PHPT1 Rabbit Polyclonal Antibody

SKU: orb1972599

Description

Anti-PHPT1 Antibody. Tested in WB, IHC, Flow Cytometry, ELISA applications. This antibody reacts with Human, Mouse, Rat.

Images & Validation

• Tested Applications: ELISA, FC, IHC, WB
• Dilution Range: Western blot, 0.25-0.5 μg/ml, Human, Mouse, Rat Immunohistochemistry (Paraffin-embedded Section), 2-5 μg/ml, Human Flow Cytometry (Fixed), 1-3 μg/1x10^6 cells, Human ELISA, 0.1-0.5 μg/ml, -
• Reactivity: Human, Mouse, Rat

Key Properties

• Antibody Type: Primary Antibody
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: IgG
• Immunogen: E.coli-derived human PHPT1 recombinant protein (Position: M1-Y125). Human PHPT1 shares 84.4% amino acid (aa) sequence identity with mouse PHPT1.
• Target: 14 kDa phosphohistidine phosphatase
• Molecular Weight: 14 kDa
• Purification: Immunogen affinity purified.
• Conjugation: Unconjugated

Storage & Handling

• 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.
• Form/Appearance: Lyophilized
• Buffer/Preservatives: Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Expiration Date: 12 months from date of receipt.
• Disclaimer: For research use only

Alternative Names

PHPT1; PHP14; CGI-202; HSPC141; 14 kDa phosphohistidine phosphatase; Phosphohistidine phosphatase 1; PHPT1; Protein histidine phosphatase; PHP; Protein janus-A homolog

Flow Cytometry analysis of THP-1 cells using anti-PHPT1 antibody. Overlay histogram showing THP-1 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-PHPT1 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 (Red line) was also used as a control.

IHC analysis of PHPT1 using anti-PHPT1 antibody. PHPT1 was detected in a paraffin-embedded section of human skeletal muscle 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 2 µg/ml rabbit anti-PHPT1 Antibody overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.

IHC analysis of PHPT1 using anti-PHPT1 antibody. PHPT1 was detected in a paraffin-embedded section of human skeletal muscle 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 2 µg/ml rabbit anti-PHPT1 Antibody overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.

Western blot analysis of PHPT1 using anti-PHPT1 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 30 ug of sample under reducing conditions. Lane 1: human MCF-7 whole cell lysates, Lane 2: human A549 whole cell lysates, Lane 3: human THP-1 whole cell lysates, Lane 4: human 293T whole cell lysates, Lane 5: rat kidney tissue lysates, Lane 6: rat heart tissue lysates, Lane 7: mouse kidney tissue lysates, Lane 8: mouse heart 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-PHPT1 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 PHPT1 at approximately 14 kDa. The expected band size for PHPT1 is at 14 kDa.

Quick Database Links

Gene Symbol

14 kDa phosphohistidine phosphatase

UniProt

Q9NRX4