Western blot analysis was performed using membrane enriched extracts (30 µg) of Raji (Lane 1), U-87 MG (Lane 2), Hep G2 (Lane 3), Mouse Liver (Lane 4) and HeLa (Lane 5).The blots were probed with Anti-Anthrax Toxin Receptor Rabbit Polyclonal Antibody (Product # PA1-602, 1 in 1000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4µg/mL 1:2500 dilution). A~ 65kDa band of Anthrax Toxin Receptor was observed across the cell lines tested. A ~58kDa band was observed in the Mouse liver tissue sample. A band of 20 kDa was also observed across the panel of cell lines and tissues tested. This could be due to the separation of the PA subunit, of the Anthrax Toxin Receptor, into two subunits of 65kDa and 20kDa, upon trypsinolysis. A Known quantity of protein samples were electrophoresed using Novex® NuPAGE®12 % Bis-Tris gel (Product # NP0342BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane by iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
|Tested species reactivity||Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic Peptide: L(92) M K L T E D R E Q I R Q G L E(107) C|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunohistochemistry (Paraffin) (IHC (P))||1:100|
|Western Blot (WB)||1:1000|
* 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.
PA1-602 detects Tem-8 from mouse samples.
PA1-602 has been successfully used in Western blot and immunohistochemical procedures. By Western blot, this antibody detects ~45 kDa, 60 kDa, 85 kDa proteins, representing Tem-8 from mouse tissue extracts.This antibody has also been used in immunohistochemical procedures (paraffin samples) using mouse tissues.
PA1-602 immunizing peptide corresponds to amino acid residues 92-107, within the extracellular domain, of human ATR/TEM8. This peptide (Cat. # PEP-253) is available for neutralization and control experiments.
Anthrax Toxin Receptor (ATR), also known as Tem-8 is a single transmembrane domain containing protein which is alternately spliced for a short, medium and long form. The short form represents the secreted form of the protein. The mid-length form is also known as ATR (Anthrax Toxin Receptor) has known functions in the response to the lethal toxin from the anthrax bacteria. The long form is termed Tem-8 and has been shown to be involved in angiogenesis and the proteasomal degradation pathway.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.