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Rabbit Monoclonal ADA antibody. Carrier free. Suitable for WB, ICC/IF, IHC-P, IP and reacts with Human samples.

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Images

Immunoprecipitation - Anti-ADA antibody [EPR25429-117] (BSA and Azide free) (AB300051), expandable thumbnail
  • Western blot - Anti-ADA antibody [EPR25429-117] (BSA and Azide free) (AB300051), expandable thumbnail
  • Western blot - Anti-ADA antibody [EPR25429-117] (BSA and Azide free) (AB300051), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ADA antibody [EPR25429-117] (BSA and Azide free) (AB300051), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ADA antibody [EPR25429-117] (BSA and Azide free) (AB300051), expandable thumbnail

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Constituents: 100% PBS

Form
Liquid
Clonality
Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
WBICC/IFIHC-PIPFlow Cyt (Intra)
Human
Tested
Tested
Tested
Tested
Not recommended
Mouse
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended
Rat
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended

Tested
Tested

Species
Human
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Mouse, Rat
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Mouse, Rat
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Mouse, Rat
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Mouse, Rat
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Mouse, Human, Rat
Dilution info
-
Notes

-

Target data

Function

Catalyzes the hydrolytic deamination of adenosine and 2-deoxyadenosine (PubMed:16670267, PubMed:23193172, PubMed:26166670, PubMed:8452534, PubMed:9361033). 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 (By similarity). 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 (By similarity). Also responsible for the deamination of cordycepin (3'-deoxyadenosine), a fungal natural product that shows antitumor, antibacterial, antifungal, antivirus, and immune regulation properties (PubMed:26038697).

Alternative names

ADA1, ADA, Adenosine deaminase, Adenosine aminohydrolase

Recommended products

Rabbit Monoclonal ADA antibody. Carrier free. Suitable for WB, ICC/IF, IHC-P, IP and reacts with Human samples.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Carrier free
Yes
Clone number
EPR25429-117
Purification technique
Affinity purification Protein A
Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Aliquoting information
Upon delivery aliquot

Notes

ab300051 is a carrier free version of Anti-ADA antibody [EPR25429-117] ab300050.

Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

Adenosine deaminase (ADA) sometimes known as deaminase is an enzyme that plays a critical mechanical role in cellular metabolism. This enzyme catalyzes the deamination of adenosine converting it into inosine. ADA is a 41 kDa protein primarily expressed in most tissues including the thymus and spleen. Various ADA assays and ADA protocols exist to study its functionality and quantification often using ADA ELISA kits to measure ADA activity in biological samples. In cell lines such as HeLa ADA expression is also present and significant for studying its regulation and function.

Biological function summary

Adenosine deaminase activity influences purine metabolism playing an important role in the breakdown of adenosine. It is not part of a complex but interacts closely with other components of the purine salvage pathway to maintain nucleotide balance. This enzyme is important for the proper function of immune cells as it prevents toxic accumulations of adenosine which can be detrimental to lymphocyte proliferation and function. ADA's activity ensures the proper management of nucleotides within cells sustaining cellular health and functionality.

Pathways

Adenosine deaminase plays significant roles in the immune and purine catabolism pathways. Within these pathways ADA is closely related to proteins such as purine nucleoside phosphorylase (PNP) which together orchestrate the steps necessary for adenine and guanine nucleotide turnover. These proteins facilitate the conversion processes needed for cellular nucleic acid synthesis and energy regulation. By managing adenosine levels ADA's function ensures the smooth operation of cellular signaling and metabolic balance.

