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Catalog Number | orb570361 |
---|---|
Category | Antibodies |
Description | Anti-TRPV5 Antibody. Tested in ELISA, Flow Cytometry, IHC, IHC-F, ICC, WB applications. This antibody reacts with Human, Mouse, Rat. |
Species/Host | Rabbit |
Clonality | Polyclonal |
Tested applications | ELISA, FC, ICC, IHC, IHC-Fr, WB |
Reactivity | Human, Mouse, Rat |
Isotype | Rabbit IgG |
Immunogen | E.coli-derived human TRPV5 recombinant protein (Position: Q14-Q74+H636-F729). |
Concentration | Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml. |
Form/Appearance | Lyophilized |
Conjugation | Unconjugated |
MW | 83 kDa |
UniProt ID | Q9NQA5 |
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 | Transient receptor potential cation channel subfam Read more... |
Note | For research use only |
Application notes | Western blot, 0.25-0.5μg/ml Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml Immunohistochemistry (Frozen Section), 0.5-1μg/ml Immunocytochemistry, 0.5-1μg/ml Flow Cytometry (Fixed), 1-3μg/1x106 cells ELISA, 0.1-0.5μg/ml. Add 0.2ml of distilled water will yield a concentration of 500ug/ml |
Expiration Date | 12 months from date of receipt. |
Flow Cytometry analysis of HepG2 cells using anti-TRPV5 antibody. Overlay histogram showing HepG2 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-TRPV5 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.
Flow Cytometry analysis of U87 cells using anti-TRPV5 antibody. Overlay histogram showing U87 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-TRPV5 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.
IHC analysis of TRPV5 using anti-TRPV5 antibody. TRPV5 was detected in paraffin-embedded section of human pancreatic cancer tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml rabbit anti-TRPV5 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.
IHC analysis of TRPV5 using anti-TRPV5 antibody. TRPV5 was detected in paraffin-embedded section of mouse kidney tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml rabbit anti-TRPV5 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.
IHC analysis of TRPV5 using anti-TRPV5 antibody. TRPV5 was detected in paraffin-embedded section of rat kidney tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml rabbit anti-TRPV5 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.
Western blot analysis of TRPV5 using anti-TRPV5 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 COS-7 whole cell lysates, Lane 2: human U-87MG whole cell lysates, Lane 3: human Hela whole cell lysates, Lane 4: human HepG2 whole cell 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-TRPV5 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 1:10000 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 TRPV5 at approximately 83 KD. The expected band size for TRPV5 is at 83 KD.
ELISA, ICC, IF, WB | |
Bovine, Canine, Human, Mouse, Porcine, Rat | |
Goat | |
Polyclonal | |
Unconjugated |
ELISA, ICC, IF, WB | |
Bovine, Canine, Human, Mouse, Porcine, Rat | |
Goat | |
Polyclonal | |
Unconjugated |
IF, WB | |
Human, Mouse, Rat | |
Rabbit | |
Polyclonal | |
Unconjugated |