EGFR Rabbit Polyclonal Antibody

• Catalog Number: orb10585
• Category: Antibodies
• Description: EGFR Rabbit Polyclonal Antibody
• Target: EGFR
• Clonality: Polyclonal
• Species/Host: Rabbit
• Isotype: IgG
• Conjugation: Unconjugated
• Reactivity: Human
• Predicted Reactivity: Mouse, Rabbit, Rat
• Form/Appearance: Liquid
• Concentration: 1mg/ml
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Purification: Affinity purified by Protein A
• Immunogen: KLH conjugated synthetic peptide derived from human EGFR (801-900/1210aa)
• UniProt ID: P00533
• RRID: AB_10934660
• MW: 120 kDa
• Tested applications: FC, IF, IHC-Fr, IHC-P
• Dilution range: IHC-P=1:100-500, IHC-F=1:100-500, IF=1:100-500, Flow-Cyt=1μg/Test
• Antibody Type: Primary Antibody
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Alternative names: Avian erythroblastic leukemia viral(v erb b) oncog
• Research Area: Cancer Research, Cell Biology, Cell Differentiatio
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Blank control: HUVEC cells (blue). Primary Antibody: Rabbit Anti-EGFR antibody (orb10585), Dilution: 1 µg in 100 µl 1X PBS containing 0.5% BSA, Isotype Control Antibody: Rabbit IgG (orange), used under the same conditions, Secondary Antibody: Goat anti-rabbit IgG-PE (white blue), Dilution: 1:200 in 1 X PBS containing 0.5% BSA. Protocol, The cells were fixed with 2% paraformaldehyde (10 min), then permeabilized with 90% ice-cold methanol for 30 min on ice. Primary antibody (orb10585, 1 µg/1x10^6 cells) were incubated for 30 min on the ice, followed by 1 X PBS containing 0.5% BSA + 10% goat serum (15 min) to block non-specific protein-protein interactions. Then the Goat Anti-rabbit IgG/PE antibody was added into the blocking buffer mentioned above to react with the primary antibody at 1/200 dilution for 30 min on ice. Acquisition of 20000 events was performed.

Paraformaldehyde-fixed, paraffin embedded (Human brain glioma), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (EGFR) Polyclonal Antibody, Unconjugated (orb10585) at 1:100 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (orb868589) for 90 minutes, and DAPI for nuclei staining.

Paraformaldehyde-fixed, paraffin embedded (Human brain glioma), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (EGFR) Polyclonal Antibody, Unconjugated (orb10585) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (Human lung carcinoma), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (EGFR) Polyclonal Antibody, Unconjugated (orb10585) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.