Associated diseases and disorders

Adenosine deaminase deficiency is linked to severe combined immunodeficiency (SCID) and can also be associated with pulmonary disorders like asthma. A lack of ADA activity leads to the accumulation of toxic metabolites impairing immune function and contributing to the pathogenesis of SCID. In some patients with pulmonary disorders altered ADA activity impacts the inflammatory response. ADA's mechanistic interactions with other proteins such as PNP highlight its importance in maintaining immune competence and metabolic homeostasis emphasizing the enzyme's role in these clinical conditions.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

9 product images

  • Immunoprecipitation - Anti-ADA antibody [EPR25429-117] (BSA and Azide free) (ab300051), expandable thumbnail

    Immunoprecipitation - Anti-ADA antibody [EPR25429-117] (BSA and Azide free) (ab300051)

    This data was developed using Anti-ADA antibody [EPR25429-117] ab300050, the same antibody clone in a different buffer formulation.

    ADA was immunoprecipitated from 0.35 mg Jurkat (human T cell leukemia T lymphocyte) whole cell lysate 10 µg with Anti-ADA antibody [EPR25429-117] ab300050 at 1/30 dilution. Western blot was performed on the immunoprecipitate using Anti-ADA antibody [EPR25429-117] ab300050 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.

    Lane 1: Jurkat (human T cell leukemia T lymphocyte) whole cell lysate 10 µg (Input)

    Lane 2: Anti-ADA antibody [EPR25429-117] ab300050 IP in Jurkat whole cell lysate

    Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-ADA antibody [EPR25429-117] ab300050 in Jurkat whole cell lysate

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    All lanes: Immunoprecipitation - Anti-ADA antibody [EPR25429-117] (Anti-ADA antibody [EPR25429-117] ab300050) at 1/1000 dilution

    Lane 1: Jurkat (human T cell leukemia T lymphocyte) whole cell lysate (Input) at 10 µg

    Lane 2: Anti-ADA antibody [EPR25429-117] ab300050 IP in Jurkat whole cell lysate

    Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-ADA antibody [EPR25429-117] ab300050 in Jurkat whole cell lysate

    Secondary

    All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution

    Predicted band size: 40 kDa

    Exposure time: 10s

  • Western blot - Anti-ADA antibody [EPR25429-117] (BSA and Azide free) (ab300051), expandable thumbnail

    Western blot - Anti-ADA antibody [EPR25429-117] (BSA and Azide free) (ab300051)

    This data was developed using Anti-ADA antibody [EPR25429-117] ab300050, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time:

    Lane1-2: 1 second;

    Lane3-4: 3.25 seconds.

    Low expression: human liver(PMID: 2606352)

    All lanes: Western blot - Anti-ADA antibody [EPR25429-117] (Anti-ADA antibody [EPR25429-117] ab300050) at 1/1000 dilution

    Lane 1: Jurkat (human T cell leukemia T lymphocyte) whole cell lysate at 20 µg

    Lane 2: MOLT-4 (human lymphoblastic leukemia T lymphoblast) whole cell lysate at 20 µg

    Lane 3: Human stomach tissue lysate at 20 µg

    Lane 4: Human liver tissue lysate at 20 µg

    Secondary

    All lanes: Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

    Predicted band size: 40 kDa

    Observed band size: 41 kDa, 36 kDa

  • Western blot - Anti-ADA antibody [EPR25429-117] (BSA and Azide free) (ab300051), expandable thumbnail

    Western blot - Anti-ADA antibody [EPR25429-117] (BSA and Azide free) (ab300051)

    This data was developed using Anti-ADA antibody [EPR25429-117] ab300050, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time:

    Lane1: 3.25 second;

    Lane2: 37 seconds.

    All lanes: Western blot - Anti-ADA antibody [EPR25429-117] (Anti-ADA antibody [EPR25429-117] ab300050) at 1/1000 dilution

    Lane 1: Human thymus tissue lysate at 20 µg

    Lane 2: Human tonsil tissue lysate at 20 µg

    Secondary

    All lanes: Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

    Predicted band size: 40 kDa

    Observed band size: 41 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ADA antibody [EPR25429-117] (BSA and Azide free) (ab300051), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ADA antibody [EPR25429-117] (BSA and Azide free) (ab300051)

