Revision 9

#4154

Store at -20C

EGR1 (44D5) Rabbit mAb
CST Logo
Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

Web:

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Applications:
W, 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 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 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).

Background References

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 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

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.

限制使用

除非 CST 的合法授书代表以书面形式书行明确同意，否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件，除非书 CST 的合法授书代表以书面形式书独接受， 否书均被拒书，并且无效。

专品专有“专供研究使用”的专专或专似的专专声明， 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的， 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专，且专用于研专用途。将专品用于专断、专防或治专目的， 或专专售（专独或作专专成）或其他商专目的而专专专品，均需要 CST 的专独专可。客专：(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品，或使用专品制造任何商专专品，(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专，或使用专品开专任何与 CST 的专品或服专专争的专品或服专， (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专，(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品 ， (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 9

#4154

EGR1 (44D5) Rabbit mAb

Cell Signaling Technology Logo
Western blot analysis of extracts from PC-12 cells, untreated (-) or treated with human ꞵ-NGF(hꞵ-NGF; 100ng/ml, 2hr); +) using EGR1 (44D5) Rabbit mAb (upper) or ꞵ-Actin (D6A8) Rabbit mAb #8457 (lower). EGR1 is induced by human ꞵ-NGF treatment as expected.
Western Blotting Image 1: EGR1 (44D5) Rabbit mAb
Western blot analysis of extracts from various cell lines and tissues using EGR1 (44D5) Rabbit mAb (upper) or ꞵ-Actin (D6A8) Rabbit mAb #8457 (lower).
Western Blotting Image 2: EGR1 (44D5) Rabbit mAb

Immunoprecipitation of EGR1 from JHH-1 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 and lane 3 is EGR1 (44D5) Rabbit mAb. Western blot analysis was performed using EGR1 (44D5) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as a secondary antibody.
Immunoprecipitation Image 1: EGR1 (44D5) Rabbit mAb
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 9

#4154

EGR1 (44D5) Rabbit mAb

Cell Signaling Technology Logo
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).
Immunofluorescence Image 1: EGR1 (44D5) Rabbit mAb
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.
Flow Cytometry Image 1: EGR1 (44D5) Rabbit mAb
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 1: EGR1 (44D5) Rabbit mAb
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 9

#4154

EGR1 (44D5) Rabbit mAb

Cell Signaling Technology Logo
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 2: EGR1 (44D5) Rabbit mAb
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.
Chromatin Immunoprecipitation Image 3: EGR1 (44D5) Rabbit mAb
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.