Antibodies, Proteins, Kits and Reagents for Life Science

321 638 ₸ Product size

100 µl 321 638 ₸

Order now and get it on Tuesday March 02, 2021

Anti-DDAH1 antibody [EPR13922] (ab180599)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR13922] to DDAH1
Suitable for: Flow Cyt, WB, IHC-P, ICC/IF, IP
Knockout validated
Reacts with: Human

Product name

Anti-DDAH1 antibody [EPR13922]
See all DDAH1 primary antibodies
Description

Rabbit monoclonal [EPR13922] to DDAH1
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC/IF	Human
IHC-P	Human
IP	Human
WB	Human

See all applications and species data

Immunogen

This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: HuVEC, A431 and HepG2 Human fetal brain, liver, kidney or 293 lysate, Human ovarian carcinoma tissue, 293 cells.
General notes
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 production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant
Concentration information loading...
Purity

Tissue culture supernatant
Clonality

Monoclonal
Clone number

EPR13922
Isotype

IgG
Research areas

Western blot - Anti-DDAH1 antibody [EPR13922] (ab180599)

All lanes : Anti-DDAH1 antibody [EPR13922] (ab180599) at 1/10000 dilution

Lane 1 : A431 wild-type cell lysate
Lane 2 : A431 DDAH1 knockout cell lysate
Lane 3 : HuVEC cell lysate
Lane 4 : HepG2 cell lysate

Lysates/proteins at 40 µg per lane.

Performed under reducing conditions.

Observed band size: 37 kDa
why is the actual band size different from the predicted?

Lanes 1 - 4: Merged signal (red and green). Green - ab180599 observed at 37 kDa. Red - loading control, ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.

ab180599 was shown to react with DDAH1 in A431 wild-type cells in Western blot. Loss of signal was observed when DDAH1 knockout sample was used. A431 wild-type and DDAH1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% Milk in TBS-T (0.1% Tween®) before incubation with ab180599 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4°C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were developed 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.
Western blot - Anti-DDAH1 antibody [EPR13922] (ab180599)

All lanes : Anti-DDAH1 antibody [EPR13922] (ab180599) at 1/20000 dilution

Lane 1 : Human fetal brain lysate at 20 µg
Lane 2 : Human fetal liver lysate at 20 µg
Lane 3 : Human fetal kidney lysate at 20 µg
Lane 4 : 293 lysate at 10 µg

Secondary
All lanes : Goat Anti-Rabbit IgG (H+L) Peroxidase conjugated at 1/1000 dilution

Observed band size: 37 kDa why is the actual band size different from the predicted?
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DDAH1 antibody [EPR13922] (ab180599)

Immunohistochemical analysis of paraffin embedded Human ovarian carcinoma tissue labeling DDAH1 with ab180599 at a 1/50 dilution. Prediluted HRP Polymer for Rabbit IgG secondary used. Counterstained with Hematoxylin.

Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
Flow Cytometry - Anti-DDAH1 antibody [EPR13922] (ab180599)

ab180599 staining DDAH1 in the human cell line 293 (human embryonic kidney) by flow cytometry. Cells were fixed with 4% paraformaldehyde, permabilised with 90% methanol and the sample was incubated with the primary antibody at a dilution of 1/20. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody.

Isoytype control: Rabbit monoclonal IgG (Black)

Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)
Immunocytochemistry/ Immunofluorescence - Anti-DDAH1 antibody [EPR13922] (ab180599)

Immunofluorescence analysis of 293 cells (fixative 4% paraformaldehyde) labeling DDAH1 with ab180599 at a 1/100 dilution (left image), and counterstained with Dapi (right image). Goat anti rabbit IgG (Dylight 555) secondary used at a 1/200 diution.
Immunoprecipitation - Anti-DDAH1 antibody [EPR13922] (ab180599)

Immunoprecipitation analysis of Human fetal kidney lysate labeling DDAH1 with ab180599 at a 1/40 dilution. Goat Anti-Rabbit IgG (H+L) Peroxidase conjugated secondary used at a 1/1000 dilution.
Anti-DDAH1 antibody [EPR13922] (ab180599)

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