Recombinant Anti-N WASP antibody [EPR6959] (ab126626)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR6959] to N WASP
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Mouse, Human
Related conjugates and formulations
Overview
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Product name
Anti-N WASP antibody [EPR6959]
See all N WASP primary antibodies -
Description
Rabbit monoclonal [EPR6959] to N WASP -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Human
Predicted to work with: Pig -
Immunogen
Synthetic peptide within Human N WASP aa 300-450. The exact sequence is proprietary.
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Positive control
- NIH 3T3, F9, L929, HeLa and K562 cell lysates; Human papillary carcinoma tissue.
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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
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
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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. Stable for 12 months at -20°C. -
Dissociation constant (KD)
KD = 7.40 x 10 -11 M Learn more about KD -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR6959 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- Alexa Fluor® 488 Anti-N WASP antibody [EPR6959] (ab208521)
- Alexa Fluor® 594 Anti-N WASP antibody [EPR6959] (ab208522)
- Alexa Fluor® 555 Anti-N WASP antibody [EPR6959] (ab208523)
- Alexa Fluor® 647 Anti-N WASP antibody [EPR6959] (ab208619)
- Anti-N WASP antibody [EPR6959] - BSA and Azide free (ab232457)
- Alexa Fluor® 568 Anti-N WASP antibody [EPR6959] (ab312455)
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Conjugation kits
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab126626 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 |
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
Purified format |
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WB | (1) |
1/1000 - 1/10000. Detects a band of approximately 65 kDa (predicted molecular weight: 55 kDa).
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IHC-P | (1) |
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
|
ICC/IF |
1/300.
For unpurified, use 1/50 - 1/100. |
Notes |
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Flow Cyt (Intra)
Use at an assay dependent concentration. Purified format |
WB
1/1000 - 1/10000. Detects a band of approximately 65 kDa (predicted molecular weight: 55 kDa). |
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
ICC/IF
1/300. For unpurified, use 1/50 - 1/100. |
Target
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Function
Regulates actin polymerization by stimulating the actin-nucleating activity of the Arp2/3 complex. Binds to HSF1/HSTF1 and forms a complex on heat shock promoter elements (HSE) that negatively regulates HSP90 expression. -
Sequence similarities
Contains 1 CRIB domain.
Contains 1 WH1 domain.
Contains 2 WH2 domains. -
Cellular localization
Cytoplasm > cytoskeleton. Nucleus. Preferentially localized in the cytoplasm when phosphorylated and in the nucleus when unphosphorylated. - Information by UniProt
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Database links
- Entrez Gene: 8976 Human
- Entrez Gene: 73178 Mouse
- Omim: 605056 Human
- SwissProt: O00401 Human
- SwissProt: Q91YD9 Mouse
- Unigene: 143728 Human
- Unigene: 83916 Human
- Unigene: 1574 Mouse
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Alternative names
- DKFZp779G0847 antibody
- MGC48327 antibody
- N-WASP antibody
see all
Images
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Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: N WASP knockout HAP1 cell lysate (20 µg)
Lane 3: A549 cell lysate (20 µg)
Lane 4: Human thyroid tissue lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab126626 observed at 67 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab126626 was shown to recognize N WASP when N WASP knockout samples were used, along with additional cross-reactive bands. Wild-type and N WASP knockout samples were subjected to SDS-PAGE. ab126626 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/2000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging. -
Immunofluorescence staining of K562 cells with purified ab126626 at a working dilution of 1/300, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab126626 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.
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Intracellular Flow Cytometry analysis of K562 (human chronic myelogenous leukemia) cells labeling N WASP with purified ab126626 at 1/150 (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat Anti-Rabbit IgG H&L (Alexa Fluorr® 488) (ab150077)was used as the secondary antibody. Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.
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Immunohistochemical staining of paraffin embedded human kidney with purified ab126626 at a working dilution of 1/100. The secondary antibody used is ab97051, a goat anti-rabbit IgG (H&L) at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
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Anti-N WASP antibody [EPR6959] (ab126626) at 1/10000 dilution (purified) + L929 whole cell lysate at 10 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 55 kDa
Observed band size: 65 kDa why is the actual band size different from the predicted? -
All lanes : Anti-N WASP antibody [EPR6959] (ab126626) at 1/10000 dilution (purified)
Lane 1 : HeLa whole cell lysate
Lane 2 : K562 whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 55 kDa
Observed band size: 65 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST -
All lanes : Anti-N WASP antibody [EPR6959] (ab126626) at 1/1000 dilution (Unpurified)
Lane 1 : NIH 3T3 cell lysate
Lane 2 : F9 cell lysate
Lane 3 : L929 cell lysate
Lane 4 : HeLa cell lysate
Lane 5 : K562 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat anti-rabbit HRP conjugated antibody at 1/2000 dilution
Predicted band size: 55 kDa -
Unpurified ab126626, at 1/50, staining N WASP in formalin fixed, paraffin embedded human papillary carcinoma tissue by Immunohistochemistry
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (13)
ab126626 has been referenced in 13 publications.
- Kaplan C et al. Load adaptation by endocytic actin networks. Mol Biol Cell 33:ar50 (2022). PubMed: 35389747
- Zou YJ et al. RAB14 GTPase is essential for actin-based asymmetric division during mouse oocyte maturation. Cell Prolif 54:e13104 (2021). PubMed: 34323331
- Hou J et al. Identify Function of WASL in Prognosis of Cervical Cancer Based on Omics Data. Front Cell Dev Biol 9:670890 (2021). PubMed: 34222242
- Zhang Y et al. RAB35 depletion affects spindle formation and actin-based spindle migration in mouse oocyte meiosis. Mol Hum Reprod 25:359-372 (2019). PubMed: 31152166
- Zhang Y et al. Loss of PKC mu function induces cytoskeletal defects in mouse oocyte meiosis. J Cell Physiol N/A:N/A (2019). PubMed: 30912144