Western Blot (WB)
(Figure 1. Western blot analysis of ADA using anti-ADA antibody (MBS1752027). 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 Hela whole cell lysates, Lane 2: human placenta tissue lysates, Lane 3: human A549 whole cell lysates, Lane 4: human MCF-7 whole cell lysates, Lane 5: human U-937 whole cell lysates, Lane 6: human U20S whole cell lysates, Lane 7: human Caco-2 whole cell lysates. 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-ADA 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 anti-Human ADA Monoclonal Antibody | anti-ADA antibody
Anti-ADA Antibody (monoclonal, 6D4)
IHC-P:0.5-1ug/ml; Antigen Retrieval: By heat
(FC): 1-3ug/1x106 cells.
Tested Species: In-house tested species with positice results.
By heat: Boiling the paraffin section 10mM citrate buffer, pH6.0, for 20mins is required for staining of formalin.paraffin sections.
Other applications have not been tested.
Optimal dilutions should be determined by end users.
Enhanced Chemiluminescent Kit with anti-Mouse IgG (MBS176445) for Western blot, and HRP Conjugated anti-Mouse IgG Super Vision Assay Kit (MBS176472) for IHC(P).
Avoid repeated freezing and thawing.
Western Blot (WB)
(Figure 1. Western blot analysis of ADA using anti-ADA antibody (MBS1752027). 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 Hela whole cell lysates, Lane 2: human placenta tissue lysates, Lane 3: human A549 whole cell lysates, Lane 4: human MCF-7 whole cell lysates, Lane 5: human U-937 whole cell lysates, Lane 6: human U20S whole cell lysates, Lane 7: human Caco-2 whole cell lysates. 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-ADA 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 ADA using anti-ADA antibody (MBS1752027). 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 Hela whole cell lysates, Lane 2: human placenta tissue lysates, Lane 3: human A549 whole cell lysates, Lane 4: human MCF-7 whole cell lysates, Lane 5: human U-937 whole cell lysates, Lane 6: human U20S whole cell lysates, Lane 7: human Caco-2 whole cell lysates. 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-ADA 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 ADA using anti-ADA antibody (MBS1752027). ADA was detected in paraffin-embedded section of human colon cancer 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-ADA Antibody (MBS1752027) 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 ADA using anti-ADA antibody (MBS1752027). ADA was detected in paraffin-embedded section of human colon cancer 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-ADA Antibody (MBS1752027) 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 ADA using anti-ADA antibody (MBS1752027). ADA was detected in paraffin-embedded section of human colon cancer 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-ADA Antibody (MBS1752027) 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 ADA using anti-ADA antibody (MBS1752027). ADA was detected in paraffin-embedded section of human colon cancer 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-ADA Antibody (MBS1752027) 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 ADA using anti-ADA antibody (MBS1752027). ADA was detected in paraffin-embedded section of human colon cancer 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-ADA Antibody (MBS1752027) 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 ADA using anti-ADA antibody (MBS1752027). ADA was detected in paraffin-embedded section of human colon cancer 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-ADA Antibody (MBS1752027) 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 ADA using anti-ADA antibody (MBS1752027). ADA was detected in paraffin-embedded section of human rectal cancer 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-ADA Antibody (MBS1752027) 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 ADA using anti-ADA antibody (MBS1752027). ADA was detected in paraffin-embedded section of human rectal cancer 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-ADA Antibody (MBS1752027) 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 6. IHC analysis of ADA using anti-ADA antibody (MBS1752027). ADA was detected in paraffin-embedded section of human tonsil 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-ADA Antibody (MBS1752027) 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 6. IHC analysis of ADA using anti-ADA antibody (MBS1752027). ADA was detected in paraffin-embedded section of human tonsil 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-ADA Antibody (MBS1752027) 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 7. Flow Cytometry analysis of U20S cells using anti- ADA antibody (MBS1752027). Overlay histogram showing U20S cells stained with MBS1752027 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with mouse anti- ADA Antibody (MBS1752027, 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 7. Flow Cytometry analysis of U20S cells using anti- ADA antibody (MBS1752027). Overlay histogram showing U20S cells stained with MBS1752027 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with mouse anti- ADA Antibody (MBS1752027, 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: Adenosine Deaminase (also known as adenosine aminohydrolase, or ADA) is an enzyme involved in purine metabolism. Primarily, ADA in humans is involved in the development and maintenance of the immune system. However, ADA association has also been observed with epithelial cell differentiation, neurotransmission, and gestation maintenance. It has also been proposed that ADA, in addition to adenosine breakdown, stimulates release of excitatory amino acids and is necessary to the coupling of A1 adenosine receptors and heterotrimeric G proteins. Adenosine deaminase deficiency leads to pulmonary fibrosis, suggesting that chronic exposure to high levels of adenosine can exacerbate inflammation responses rather than suppressing them. It has also been recognized that adenosine deaminase protein and activity is upregulated in mouse hearts that overexpress HIF-1 alpha, which in part explains the attenuated levels of adenosine in HIF-1 alpha expressing hearts during ischemic stress.
NCBI and Uniprot Product Information
NCBI Description
This gene encodes an enzyme that catalyzes the hydrolysis of adenosine to inosine. Various mutations have been described for this gene and have been linked to human diseases. Deficiency in this enzyme causes a form of severe combined immunodeficiency disease (SCID), in which there is dysfunction of both B and T lymphocytes with impaired cellular immunity and decreased production of immunoglobulins, whereas elevated levels of this enzyme have been associated with congenital hemolytic anemia. [provided by RefSeq, Jul 2008]
Uniprot Description
Catalyzes the hydrolytic deamination of adenosine and 2-deoxyadenosine (PubMed:8452534, PubMed:16670267). Plays an important role in purine metabolism and in adenosine homeostasis. Modulates signaling by extracellular adenosine, and so contributes indirectly to cellular signaling events. Acts as a positive regulator of T-cell coactivation, by binding DPP4 (PubMed:20959412). Its interaction with DPP4 regulates lymphocyte-epithelial cell adhesion (PubMed:11772392). Enhances dendritic cell immunogenicity by affecting dendritic cell costimulatory molecule expression and cytokines and chemokines secretion (). Enhances CD4+ T-cell differentiation and proliferation (PubMed:20959412). Acts as a positive modulator of adenosine receptors ADORA1 and ADORA2A, by enhancing their ligand affinity via conformational change (PubMed:23193172). Stimulates plasminogen activation (PubMed:15016824). Plays a role in male fertility (PubMed:21919946, PubMed:26166670). Plays a protective role in early postimplantation embryonic development ().
Research Articles on ADA
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Product Notes
The ADA ada (Catalog #AAA1752027) is an Antibody produced from Mouse and is intended for research purposes only. The product is available for immediate purchase. The Anti-ADA Antibody (monoclonal, 6D4) reacts with Human and may cross-react with other species as described in the data sheet. AAA Biotech's ADA 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: 0.1-0.5ug/ml IHC-P:0.5-1ug/ml; Antigen Retrieval: By heat (FC): 1-3ug/1x106 cells. Tested Species: In-house tested species with positice results. By heat: Boiling the paraffin section 10mM citrate buffer, pH6.0, for 20mins is required for staining of formalin.paraffin sections. Other applications have not been tested. Optimal dilutions should be determined by end users. Enhanced Chemiluminescent Kit with anti-Mouse IgG (MBS176445) for Western blot, and HRP Conjugated anti-Mouse IgG Super Vision Assay Kit (MBS176472) for IHC(P). Researchers should empirically determine the suitability of the ADA ada 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 "ADA, 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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