Western Blot (WB)
(Figure 1. Western blot analysis of D2AP using anti-D2AP antibody (M01756). 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 K562 whole cell lysate, Lane 2: human A431 whole cell lysate, Lane 3: human 293T whole cell lysate, Lane 4: human U20S whole cell lysate, Lane 5: human HL-60 whole cell lysate, Lane 6: human MCF-7 whole cell lysate, Lane 7: human Hela whole cell lysate, Lane 8: human PANC-1 whole cell lysate. 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 mouse anti-D2AP antigen affinity purified monoclonal antibody at 0.5 ug/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-mouse 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.)
Mouse CD2AP Monoclonal Antibody | anti-CD2AP antibody
Anti-CD2AP Antibody (monoclonal, 5F8)
IHC-P: 0.5-1 mug/ml
FC/FACS: 1-3ug/1x106 cells
Tested Species: In-house tested species with positive results. By Heat: Boiling the paraffin sections in 10mM citrate buffer, pH6.0, for 20mins is required for the staining of formalin/paraffin sections.
Avoid repeated freezing and thawing.
Western Blot (WB)
(Figure 1. Western blot analysis of D2AP using anti-D2AP antibody (M01756). 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 K562 whole cell lysate, Lane 2: human A431 whole cell lysate, Lane 3: human 293T whole cell lysate, Lane 4: human U20S whole cell lysate, Lane 5: human HL-60 whole cell lysate, Lane 6: human MCF-7 whole cell lysate, Lane 7: human Hela whole cell lysate, Lane 8: human PANC-1 whole cell lysate. 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 mouse anti-D2AP antigen affinity purified monoclonal antibody at 0.5 ug/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-mouse 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.)
Western Blot (WB)
(Figure 1. Western blot analysis of D2AP using anti-D2AP antibody (M01756). 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 K562 whole cell lysate, Lane 2: human A431 whole cell lysate, Lane 3: human 293T whole cell lysate, Lane 4: human U20S whole cell lysate, Lane 5: human HL-60 whole cell lysate, Lane 6: human MCF-7 whole cell lysate, Lane 7: human Hela whole cell lysate, Lane 8: human PANC-1 whole cell lysate. 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 mouse anti-D2AP antigen affinity purified monoclonal antibody at 0.5 ug/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-mouse 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.)
Immunohistochemistry (IHC)
(Figure 3. IHC analysis of CD2AP using anti-CD2AP antibody (M01756). CD2AP was detected in paraffin-embedded section of human colon cancer. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epi1ope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml mouse anti-CD2AP Antibody (M01756) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 3. IHC analysis of CD2AP using anti-CD2AP antibody (M01756). CD2AP was detected in paraffin-embedded section of human colon cancer. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epi1ope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml mouse anti-CD2AP Antibody (M01756) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 3. IHC analysis of CD2AP using anti-CD2AP antibody (M01756). CD2AP was detected in paraffin-embedded section of human colon cancer. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epi1ope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml mouse anti-CD2AP Antibody (M01756) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 3. IHC analysis of CD2AP using anti-CD2AP antibody (M01756). CD2AP was detected in paraffin-embedded section of human colon cancer. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epi1ope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml mouse anti-CD2AP Antibody (M01756) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 4. IHC analysis of CD2AP using anti-CD2AP antibody (M01756). CD2AP was detected in paraffin-embedded section of human placenta tissue. 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 1ug/ml mouse anti-CD2AP Antibody (M01756) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 4. IHC analysis of CD2AP using anti-CD2AP antibody (M01756). CD2AP was detected in paraffin-embedded section of human placenta tissue. 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 1ug/ml mouse anti-CD2AP Antibody (M01756) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 5. IHC analysis of CD2AP using anti-CD2AP antibody (M01756). CD2AP was detected in paraffin-embedded section of human mammary cancer. 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 1ug/ml mouse anti-CD2AP Antibody (M01756) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 5. IHC analysis of CD2AP using anti-CD2AP antibody (M01756). CD2AP was detected in paraffin-embedded section of human mammary cancer. 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 1ug/ml mouse anti-CD2AP Antibody (M01756) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen. )
Flow Cytometry (FC/FACS)
(Figure 6. Flow Cytometry analysis of K562 cells using anti-D2AP antibody (M01756). Overlay histogram showing K562 cells stained with M01756 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-D2AP Antibody (M01756, 1ug/1x106 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-mouse IgG (BA1126, 5-10ug/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was mouse IgG (1ug/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)
Flow Cytometry (FC/FACS)
(Figure 6. Flow Cytometry analysis of K562 cells using anti-D2AP antibody (M01756). Overlay histogram showing K562 cells stained with M01756 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-D2AP Antibody (M01756, 1ug/1x106 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-mouse IgG (BA1126, 5-10ug/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was mouse IgG (1ug/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)
Background: CD2-associated protein is a protein that in humans is encoded by the CD2AP gene. This gene encodes a scaffolding molecule that regulates the actin cytoskeleton. The protein directly interacts with filamentous actin and a variety of cell membrane proteins through multiple actin binding sites, SH3 domains, and a proline-rich region containing binding sites for SH3 domains. The cytoplasmic protein localizes to membrane ruffles, lipid rafts, and the leading edges of cells. It is implicated in dynamic actin remodeling and membrane trafficking that occurs during receptor endocytosis and cytokinesis. Haploinsufficiency of this gene is implicated in susceptibility to glomerular disease.
NCBI and Uniprot Product Information
NCBI Description
This gene encodes a scaffolding molecule that regulates the actin cytoskeleton. The protein directly interacts with filamentous actin and a variety of cell membrane proteins through multiple actin binding sites, SH3 domains, and a proline-rich region containing binding sites for SH3 domains. The cytoplasmic protein localizes to membrane ruffles, lipid rafts, and the leading edges of cells. It is implicated in dynamic actin remodeling and membrane trafficking that occurs during receptor endocytosis and cytokinesis. Haploinsufficiency of this gene is implicated in susceptibility to glomerular disease. [provided by RefSeq, Jul 2008]
Uniprot Description
Seems to act as an adapter protein between membrane proteins and the actin cytoskeleton (PubMed:10339567). In collaboration with CBLC, modulates the rate of RET turnover and may act as regulatory checkpoint that limits the potency of GDNF on neuronal survival. Controls CBLC function, converting it from an inhibitor to a promoter of RET degradation (). May play a role in receptor clustering and cytoskeletal polarity in the junction between T-cell and antigen-presenting cell (). May anchor the podocyte slit diaphragm to the actin cytoskeleton in renal glomerolus. Also required for cytokinesis (PubMed:15800069). Plays a role in epithelial cell junctions formation (PubMed:22891260).
Research Articles on CD2AP
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Product Notes
The CD2AP cd2ap (Catalog #AAA1752130) is an Antibody produced from Mouse and is intended for research purposes only. The product is available for immediate purchase. The Anti-CD2AP Antibody (monoclonal, 5F8) reacts with Human, Mouse, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's CD2AP can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Immunohistochemistry (IHC) Paraffin, Flow Cytometry (FC/FACS). WB: 0.1-0.5 mug/ml IHC-P: 0.5-1 mug/ml FC/FACS: 1-3ug/1x106 cells Tested Species: In-house tested species with positive results. By Heat: Boiling the paraffin sections in 10mM citrate buffer, pH6.0, for 20mins is required for the staining of formalin/paraffin sections. Researchers should empirically determine the suitability of the CD2AP cd2ap 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 "CD2AP, Monoclonal 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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