Recombinant Anti-DUSP4 antibody [EPR19881] - BSA and Azide free (ab222487)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR19881] to DUSP4 - BSA and Azide free
- Suitable for: Flow Cyt (Intra), ICC/IF, WB, IP
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
-
Product name
Anti-DUSP4 antibody [EPR19881] - BSA and Azide free
See all DUSP4 primary antibodies -
Description
Rabbit monoclonal [EPR19881] to DUSP4 - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), ICC/IF, WB, IPmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.
-
Positive control
- WB: MDA-MB-231, A549, Wild-type A549, SK-BR-3, HCT 116, RAW 264.7, PC-12, MOLT-4 and C6 whole cell lysates; Human breast cancer lysate. ICC/IF: A549 and MDA-MB-231 cells. Flow Cyt (intra): MDA-MB-231 cells, A549 cells. IP: MDA-MB-231 whole cell lysate.
-
General notes
ab222487 is the carrier-free version of ab216576.
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.
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 production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
-
Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
-
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR19881 -
Isotype
IgG -
Research areas
Associated products
-
Alternative Versions
-
Compatible Secondaries
-
Conjugation kits
-
Isotype control
-
Recombinant Protein
-
Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab222487 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
---|---|---|
Flow Cyt (Intra) |
Use at an assay dependent concentration.
|
|
ICC/IF |
Use at an assay dependent concentration.
This product gave a positive signal in A549 (DUSP4 knockout A549 cells used as a negative control) fixed with 100% methanol (5 min). |
|
WB |
Use at an assay dependent concentration. Detects a band of approximately 43 kDa (predicted molecular weight: 43 kDa).
|
|
IP |
Use at an assay dependent concentration.
|
Notes |
---|
Flow Cyt (Intra)
Use at an assay dependent concentration. |
ICC/IF
Use at an assay dependent concentration. This product gave a positive signal in A549 (DUSP4 knockout A549 cells used as a negative control) fixed with 100% methanol (5 min). |
WB
Use at an assay dependent concentration. Detects a band of approximately 43 kDa (predicted molecular weight: 43 kDa). |
IP
Use at an assay dependent concentration. |
Target
-
Function
Regulates mitogenic signal transduction by dephosphorylating both Thr and Tyr residues on MAP kinases ERK1 and ERK2. -
Sequence similarities
Belongs to the protein-tyrosine phosphatase family. Non-receptor class dual specificity subfamily.
Contains 1 rhodanese domain.
Contains 1 tyrosine-protein phosphatase domain. -
Post-translational
modificationsPhosphorylation in the C-terminus by ERK1/2 inhibits proteasomal degradation and stabilizes the protein. -
Cellular localization
Nucleus. - Information by UniProt
-
Database links
- Entrez Gene: 1846 Human
- Entrez Gene: 319520 Mouse
- Entrez Gene: 60587 Rat
- Omim: 602747 Human
- SwissProt: Q13115 Human
- SwissProt: Q8BFV3 Mouse
- SwissProt: Q62767 Rat
- Unigene: 417962 Human
see all -
Alternative names
- Dual specificity phosphatase 4 antibody
- Dual specificity protein phosphatase 4 antibody
- Dual specificity protein phosphatase hVH2 antibody
see all
Images
-
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab216576).
Flow cytometry overlay histogram showing left wild-type A549 positive cells and right negative DUSP4 knockout A549 stained with ab216576 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab216576) (1x 106 in 100μl at 1.0 μg/ml (1/1990)) for 30min at 22°C.
The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 22°C
Isotype control antibody Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (black line) was used at the same concentration and conditions as the primary antibody. Unlabelled sample was also used as a control (blue line).
Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.
-
All lanes : Anti-DUSP4 antibody [EPR19881] (ab216576) at 1/1000 dilution
Lane 1 : Wild-type A549 cell lysate
Lane 2 : DUSP4 knockout A549 cell lysate
Lane 3 : A549 cell lysate
Lane 4 : MOLT-4 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 43 kDa
Observed band size: 40 kDa why is the actual band size different from the predicted?This data was developed using the same antibody clone in a different buffer formulation (ab216576).
Lanes 1 - 4: Merged signal (red and green). Green - ab216576 observed at 40 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.
ab216576 was shown to react with DUSP4 in wild-type A549 cells in Western blot with loss of signal observed in DUSP4 knockout cell line ab273859 (DUSP4 knockout cell lysate ab273813). Wild-type A549 and DUSP4 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab216576 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 1000 dilution 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 h at room temperature before imaging.
-
ab216576 staining DUSP4 in wild-type A549 cells, with negative expression in DUSP4 knockout A549 cells. The cells were fixed with 100% methanol (5 min), permeabilised with 0.1% Triton x-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab216576 at 1 μg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin at 0.5 μg/ml. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150119, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 647), pre-adsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab216576).
-
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A549 (Human lung carcinoma cell line) cells labeling DUSP4 with ab216576 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing nuclear staining on A549 cell line.
The nuclear counterstain is DAPI (blue). Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab216576).
-
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MDA-MB-231 (Human breast adenocarcinoma cell line) cells labeling DUSP4 with ab216576 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing nuclear staining on MDA-MB-231 cell line.
The nuclear counterstain is DAPI (blue). Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab216576).
-
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed MDA-MB-231 (Human breast adenocarcinoma cell line) cells labeling DUSP4 with ab216576 at 1/60 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab216576).
-
DUSP4 was immunoprecipitated from 0.35mg of MDA-MB-231 (Human breast adenocarcinoma cell line) whole cell lysate with ab216576 at 1/30 dilution.
Western blot was performed from the immunoprecipitate using ab216576 at 1/500 dilution.
VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.
Lane 1: MDA-MB-231 whole cell lysate, 10µg (Input).
Lane 2: ab216576 IP in MDA-MB-231 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab216576 in MDA-MB-231 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 1 second.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab216576).
Protocols
Datasheets and documents
-
Datasheet download
Certificate of Compliance
References (0)
ab222487 has not yet been referenced specifically in any publications.