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Adenosine Deaminase Peptide 1

Antibody: CPTC-ADA-1
Background

Catalog Number:

CPTC-ADA-1

RRID:

AB_2890075

Target Antigen:

Adenosine Deaminase Peptide 1

Isotype:

IgG

Species:

Rabbit Monoclonal Antibody

Last Updated:

07/03/2024

Antigen Recognition(s):

Peptide, Recombinant Full-length, Endogenous

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Characterization Data [Compare Characterization Data]
  Affinity Measurement by Biolayer Interferometry
Click to enlarge image Affinity and binding kinetics of CPTC-ADA-1 antibody and full length ADA recombinant protein were measured using biolayer interferometry. CPTC-ADA-1 rabbit antibody was captured onto Protein A biosensors. Recombinant protein at 1024 nM, 256 nM, 64 nM, 16 nM, and 4 nM was used as analyte. Buffer only and biosensors immobilized without antibody were used as references for background subtraction. All binding data were analyzed globally using a 1:1 fitting model. Click image to enlarge

CPTC-ADA-1 Affinity and Kinetics (Bio-Layer Interferometry) 

Result: Positive

Affinity and binding kinetics of CPTC-ADA-1 antibody and full length ADA recombinant protein were measured using biolayer interferometry. CPTC-ADA-1 rabbit antibody was captured onto Protein A biosensors. Recombinant protein at 1024 nM, 256 nM, 64 nM, 16 nM, and 4 nM was used as analyte. Buffer only and biosensors immobilized without antibody were used as references for background subtraction. All binding data were analyzed globally using a 1:1 fitting model.


Characterization SOP Files

  Affinity Measurement by SPR
Click to enlarge image Affinity and binding kinetics of CPTC-ADA-1 and full-length ADA recombinant protein were measured using surface plasmon resonance. CPTC-ADA-1 antibody was captured on a protein A biosensor chip. ADA recombinant protein was at, 256 nM, 64 nM, 16 nM, and 4 nM was used as analyte. All binding data were double referenced and analyzed globally using a 1:1 fitting model. Click image to enlarge

CPTC-ADA-1 Affinity and Kinetics (Surface Plasmon Resonance)

Result: Positive

Affinity and binding kinetics of CPTC-ADA-1 and full-length ADA recombinant protein were measured using surface plasmon resonance. CPTC-ADA-1 antibody was captured on a protein A biosensor chip. ADA recombinant protein was at, 256 nM, 64 nM, 16 nM, and 4 nM was used as analyte. All binding data were double referenced and analyzed globally using a 1:1 fitting model.


Characterization SOP Files

  IHC Tissue
Click to enlarge image Tissue Micro-Array (TMA) core of prostate cancer showing cytoplasmic staining using Antibody CPTC-ADA-1. Titer: 1:1250 Click image to enlarge

CPTC-ADA-1 IHC Tissue

Result: Positive

Tissue Micro-Array (TMA) core of prostate cancer showing cytoplasmic staining using Antibody CPTC-ADA-1. Titer: 1:1250


Characterization SOP Files

  IP

CPTC-ADA-1 Immunoprecipitation

Result: Negative

Immuno-precipitation performed using CPTC-ADA-1 as capture antibody against the over-expressed lysate of ADA (Myc-tagged). Eluates were tested in automated western blot using the an anti-Myc tag as detection antibody. The target protein was not pulled down.


Characterization SOP Files

  Immuno-MRM
Click to enlarge image Immuno-MRM chromatogram of CPTC-ADA-1 antibody (see CPTAC assay portal for details: https://assays.cancer.gov/CPTAC-6234 for oxidized peptide and https://assays.cancer.gov/CPTAC-6227 for not oxidized peptide) Data provided by the Paulovich Lab, Fred Hutch (https://research.fredhutch.org/paulovich/en.html). Data shown were obtained from frozen tissue Click image to enlarge

CPTC-ADA-1 iMRM (Frozen Tissue)

Result: Positive

Immuno-MRM chromatogram of CPTC-ADA-1 antibody (see CPTAC assay portal for details: https://assays.cancer.gov/CPTAC-6234 for oxidized peptide and https://assays.cancer.gov/CPTAC-6227 for not oxidized peptide)
Data provided by the Paulovich Lab, Fred Hutch (https://research.fredhutch.org/paulovich/en.html). Data shown were obtained from frozen tissue


Characterization SOP Files

  Immunofluorescence

CPTC-ADA-1 Immunofluorescence

Result: Negative

Immunofluorescence staining of HeLa and MCF7 cells using CPTC-ADA-1 antibody. ADA protein expression was not detected in either cell lines.


Characterization SOP Files

  NCI 60 Protein Array

CPTC-ADA-1 NCI 60 Protein Array

Result: Negative

This antibody is not suitable for use in a Reverse Phase Protein Array format as described in SOP M-105.


Characterization SOP Files

  Western Blot - Recombinant Protein or Peptide
Click to enlarge image Western blot using CPTC-ADA-1 as primary antibody against Adenosine deaminase (1-363, His-tag) human recombinant protein (lane 2). Expected molecular weight - 42.9 kDa. Molecular weight standards are also included (lane 1). Click image to enlarge

CPTC-ADA-1 Western Blot (Recombinant Protein)

Result: Positive

Western blot using CPTC-ADA-1 as primary antibody against Adenosine deaminase (1-363, His-tag) human recombinant protein (lane 2). Expected molecular weight - 42.9 kDa. Molecular weight standards are also included (lane 1).


