Egr1 Rabbit Polyclonal Antibody
cat.: ER1907-98
Product Type: Rabbit polyclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IHC-P, IF-Cell, IF-Tissue, FC
Clonality: Polyclonal
Form: Liquid
Storage condition: Store at -20℃ for one year. Avoid repeated freeze/thaw cycles. The lyophilized antibody is stable at room temperature for at least one month and for greater than a year when kept at -20℃. When reconstituted in sterile pH 7.4 0.01M PBS or diluent of antibody the antibody is stable for at least two weeks at 2-4℃.
Storage buffer: 0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: 60 KDa
Isotype: IgG
Immunogen: KLH conjugated synthetic peptide derived from human Egr-1 401-500/453
Positive control: Mouse small intestine, U-937 cells, Mouse Lung Lysate, rat small intestine tissue, rat liver tissue.
Subcellular location: Nucleus.
Recommended Dilutions:
  WB
  IHC-P
  IF-cell
  IF-tissue
  FC
1:500-2000
1:100-500
1:100
1:100-500
2ug/test
Uniprot #: SwissProt: P18146 Human
Alternative names: AT225 Early growth response 1 Early growth response protein 1 EGR 1 EGR-1 EGR1 EGR1_HUMAN G0S30 KROX 24 KROX24 Nerve growth factor-induced clone A Nerve growth factor-induced protein A NGFI-A NGFIA TIS8 Transcription factor ETR103 Transcription factor Zif268 ZIF 268 ZIF268 Zinc finger protein 225 Zinc finger protein Krox-24 ZNF225
Images
ER1907-98_1.jpg Fig1: Sample:
Lung (Mouse) Lysate at 40 ug
Primary: Anti-Egr1 (bs-1076R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 60 kD
Observed band size: 60 kD
ER1907-98_2.jpg Fig2: Sample:
Liver (Rat) Lysate at 40 ug
Primary: Anti- Egr1 (bs-1076R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 60 kD
Observed band size: 60 kD
ER1907-98_3.jpg Fig3: Tissue/cell: rat small intestine tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-Egr1 Polyclonal Antibody, Unconjugated(bs-1076R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
ER1907-98_4.jpg Fig4: Tissue/cell: rat liver tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-Egr1 Polyclonal Antibody, Unconjugated(bs-1076R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
ER1907-98_5.jpg Fig5: Paraformaldehyde-fixed, paraffin embedded (Mouse small intestine); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Egr1) Polyclonal Antibody, Unconjugated (bs-1076R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
ER1907-98_6.jpg Fig6: A431 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37°C for 20 min; Antibody incubation with (Egr1) polyclonal Antibody, Unconjugated (bs-1076R) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.
ER1907-98_7.jpg Fig7: Blank control:K562.
Primary Antibody (green line): Rabbit Anti-Egr1 antibody (bs-1076R)
Dilution: 2μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-FITC
Dilution: 0.5μg /test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 0.1% PBST for 20 min at room temperature. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
ER1907-98_8.jpg Fig8: U-937 cells were fixed with 4% PFA for 10min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at room temperature,and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with Egr1 Antibody(bs-1076R) at 1:500 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed.Cells stained with primary antibody (green), and isotype control (orange).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.