabbit mAb

Western Blotting Image 1: EGR1 (44D5) Rabbit mAb Expand Image
Western analysis of cell extracts from serum starved PC12, untreated or NGF-treated (100 ng/ml for 2 hours), using EGR1 (44D5) Rabbit mAb.
No image available
Immunofluorescence Image 1: EGR1 (44D5) Rabbit mAb Expand Image
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).
Flow Cytometry Image 1: EGR1 (44D5) Rabbit mAb Expand Image
Flow cytometric analysis of Jurkat cells, untreated (blue) or treated with TPA #4174 and Ionomycin, Calcium Salt #9995 (200 nM and 3 µM, 2 hr; green), using EGR1 (44D5) Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
Chromatin Immunoprecipitation Image 1: EGR1 (44D5) Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from THP-1 cells treated with TPA (12-O-Tetradecanoylphorbol-13-Acetate) #4174 (30ng/ml) overnight and EGR1 (44D5) Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across PCM1, a known target gene of EGR1 (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.
Chromatin Immunoprecipitation Image 2: EGR1 (44D5) Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from THP-1 cells treated with TPA (12-O-Tetradecanoylphorbol-13-Acetate) #4174 (30ng/ml) overnight and EGR1 (44D5) Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across chromosome 8 (upper), including PCM1 (lower), a known target gene of EGR1 (see additional figure containing ChIP-qPCR data).
Chromatin Immunoprecipitation Image 3: EGR1 (44D5) Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from THP-1 cells treated with TPA (12-O-Tetradecanoylphorbol-13-Acetate) #4174 (30ng/ml) overnight and either EGR1 (44D5) Rabbit mAb or 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.