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Product Info Summary
| SKU: | M01756 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Mouse |
| Application: | Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-CD2AP Antibody Picoband® (monoclonal, 5F8)
SKU/Catalog Number
M01756
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-CD2AP Antibody Picoband® (monoclonal, 5F8) catalog # M01756. Tested in Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse, Rat. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
Storage & Handling
Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.
Cite This Product
Anti-CD2AP Antibody Picoband® (monoclonal, 5F8) (Boster Biological Technology, Pleasanton CA, USA, Catalog # M01756)
Host
Mouse
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clonality
Monoclonal
Clone Number
5F8
Isotype
Mouse IgG1
Immunogen
E. coli-derived human CD2AP recombinant protein (Position: K253-K337).
*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
M01756 is reactive to CD2AP in Human, Mouse, Rat
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500μg/ml.
Observed Molecular Weight
80 kDa
Calculated molecular weight
70.45kDa
Background of Cd2ap
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.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Application & Images
Applications
M01756 is guaranteed for Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.1-0.5μg/ml
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml
Immunocytochemistry/Immunofluorescence, 5 μg/ml
Flow Cytometry (Fixed), 1-3μg/1x106 cells
Positive Control
WB: human K562 whole cell, human A431 whole cell, human HEK293 whole cell, human U20S whole cell, human HL-60 whole cell, human MCF-7 whole cell, human Hela whole cell, human PANC-1 whole cell,, rat liver tissue, rat testicular tissue, mouse pancreas tissue, mouse testicular tissue, mouse RAW2467 whole cell
IHC: human mammary cancer, human mammary cancer, human colon cancer, human placenta tissue, mouse kidney tissue, rat brain tissue
ICC: A431 cell
FCM: K562 cell, U20S cell
Validation Images & Assay Conditions
Click image to see more details
Figure 1. Western blot analysis of CD2AP using anti-CD2AP 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 HEK293 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-CD2AP antigen affinity purified monoclonal antibody (Catalog # M01756) 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-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 (Catalog # EK1001) with Tanon 5200 system. A specific band was detected for CD2AP at approximately 80KD. The expected band size for CD2AP is at 71KD.
Click image to see more details
Figure 2. 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, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-CD2AP Antibody (M01756) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1021) with DAB as the chromogen.
Click image to see more details
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 1μg/ml mouse anti-CD2AP Antibody (M01756) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1021) with DAB as the chromogen.
Click image to see more details
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 1μg/ml mouse anti-CD2AP Antibody (M01756) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1021) with DAB as the chromogen.
Click image to see more details
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 1μg/ml mouse anti-CD2AP Antibody (M01756) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1021) with DAB as the chromogen.
Click image to see more details
Figure 6. Flow Cytometry analysis of K562 cells using anti-D2AP antibody (M01756).
Overlay histogram showing K562 cells stained with M01756 (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 mouse anti-D2AP Antibody (M01756,1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
Click image to see more details
Figure 7. 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 1μg/ml mouse anti-CD2AP Antibody (M01756) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1021) with DAB as the chromogen.
Click image to see more details
Figure 8. 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, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-CD2AP Antibody (M01756) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1021) with DAB as the chromogen.
Click image to see more details
Figure 9. 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, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-CD2AP Antibody (M01756) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1021) with DAB as the chromogen.
Click image to see more details
Figure 10. 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 1μg/ml mouse anti-CD2AP Antibody (M01756) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1021) with DAB as the chromogen.
Click image to see more details
Figure 11. IHC analysis of CD2AP using anti-CD2AP antibody (M01756).
CD2AP was detected in paraffin-embedded section of mouse kidney 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 1μg/ml mouse anti-CD2AP Antibody (M01756) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1021) with DAB as the chromogen.
Click image to see more details
Figure 12. IHC analysis of CD2AP using anti-CD2AP antibody (M01756).
CD2AP was detected in paraffin-embedded section of rat brain 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 1μg/ml mouse anti-CD2AP Antibody (M01756) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1021) with DAB as the chromogen.
Click image to see more details
Figure 13. IHC analysis of CD2AP using anti-CD2AP antibody (M01756).
CD2AP was detected in paraffin-embedded section of rat kidney 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 1μg/ml mouse anti-CD2AP Antibody (M01756) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1021) with DAB as the chromogen.
Click image to see more details
Figure 14. ICC analysis of CD2AP using anti-CD2AP antibody (M01756).
CD2AP was detected in immunocytochemical section of A431 cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 1μg/ml mouse anti-CD2AP Antibody (M01756) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1021) with DAB as the chromogen.
Click image to see more details
Figure 15. Flow Cytometry analysis of U20S cells using anti-CD2AP antibody (M01756).
Overlay histogram showing U20S cells stained with M01756 (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 mouse anti-CD2AP Antibody (M01756,1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
Click image to see more details
Figure 16. Western blot analysis of CD2AP using anti-CD2AP 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: rat liver tissue lysate,
Lane 2: rat testicular tissue lysate,
Lane 3: mouse pancreas tissue lysate,
Lane 4: mouse testicular tissue lysate,
Lane 5: mouse RAW246.7 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-CD2AP antigen affinity purified monoclonal antibody (Catalog # M01756) 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-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 (Catalog # EK1001) with Tanon 5200 system. A specific band was detected for CD2AP at approximately 80KD. The expected band size for CD2AP is at 71KD.
Protein Target Info & Infographic
Gene/Protein Information For CD2AP (Source: Uniprot.org, NCBI)
Gene Name
CD2AP
Full Name
CD2-associated protein
Weight
70.45kDa
Alternative Names
CD2-associated protein; Adapter protein CMS; Cas ligand with multiple SH3 domains; CD2AP Cd2ap|AL024079, C78928, METS-, METS-1, Mets, Mets1|CD2-associated protein|CD2-associated protein|SH3 domain-containing adapter protein|SH3 domain-containing adaptor protein METS-1|mesenchyme to epithelium transition protein, SH3 domains|mesenchyme-to-epithelium transition protein with SH3 domains 1
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on CD2AP, check out the CD2AP Infographic
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for CD2AP: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].
Specific Publications For Anti-CD2AP Antibody Picoband® (monoclonal, 5F8) (M01756)
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1 Customer Q&As for Anti-CD2AP Antibody Picoband® (monoclonal, 5F8)
Question
We are currently using anti-CD2AP antibody (monoclonal, 5F8) M01756 for mouse tissue, and we are content with the Flow Cytometry results. The species of reactivity given in the datasheet says human, mouse, rat. Is it likely that the antibody can work on bovine tissues as well?
Verified Customer
Verified customer
Asked: 2019-07-25
Answer
The anti-CD2AP antibody (monoclonal, 5F8) (M01756) has not been tested for cross reactivity specifically with bovine tissues, though there is a good chance of cross reactivity. We have an innovator award program that if you test this antibody and show it works in bovine you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2019-07-25
