Immunofluorescence analysis of Ephrin A4 was performed using 70% confluent log phase Jurkat cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Ephrin A4 Rabbit Polyclonal Antibody (343700) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing membranous localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse|
|Published species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide derived from the C-terminal end of the mouse ephrin-A4 protein|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunohistochemistry (IHC)||Assay Dependent|
|Immunoprecipitation (IP)||Assay Dependent|
|Western Blot (WB)||Assay Dependent|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
Ephrin A4 (the antigen to this antibody) is also known as LERK-4 or ELF-4. Ephrin-A4 binds EphA2, EphA3, EphA4, EphA5, EphA6, EphA7, and EphB1. To date, at least 14 members of the Eph receptor family and a family of 8 ligands have been identified. Ligands of Eph family receptors are structurally related membrane-bound proteins that can be subdivided into two major subclasses: ephrin-A and ephrin-B. Ligands in the ephrin-A subclass, including the prototype family member ephrin-A1 (B61), are membrane associated through glycosylphosphatidyl-inositollinkages; whereas ephrin-B subclass consists of ligands with transmembrane domains.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Evaluation of Eph receptor and ephrin expression within the human cornea and limbus.
34-3700 was used in immunohistochemistry to study Eph/ephrin expression within the human cornea.
|Hogerheyde TA,Stephenson SA,Harkin DG,Bray LJ,Madden PW,Woolf MI,Richardson NA||Experimental eye research (107:110)||2013|
|Human||Not Cited||Genome-wide analysis of promoter methylation associated with gene expression profile in pancreatic adenocarcinoma.||Vincent A,Omura N,Hong SM,Jaffe A,Eshleman J,Goggins M||Clinical cancer research : an official journal of the American Association for Cancer Research (17:4341)||2011|