FCM/FACS (Flow Cytometry)
(Figure 3. Flow Cytometry analysis of HepG2 cells using anti-SPRED2 antibody (AAA127363).Overlay histogram showing HepG2 cells stained with AAA127363 (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-SPRED2 Antibody (AAA127363, 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 SPRED2 Polyclonal Antibody | anti-SPRED2 antibody
Anti-SPRED2 Antibody Picoband
FCM/FACS (Flow Cytometry)
(Figure 3. Flow Cytometry analysis of HepG2 cells using anti-SPRED2 antibody (AAA127363).Overlay histogram showing HepG2 cells stained with AAA127363 (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-SPRED2 Antibody (AAA127363, 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 3. Flow Cytometry analysis of HepG2 cells using anti-SPRED2 antibody (AAA127363).Overlay histogram showing HepG2 cells stained with AAA127363 (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-SPRED2 Antibody (AAA127363, 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 2. IF analysis of SPRED2 using anti-SPRED2 antibody (AAA127363) and anti-Tubulin Alpha antibody (M03989-3).SPRED2 was detected in immunocytochemical section of U20S 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-SPRED2 Antibody (AAA127363) and mouse anti-Tubulin Alpha antibody (M03989-3) overnight at 4 degree C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) and DyLight488 Conjugated Goat Anti-Mouse IgG (BA1126) 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 2. IF analysis of SPRED2 using anti-SPRED2 antibody (AAA127363) and anti-Tubulin Alpha antibody (M03989-3).SPRED2 was detected in immunocytochemical section of U20S 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-SPRED2 Antibody (AAA127363) and mouse anti-Tubulin Alpha antibody (M03989-3) overnight at 4 degree C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) and DyLight488 Conjugated Goat Anti-Mouse IgG (BA1126) 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.)
WB (Western Blot)
(Figure 1. Western blot analysis of SPRED2 using anti-SPRED2 antibody (AAA127363).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: rat brain tissue lysates,Lane 2: mouse brain 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-SPRED2 antigen affinity purified polyclonal antibody (#AAA127363) 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 SPRED2 at approximately 53 kDa. The expected band size for SPRED2 is at 48 kDa.)
WB (Western Blot)
(Figure 1. Western blot analysis of SPRED2 using anti-SPRED2 antibody (AAA127363).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: rat brain tissue lysates,Lane 2: mouse brain 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-SPRED2 antigen affinity purified polyclonal antibody (#AAA127363) 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 SPRED2 at approximately 53 kDa. The expected band size for SPRED2 is at 48 kDa.)
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Product Notes
The SPRED2 spred2 (Catalog #AAA127363) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-SPRED2 Antibody Picoband reacts with Human, Mouse, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's SPRED2 can be used in a range of immunoassay formats including, but not limited to, ELISA, FCM/FACS (Flow Cytometry), IF (Immunofluorescence), ICC (Immunocytochemistry), WB (Western Blot). Researchers should empirically determine the suitability of the SPRED2 spred2 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 "SPRED2, 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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