CD2AP, Monoclonal Antibody

CD2-associated protein; Adapter protein CMS; Cas ligand with multiple SH3 domains; CD2AP; CD2 associated protein

For Research Use Only. Not for use in diagnostic procedures.

Monoclonal

IgG1

5F8

Mouse

No cross reactivity with other proteins.

Immunogen Affinity Purified

Lyophilized

Store at -20 degree C for one year. After reconstitution, at 4 degree C for one month. It can also be aliquotted and stored frozen at -20 degree C for a longer time.
Avoid repeated freezing and thawing.

Manufactured in an ISO 13485:2003 and EN ISO 13485:2012 Certified Laboratory.

Small volumes of anti-CD2AP antibody vial(s) may occasionally become entrapped in the seal of the product vial during shipment and storage. If necessary, briefly centrifuge the vial on a tabletop centrifuge to dislodge any liquid in the container`s cap. Certain products may require to ship with dry ice and additional dry ice fee may apply.

Description: Mouse IgG monoclonal antibody for CD2AP detection. Tested with WB, IHC-P, FCM in Human; Mouse; Rat.
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 s, SH3 domains, and a proline-rich region containing s 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.

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.

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.

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.

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.

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.

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.

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.

71,451 Da

CD2 associated protein

CD2-associated protein

CD2-associated protein

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 s, SH3 domains, and a proline-rich region containing s 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]

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).

1. Cochran, J. N., Rush, T., Buckingham, S. C., Roberson, E. D. The Alzheimer's disease risk factor CD2AP maintains blood-brain barrier integrity. Hum. Molec. Genet. 24: 6667-6674, 2015. 2. Lowik, M. M., Groenen, P. J. T. A., Pronk, I., Lilien, M. R., Goldschmeding, R., Dijkman, H. B., Levtchenko, E. N., Monnens, L. A., van den Heuvel, L. P. Focal segmental glomerulosclerosis in a patient homozygous for a CD2AP mutation. Kidney Int. 72: 1198-1203, 2007.

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