Anti-DDAH1 antibody (ab108088)
Key features and details
- Rabbit polyclonal to DDAH1
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-DDAH1 antibody
See all DDAH1 primary antibodies -
Description
Rabbit polyclonal to DDAH1 -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species IHC-P HumanWB Human
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Immunogen
Synthetic peptide conjugated to KLH derived from within residues 200 to the C-terminus of Human DDAH1.
Read Abcam's proprietary immunogen policy -
Positive control
- WB: Wild-type A431 cell lysate. A431, HUVEC, HepG2 and HEK293 cell lysates; Human fetal liver, liver, kidney, brain and pancreas cell lystates. IHC-P: Human kidney tissue.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.4
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab108088 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
GuaranteedTested applications are guaranteed to work and covered by our Abpromise guarantee.
PredictedPredicted to work for this combination of applications and species but not guaranteed.
IncompatibleDoes not work for this combination of applications and species.
Application Species IHC-P HumanWB HumanAll applications MouseRatHorseCowDogPigMacaque monkeyChinese hamsterApplication Abreviews Notes WB Use a concentration of 1 µg/ml. Detects a band of approximately 37 kDa (predicted molecular weight: 31 kDa).IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.Notes WB
Use a concentration of 1 µg/ml. Detects a band of approximately 37 kDa (predicted molecular weight: 31 kDa).IHC-P
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.Target
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Function
Hydrolyzes N(G),N(G)-dimethyl-L-arginine (ADMA) and N(G)-monomethyl-L-arginine (MMA) which act as inhibitors of NOS. Has therefore a role in the regulation of nitric oxide generation. -
Tissue specificity
Detected in brain, liver, kidney and pancreas, and at low levels in skeletal muscle. -
Sequence similarities
Belongs to the DDAH family. - Information by UniProt
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Database links
- Entrez Gene: 23576 Human
- Entrez Gene: 69219 Mouse
- Entrez Gene: 64157 Rat
- Omim: 604743 Human
- SwissProt: P56965 Cow
- SwissProt: O94760 Human
- SwissProt: Q9CWS0 Mouse
- SwissProt: O08557 Rat
see all -
Alternative names
- DDAH antibody
- DDAH I antibody
- DDAH-1 antibody
see all
Images
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Western blot - Anti-DDAH1 antibody (ab108088)All lanes : Anti-DDAH1 antibody (ab108088) at 1 µg/ml
Lane 1 : Wild-type A431 cell lysate
Lane 2 : A431 knockout DDAH1 cell lysate
Lane 3 : HuVEC cell lysate
Lane 4 : HepG2 cell lysate
Lysates/proteins at 40 µg per lane.
Performed under reducing conditions.
Predicted band size: 31 kDa
Observed band size: 35 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab108088 observed at 35 kDa. Red - loading control, ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.
ab108088 was shown to react with DDAH1 in wild-type A431 cells in western blot. Loss of signal was observed when DDAH1 knockout sample was used. Wild-type and DDAH1 knockout A431 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab108088 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4°C at 1 µg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DDAH1 antibody (ab108088)IHC image of DDAH1 staining in human kidney formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab108088, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times. -
Western blot - Anti-DDAH1 antibody (ab108088)All lanes : Anti-DDAH1 antibody (ab108088) at 1 µg/ml
Lane 1 : Lung (Human) Whole Cell Lysate - fetal normal tissue (ab30282)
Lane 2 : Human liver tissue lysate - total protein (ab29889)
Lane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lane 4 : Human kidney tissue lysate - total protein (ab30203)
Lane 5 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lane 6 : Human brain tissue lysate - total protein (ab29466)
Lane 7 : Human pancreas tissue lysate - total protein (ab29816)
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 31 kDa
Observed band size: 37 kDa why is the actual band size different from the predicted?
Additional bands at: 62 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 1 minute
Protocols
Datasheets and documents
References (1)
ab108088 has been referenced in 1 publication.
- Osbun JW et al. Comparative Proteomic Profiling Using Two-Dimensional Gel Electrophoresis and Identification via LC-MS/MS Reveals Novel Protein Biomarkers to Identify Aggressive Subtypes of WHO Grade I Meningioma. J Neurol Surg B Skull Base 78:371-379 (2017). PubMed: 28875114
Images
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Western blot - Anti-DDAH1 antibody (ab108088)All lanes : Anti-DDAH1 antibody (ab108088) at 1 µg/ml
Lane 1 : Wild-type A431 cell lysate
Lane 2 : A431 knockout DDAH1 cell lysate
Lane 3 : HuVEC cell lysate
Lane 4 : HepG2 cell lysate
Lysates/proteins at 40 µg per lane.
Performed under reducing conditions.
Predicted band size: 31 kDa
Observed band size: 35 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab108088 observed at 35 kDa. Red - loading control, ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.
ab108088 was shown to react with DDAH1 in wild-type A431 cells in western blot. Loss of signal was observed when DDAH1 knockout sample was used. Wild-type and DDAH1 knockout A431 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab108088 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4°C at 1 µg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DDAH1 antibody (ab108088)IHC image of DDAH1 staining in human kidney formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab108088, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times. -
Western blot - Anti-DDAH1 antibody (ab108088)All lanes : Anti-DDAH1 antibody (ab108088) at 1 µg/ml
Lane 1 : Lung (Human) Whole Cell Lysate - fetal normal tissue (ab30282)
Lane 2 : Human liver tissue lysate - total protein (ab29889)
Lane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lane 4 : Human kidney tissue lysate - total protein (ab30203)
Lane 5 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lane 6 : Human brain tissue lysate - total protein (ab29466)
Lane 7 : Human pancreas tissue lysate - total protein (ab29816)
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 31 kDa
Observed band size: 37 kDa why is the actual band size different from the predicted?
Additional bands at: 62 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 1 minute
