FCM/FACS (Flow Cytometry)
(Figure 5. Flow Cytometry analysis of RT4 cells using anti-P3H2 antibody (AAA127575).Overlay histogram showing RT4 cells stained with AAA127575 (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-P3H2 Antibody (AAA127575, 1ug/1x106 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-rabbit IgG was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.)
Rabbit P3H2 Polyclonal Antibody | anti-P3H2 antibody
Anti-P3H2 Antibody Picoband
FCM/FACS (Flow Cytometry)
(Figure 5. Flow Cytometry analysis of RT4 cells using anti-P3H2 antibody (AAA127575).Overlay histogram showing RT4 cells stained with AAA127575 (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-P3H2 Antibody (AAA127575, 1ug/1x106 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-rabbit IgG was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.)
FCM/FACS (Flow Cytometry)
(Figure 5. Flow Cytometry analysis of RT4 cells using anti-P3H2 antibody (AAA127575).Overlay histogram showing RT4 cells stained with AAA127575 (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-P3H2 Antibody (AAA127575, 1ug/1x106 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-rabbit IgG was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.)
IF (Immunofluorescence)
(Figure 4. IF analysis of P3H2 using anti-P3H2 antibody (AAA127575) and anti-Beta Tubulin antibody (M01857-3).P3H2 was detected in immunocytochemical section of A549 cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5ug/mL rabbit anti-P3H2 Antibody (AAA127575) and mouse anti-Beta Tubulin antibody (M01857-3) overnight at 4 degree C. DyLight488 Conjugated Goat Anti-Rabbit IgG and DyLight594 Conjugated Goat Anti-Mouse IgG (BA1142) were used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37 degree C. Visualize using a fluorescence microscope and filter sets appropriate for the label used.)
IF (Immunofluorescence)
(Figure 4. IF analysis of P3H2 using anti-P3H2 antibody (AAA127575) and anti-Beta Tubulin antibody (M01857-3).P3H2 was detected in immunocytochemical section of A549 cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5ug/mL rabbit anti-P3H2 Antibody (AAA127575) and mouse anti-Beta Tubulin antibody (M01857-3) overnight at 4 degree C. DyLight488 Conjugated Goat Anti-Rabbit IgG and DyLight594 Conjugated Goat Anti-Mouse IgG (BA1142) were used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37 degree C. Visualize using a fluorescence microscope and filter sets appropriate for the label used.)
IHC (Immunohistochemisry)
(Figure 3. IHC analysis of P3H2 using anti-P3H2 antibody (AAA127575).P3H2 was detected in a paraffin-embedded section of rat kidney tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-P3H2 Antibody (AAA127575) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.)
IHC (Immunohistochemisry)
(Figure 3. IHC analysis of P3H2 using anti-P3H2 antibody (AAA127575).P3H2 was detected in a paraffin-embedded section of rat kidney tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-P3H2 Antibody (AAA127575) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.)
IHC (Immunohiostchemistry)
(Figure 2. IHC analysis of P3H2 using anti-P3H2 antibody (AAA127575).P3H2 was detected in a paraffin-embedded section of mouse kidney tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-P3H2 Antibody (AAA127575) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.)
IHC (Immunohiostchemistry)
(Figure 2. IHC analysis of P3H2 using anti-P3H2 antibody (AAA127575).P3H2 was detected in a paraffin-embedded section of mouse kidney tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-P3H2 Antibody (AAA127575) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.)
WB (Western Blot)
(Figure 1. Western blot analysis of P3H2 using anti-P3H2 antibody (AAA127575).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 Hela whole cell lysates,Lane 2: human RT4 whole cell lysates,Lane 3: human SIHA whole cell lysates,Lane 4: human A431 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-P3H2 antigen affinity purified polyclonal antibody (#AAA127575) at 0.5ug/mL overnight at 4 degree 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 P3H2 at approximately 81 kDa. The expected band size for P3H2 is at 81,60 kDa.)
WB (Western Blot)
(Figure 1. Western blot analysis of P3H2 using anti-P3H2 antibody (AAA127575).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 Hela whole cell lysates,Lane 2: human RT4 whole cell lysates,Lane 3: human SIHA whole cell lysates,Lane 4: human A431 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-P3H2 antigen affinity purified polyclonal antibody (#AAA127575) at 0.5ug/mL overnight at 4 degree 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 P3H2 at approximately 81 kDa. The expected band size for P3H2 is at 81,60 kDa.)
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Product Notes
The P3H2 leprel1 (Catalog #AAA127575) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-P3H2 Antibody Picoband reacts with Human, Mouse, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's P3H2 can be used in a range of immunoassay formats including, but not limited to, ELISA, FCM/FACS (Flow Cytometry), IF (Immunofluorescence), ICC (Immunocytochemistry), IHC (Immunohistochemistry), WB (Western Blot). Researchers should empirically determine the suitability of the P3H2 leprel1 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "P3H2, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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