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Anti-EYA4 Antibody

Catalog Number: orb654424

DispatchUsually dispatched within 3-4 weeks
$ 210.00
Catalog Numberorb654424
CategoryAntibodies
DescriptionAnti-EYA4 Antibody. Tested in ELISA, Flow Cytometry, IHC, WB applications. This antibody reacts with Human, Monkey, Rat.
Species/HostRabbit
ClonalityPolyclonal
Tested applicationsELISA, FC, IHC, WB
ReactivityHuman, Monkey, Rat
IsotypeRabbit IgG
ImmunogenE.coli-derived human EYA4 recombinant protein (Position: R28-E205).
Antibody TypePrimary Antibody
ConcentrationAdding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
Form/AppearanceLyophilized
ConjugationUnconjugated
MW50 kDa
UniProt IDO95677
StorageMaintain 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 namesVascular endothelial growth factor B; VEGF-B; VEGF
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NoteFor research use only
Application notesWestern blot, 0.25-0.5μg/ml, Human, Monkey, Rat Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human ELISA, 0.1-0.5μg/ml, -. Add 0.2ml of distilled water will yield a concentration of 500ug/ml
Expiration Date12 months from date of receipt.
Anti-EYA4 Antibody

Flow Cytometry analysis of HELA cells using anti-EYA4 antibody. Overlay histogram showing HELA cells (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-EYA4 Antibody (1 µg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10 µg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 µg/1x10^6) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Anti-EYA4 Antibody

IHC analysis of EYA4 using anti-EYA4 antibody. EYA4 was detected in paraffin-embedded section of human mammary cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml rabbit anti-EYA4 Antibody overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

Anti-EYA4 Antibody

IHC analysis of EYA4 using anti-EYA4 antibody. EYA4 was detected in paraffin-embedded section of human rectal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml rabbit anti-EYA4 Antibody overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

Anti-EYA4 Antibody

Western blot analysis of EYA4 using anti-EYA4 antibody. Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50 ug of sample under reducing conditions. Lane 1: monkey skeletal muscle tissue lysates, Lane 2: human A431 whole cell lysates, Lane 3: human HT1080 whole cell lysates, Lane 4: rat skeletal muscle tissue lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-EYA4 antigen affinity purified polyclonal antibody at 0.5 µg/mL overnight at 4°C, then washed with TBS-0.1% Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for EYA4 at approximately 50 KD. The expected band size for EYA4 is at 50 KD.

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