Anti-Zyxin antibody (ab71842)
Key features and details
- Rabbit polyclonal to Zyxin
- Suitable for: IP, ICC/IF, WB
- Reacts with: Rat, Human
- Isotype: IgG
Overview
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Product name
Anti-Zyxin antibody
See all Zyxin primary antibodies -
Description
Rabbit polyclonal to Zyxin -
Host species
Rabbit -
Tested applications
Suitable for: IP, ICC/IF, WBmore details -
Species reactivity
Reacts with: Rat, Human
Predicted to work with: Mouse, Dog, Rhesus monkey
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Immunogen
Synthetic peptide corresponding to Human Zyxin aa 450-550 conjugated to keyhole limpet haemocyanin.
(Peptide available asab71841) -
Positive control
- Purchase matching WB positive control:Recombinant Human Zyxin protein
- This antibody gave a positive signal in the following Lysates: SK N SH Whole Cell, HeLa Whole Cell, A498 Whole Cell, U-87 MG Whole Cell, DU 145 Whole Cell, Rat Testis Tissue, HCT 116 Whole Cell (data not shown)
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General notes
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
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: 7.40
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
Our Abpromise guarantee covers the use of ab71842 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 IP Use a concentration of 5 µg/ml. ICC/IF Use a concentration of 5 µg/ml. WB Use a concentration of 1 µg/ml. Detects a band of approximately 75 kDa (predicted molecular weight: 61 kDa). Target
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Relevance
Zyxin binds alpha-actinin and the CRP proteins and is implicated in the regulation and modulation of actin filaments organization, cell adhesion, cell motility, mitosis, and signal transduction. Zyxin may shuttle between focal adhesion plaques and the nucleus. It has been suggested that zyxin is involved in gene transcription by its association with transcription factors in the nucleus. -
Cellular localization
Cytoplasmic, cytoskeletal and Nuclear. -
Database links
- Entrez Gene: 7791 Human
- Entrez Gene: 22793 Mouse
- Entrez Gene: 114636 Rat
- Entrez Gene: 706407 Rhesus monkey
- Omim: 602002 Human
- SwissProt: Q15942 Human
- SwissProt: Q62523 Mouse
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Alternative names
- ESP 2 antibody
- ESP2 antibody
- HED 2 antibody
see all
Images
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Western blot - Anti-Zyxin antibody (ab71842)All lanes : Anti-Zyxin antibody (ab71842) at 1 µg/ml
Lane 1 : SK N SH (Human neuroblastoma) Whole Cell Lysate
Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 3 : A498 (Human Kidney Carcinoma) Whole Cell Lysate
Lane 4 : U-87 MG (Human glioblastoma astrocytoma) Whole Cell Lysate
Lane 5 : DU 145 (Human prostate carcinoma cell line) Whole Cell Lysate
Lane 6 : Testis (Rat) Tissue Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size: 61 kDa
Observed band size: 75 kDa why is the actual band size different from the predicted?
Additional bands at: 100 kDa, 48 kDa. We are unsure as to the identity of these extra bands. -
Immunoprecipitation - Anti-Zyxin antibody (ab71842)Zyxin was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to Zyxin and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab71842.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 75kDa; Zyxin -
Immunocytochemistry/ Immunofluorescence - Anti-Zyxin antibody (ab71842)ICC/IF image of ab71842 stained HepG2 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal Goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab71842, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) HeLa cells at 5µg/ml, and in 100% Methanol fixed (5 min) Hela, Hek293, HepG2, and MCF-7 cells at 5µg/ml.
Protocols
Datasheets and documents
References (9)
ab71842 has been referenced in 9 publications.
- Tonazzini I et al. Neuronal contact guidance and YAP signaling on ultra-small nanogratings. Sci Rep 10:3742 (2020). PubMed: 32111918
- Tonazzini I et al. The role of ubiquitin ligase E3A in polarized contact guidance and rescue strategies in UBE3A-deficient hippocampal neurons. Mol Autism 10:41 (2019). PubMed: 31798818
- Sikorski K et al. A high-throughput pipeline for validation of antibodies. Nat Methods 15:909-912 (2018). PubMed: 30377371
- Albecka A et al. Dual Function of the pUL7-pUL51 Tegument Protein Complex in Herpes Simplex Virus 1 Infection. J Virol 91:N/A (2017). ICC ; Human . PubMed: 27852850
- Harrington KM & Clevenger CV Identification of NEK3 Kinase Threonine 165 as a Novel Regulatory Phosphorylation Site That Modulates Focal Adhesion Remodeling Necessary for Breast Cancer Cell Migration. J Biol Chem 291:21388-21406 (2016). PubMed: 27489110
- Van Itallie CM et al. Biotin ligase tagging identifies proteins proximal to E-cadherin, including lipoma preferred partner, a regulator of epithelial cell-cell and cell-substrate adhesion. J Cell Sci 127:885-95 (2014). PubMed: 24338363
- Ketscher A et al. LSD1 controls metastasis of androgen-independent prostate cancer cells through PXN and LPAR6. Oncogenesis 3:e120 (2014). WB, IP ; Human . PubMed: 25285406
- Vyas S et al. A systematic analysis of the PARP protein family identifies new functions critical for cell physiology. Nat Commun 4:2240 (2013). PubMed: 23917125
- Watanabe-Nakayama T et al. Requirement of LIM domains for the transient accumulation of paxillin at damaged stress fibres. Biol Open 2:667-74 (2013). ICC/IF . PubMed: 23862014
Images
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Western blot - Anti-Zyxin antibody (ab71842)All lanes : Anti-Zyxin antibody (ab71842) at 1 µg/ml
Lane 1 : SK N SH (Human neuroblastoma) Whole Cell Lysate
Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 3 : A498 (Human Kidney Carcinoma) Whole Cell Lysate
Lane 4 : U-87 MG (Human glioblastoma astrocytoma) Whole Cell Lysate
Lane 5 : DU 145 (Human prostate carcinoma cell line) Whole Cell Lysate
Lane 6 : Testis (Rat) Tissue Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size: 61 kDa
Observed band size: 75 kDa why is the actual band size different from the predicted?
Additional bands at: 100 kDa, 48 kDa. We are unsure as to the identity of these extra bands.
-
Immunoprecipitation - Anti-Zyxin antibody (ab71842)Zyxin was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to Zyxin and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab71842.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 75kDa; Zyxin -
Immunocytochemistry/ Immunofluorescence - Anti-Zyxin antibody (ab71842)ICC/IF image of ab71842 stained HepG2 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal Goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab71842, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) HeLa cells at 5µg/ml, and in 100% Methanol fixed (5 min) Hela, Hek293, HepG2, and MCF-7 cells at 5µg/ml.
