• Species Reactivity

  Human

• Specificity

  Detects human Afadin/AF-6 in direct ELISAs.

• Source

  Monoclonal Mouse IgG    ₁ Clone # 851204

• Purification

  Protein A or G purified from hybridoma culture supernatant

• Immunogen

  E. coli-derived recombinant human Afadin/AF-6    
  Asn217-Pro392    
  Accession # P55196

• Formulation

  Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 μm filtered solution in PBS.

• Label

  Unconjugated

Applications

• 
  

  Recommended    
  Concentration

  Sample

• Western Blot

  0.5 μg/mL

  See below

• 
  

• Immunocytochemistry

  8-25 μg/mL

  See below

• 
  

Please Note: Optimal dilutions should be determined by each laboratory for each application.  are available in the Technical Information section on our website.

Data Examples

Preparation and Storage

• Reconstitution

  Reconstitute at 0.5 mg/mL in sterile PBS.

• Shipping

  The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C

• Stability & Storage

  Use a manual defrost freezer and avoid repeated freeze-thaw cycles.    

• 12 months from date of receipt, -20 to -70 °C as supplied.

• 1 month, 2 to 8 °C under sterile conditions after reconstitution.

• 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: Afadin/AF-6

Afadin, also known as MLLT-4 (Myeloid/Lymphoid or mixed-lineage Leukemia Translocated to Ch 4) and AF-6,  is a 200-210 kDa nectin and F-actin-binding intracellular protein. It is ubiquitously expressed, and serves as an intermediary that links nectin with the actin cytoskeleton in tight and adherens junctions (AJs). Afadin is involved in the nectin-mediated recruitment of catenins to zonule adherens complexes, and in the recruitment of ZO-1 to the apical side of AJs for the purpose of forming tight junctions. Human Afadin is 1824 amino acids (aa) in length. The protein contains two Ras-association (RA) domains (aa 20-348), a phosphoprotein FHA domain (aa 426?492), a Vav2 Ras-activating "dilute" domain (aa 668-908), one PDZ domain that binds protein C-termini (aa 1007-1093), and three proline-rich regions (aa 1346?1708). There are at least fourteen utilized Ser/Thr phosphorylation sites and two Tyr phosphorylation sites. There are at least seven alternative splice variants are reported for Afadin. There is a 190 kDa isoform that shows a 12 aa insert after Arg1605 coupled to a four aa substitution for aa 1650-1824. Other isoforms contain scattered deletions of one to three aa coupled to major aa changes. Five isoforms share a deletion of aa 393-407, with four of these containing a 24 aa substitution for aa 1605-1824, a 73 aa substitution for aa 1747-1824, an 11 aa substitution for aa 680-1824, and a deletion of aa 1683-1746, respectively. A final isoform possesses a four aa substitution for aa 1650-1824. Over aa 217-392, human Afadin shares 94% aa sequence identity with mouse Afadin.

• Entrez Gene IDs:

  4301 (Human); 17356 (Mouse); 26955 (Rat)

• Alternate Names:

  AF6; AF-6; AF64; AFAD; Afadin; FLJ34371; MLLT4; myeloid/lymphoid or mixed-lineage leukemia (trithorax homolog, Drosophila);translocated to, 4