    This data was developed using Anti-ADA antibody [EPR25429-117] ab300050, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded human stomach tissue labeling ADA with Anti-ADA antibody [EPR25429-117] ab300050 at 1/2000 followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection) was used. Positive staining on human stomach. The section was incubated with Anti-ADA antibody [EPR25429-117] ab300050 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: PBS was used instead of primary antibody (Anti-ADA antibody [EPR25429-117] ab300050) followed by ready to use secondary antibody LeicaDS9800 (BOND™ Polymer Refine Detection).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ADA antibody [EPR25429-117] (BSA and Azide free) (ab300051), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ADA antibody [EPR25429-117] (BSA and Azide free) (ab300051)

    This data was developed using Anti-ADA antibody [EPR25429-117] ab300050, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded human thymus tissue labelling ADA with Anti-ADA antibody [EPR25429-117] ab300050 at 1/2000 followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Positive staining on human thymus (PMID: 2606352). The section was incubated with Anti-ADA antibody [EPR25429-117] ab300050 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: PBS was used instead of primary antibody (Anti-ADA antibody [EPR25429-117] ab300050) followed by ready to use secondary antibody LeicaDS9800 (BOND™ Polymer Refine Detection).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ADA antibody [EPR25429-117] (BSA and Azide free) (ab300051), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ADA antibody [EPR25429-117] (BSA and Azide free) (ab300051)

    This data was developed using Anti-ADA antibody [EPR25429-117] ab300050, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling ADA with Anti-ADA antibody [EPR25429-117] ab300050 at 1/2000 followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Positive staining on the stromal cells of human liver. The section was incubated with Anti-ADA antibody [EPR25429-117] ab300050 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

    Secondary antibody only control: PBS was used instead of primary antibody (Anti-ADA antibody [EPR25429-117] ab300050) followed by ready to use secondary antibody LeicaDS9800 (BOND™ Polymer Refine Detection).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  • Immunocytochemistry/ Immunofluorescence - Anti-ADA antibody [EPR25429-117] (BSA and Azide free) (ab300051), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-ADA antibody [EPR25429-117] (BSA and Azide free) (ab300051)

    This data was developed using Anti-ADA antibody [EPR25429-117] ab300050, the same antibody clone in a different buffer formulation.

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeilized Jurkat (human T cell leukemia T lymphocyte) cells lebelling ADA with Anti-ADA antibody [EPR25429-117] ab300050 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic and membranous staining in the Jurkat cell line.

    Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: PBS was used instead of primary antibody followed by secondary antibody (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ADA antibody [EPR25429-117] (BSA and Azide free) (ab300051), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ADA antibody [EPR25429-117] (BSA and Azide free) (ab300051)

    This data was developed using Anti-ADA antibody [EPR25429-117] ab300050, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labeling ADA with Anti-ADA antibody [EPR25429-117] ab300050 at 1/2000 followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection) was used. Negative control: no staining on human cerebrum (PMID: 2606352). The section was incubated with Anti-ADA antibody [EPR25429-117] ab300050 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: PBS was used instead of primary antibody (Anti-ADA antibody [EPR25429-117] ab300050) followed by ready to use secondary antibody LeicaDS9800 (BOND™ Polymer Refine Detection).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ADA antibody [EPR25429-117] (BSA and Azide free) (ab300051), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ADA antibody [EPR25429-117] (BSA and Azide free) (ab300051)

    This data was developed using Anti-ADA antibody [EPR25429-117] ab300050, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded human tonsil tissue labelling ADA with Anti-ADA antibody [EPR25429-117] ab300050 at 1/2000 followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Positive staining on human tonsil (PMID: 30778076) (PMID: 6360330). The section was incubated with Anti-ADA antibody [EPR25429-117] ab300050 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: PBS was used instead of primary antibody (Anti-ADA antibody [EPR25429-117] ab300050) followed by ready to use secondary antibody LeicaDS9800 (BOND™ Polymer Refine Detection).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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