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Catalog Number | orb654428 |
---|---|
Category | Antibodies |
Description | Glypican 5/GPC5 Antibody |
Species/Host | Rabbit |
Clonality | Polyclonal |
Tested applications | ELISA, FC, ICC, IF, WB |
Reactivity | Human |
Isotype | Rabbit IgG |
Immunogen | E.coli-derived human Glypican 5/GPC5 recombinant protein (Position: Q28-D538). |
Concentration | Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml. |
Form/Appearance | Lyophilized |
Conjugation | Unconjugated |
MW | 64 kDa |
UniProt ID | P78333 |
Storage | Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles. |
Alternative names | Formin-1;Limb deformity protein homolog;FMN1;FMN, Read more... |
Note | For research use only |
Application notes | Tested Species: In-house tested species with positive results. Other applications have not been tested. Optimal dilutions should be determined by end users. Add 0.2ml of distilled water will yield a concentration of 500ug/ml. |
Expiration Date | 12 months from date of receipt. |
Western blot analysis of Glypican 5/GPC5 using anti-Glypican 5/GPC5 antibody (orb654428). 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 50ug of sample under reducing conditions. Lane 1: human HeLa whole cell lysates, Lane 2: human U87 whole cell lysates, Lane 3: human U20S whole cell lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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-Glypican 5/GPC5 antigen affinity purified polyclonal antibody (Catalog # orb654428) 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 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # orb90503) with Tanon 5200 system. A specific band was detected for Glypican 5/GPC5 at approximately 64KD. The expected band size for Glypican 5/GPC5 is at 64KD.
Flow Cytometry analysis of U20S cells using anti-Glypican 5/GPC5 antibody (orb654428). Overlay histogram showing U20S cells stained with orb654428 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Glypican 5/GPC5 Antibody (orb654428, 1μg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
Flow Cytometry analysis of U87 cells using anti-Glypican 5/GPC5 antibody (orb654428). Overlay histogram showing U87 cells stained with orb654428 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Glypican 5/GPC5 Antibody (orb654428, 1μg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
IF analysis of Glypican 5/GPC5 using anti-Glypican 5/GPC5 antibody (orb654428). Glypican 5/GPC5 was detected in immunocytochemical section of A431 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (orb90553) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 4μg/mL rabbit anti-Glypican 5/GPC5 Antibody (orb654428) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
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