Characterization SOP Files

  Western Blot - Tissue or Cell Lysate
Click to enlarge image Western blot using CPTC-ADA-1 as primary antibody against HeLa (lane 2), Jurkat (lane 3), A549 (lane 4), MCF7 (lane 5), and H226 (lane 6) whole cell lysates. Expected molecular weight - 40.8 kDa. Molecular weight standards are also included (lane 1). Click image to enlarge

CPTC-ADA-1 Western Blot (Cell Lysate)

Result: Positive

Western blot using CPTC-ADA-1 as primary antibody against HeLa (lane 2), Jurkat (lane 3), A549 (lane 4), MCF7 (lane 5), and H226 (lane 6) whole cell lysates. Expected molecular weight - 40.8 kDa. Molecular weight standards are also included (lane 1).


Characterization SOP Files

  Western Blot - Tissue or Cell Lysate
Click to enlarge image Automated western blot using CPTC-ADA-1 as primary antibody against buffy coat (lane 2), HeLa (lane 3), Jurkat (lane 4), A549 (lane 5), MCF7 (lane 6), and H226 (lane 7) cell lysates. Expected molecular weight - 40.8 kDa. Buffy coat and all cell lines are positive. Molecular weight standards are also included (lane 1). Click image to enlarge

CPTC-ADA-1 Automated Western Blot (Cell Lysate)

Result: Positive

Automated western blot using CPTC-ADA-1 as primary antibody against buffy coat (lane 2), HeLa (lane 3), Jurkat (lane 4), A549 (lane 5), MCF7 (lane 6), and H226 (lane 7) cell lysates. Expected molecular weight - 40.8 kDa. Buffy coat and all cell lines are positive. Molecular weight standards are also included (lane 1).


Characterization SOP Files

  Western Blot - Tissue or Cell Lysate
Click to enlarge image Western blot using CPTC-ADA-1 as primary antibody against human breast (2), ovary (3), spleen (4), endometrium (5), and lung (6) tissue lysates. The expected molecular weight is 40.7 kDa. Cytochrome C was used as a loading control. Click image to enlarge

CPTC-ADA-1 Western Blot (Tissue Lysates)

Result: Positive

Western blot using CPTC-ADA-1 as primary antibody against human breast (2), ovary (3), spleen (4), endometrium (5), and lung (6) tissue lysates. The expected molecular weight is 40.7 kDa. Cytochrome C was used as a loading control.


Characterization SOP Files

  Western Blot - Tissue or Cell Lysate
Click to enlarge image Automated WB (Simple Western) using CPTC-ADA-1 as primary antibody against the whole lysate of breast, ovary, spleen, endometrium and lung tissues. Expected MW is 40.8 Kda. The antibody presumably recognize the target in tested lysates. The same cell lysates were probed with an anti-CytC antibody (bottom panel). Click image to enlarge

CPTC-ADA-1 (tissue lysates)

Result: Presumed Positive (with additional bands)

Automated WB (Simple Western) using CPTC-ADA-1 as primary antibody against the whole lysate of breast, ovary, spleen, endometrium and lung tissues. Expected MW is 40.8 Kda. The antibody presumably recognize the target in tested lysates. The same cell lysates were probed with an anti-CytC antibody (bottom panel).


Characterization SOP Files

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Antigen: Adenosine Deaminase Peptide 1
Background

NCI Identification Number:

00344

Antigen Name:

Adenosine Deaminase Peptide 1

CPTC Name:

CPTC-ADA Peptide 1

Aliases:

Adenosine Deaminase; Adenosine Aminohydrolase; EC 3.5.4.4; ADA1; ADA

Function:

This gene encodes an enzyme that catalyzes the hydrolysis of adenosine to inosine in the purine catabolic pathway. Various mutations have been described for this gene and have been linked to human diseases related to impaired immune function such as severe combined immunodeficiency disease (SCID) which is the result of a deficiency in the ADA enzyme. In ADA-deficient individuals there is a marked depletion of T, B, and NK lymphocytes, and consequently, a lack of both humoral and cellular immunity. Conversely, elevated levels of this enzyme are associated with congenital hemolytic anemia. ADA (Adenosine Deaminase) is a Protein Coding gene. Diseases associated with ADA include Severe Combined Immunodeficiency, Autosomal Recessive, T Cell-Negative, B Cell-Negative, Nk Cell-Negative, Due To Adenosine Deaminase Deficiency and Severe Combined Immunodeficiency. Among its related pathways are Metabolism of nucleotides and C-MYB transcription factor network. Gene Ontology (GO) annotations related to this gene include adenosine deaminase activity and purine nucleoside binding. An important paralog of this gene is ADAL.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 (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). ADA_HUMAN,P00813Deaminases are enzymes that catalyze the hydrolysis of C-NH2 bonds in amino compounds, producing ammonia. They are usually named after their substrate; two well known members of this group are adenosine deaminase and cytidine deaminase.

Chromosomal Localization:

20q13.12

Accession Number:

NP_000013.2

UniProt Accession Number:

P00813

DNA Source:

N/A

Immunogen:

Synthetic Peptide

Vector Name:

N/A

Extinction Coefficient:

Buffers:

Expressed Sequence:

STLDTDYQMTK

Native Sequence:

Calculated Isoelectric Point:

Molecular Weight:

1210

Last Updated:

09/02/2020

Links

Characterization Data

SOPs

No SOPs available.

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