Western blot analysis of Rab4 was performed by loading 20 µg of HeLa (lane1), SH-SY5Y (lane2), COS-7 (lane3) and U-87 MG (lane4) cell lysates using Novex® NuPAGE® 4-12 % Bis-Tris gel (NP0321BOX), XCell SureLock™ Electrophoresis System (EI0002), Novex® Sharp Pre-Stained Protein Standard (LC5800), and iBlot® Dry Blotting System (IB21001). Proteins were transferred to a nitrocellulose membrane and blocked with 5 % skim milk for 1 hour at room temperature. Rab4 was detected at ~24 kDa using ABfinity™ Rab4 Recombinant Rabbit Oligoclonal Antibody (710543) at 1-3 µg/ml dilution in 2.5 % skim milk at 4°C overnight on a rocking platform. Goat Anti-Rabbit IgG-HRP Secondary Antibody (G21234) at 1:5000 dilution was used and chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (WP20005).
|Tested species reactivity||Human, Non-human primate|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Peptide corresponding to amino acids 193–206 of human RAB4|
|Contains||0.09% sodium azide|
|Storage Conditions||Maintain refrigerated at 2-8°C for up to 1 month. For long term storage store at -20°C|
|Tested Applications||Dilution *|
|Western Blot (WB)||1-3µg/ml|
* 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.
This antibody is predicted to react with bovine, canine, equine, feline, non-human primate, mouse, rabbit, rat and ovine based on sequence homology.
ABfinity™ recombinant antibodies are rabbit monoclonal antibodies, unmatched for producing superior results. ABfinity™ antibodies are developed by immunizing animals, screening for functionality, cloning the immunogen-specific antibody genes into high-level mammalian expression vectors, produced on a large scale and purified with Protein A.
ABfinity™ oligoclonal antibodies comprise a selection of multiple different recombinant monoclonal antibodies, providing the best of both worlds—the sensitivity of a polyclonal antibody with the specificity of a monoclonal, all delivered with the consistency only found in a recombinant antibody. While functionally the same as a polyclonal antibody—recognizing multiple epitope sites on the target and producing higher detection sensitivity for low abundance targets when compared with monoclonal antibodies—an oligoclonal antibody has a known mixture of light and heavy chains. This exact population can be produced in every lot, circumventing the biological variability typically associated with polyclonal antibody production.
Intact IgG appears on a non-reducing gel as ~150 kDa band and upon reduction generating a ~25 kDa light chain band and a ~50 kDa heavy chain.
RAB4 is a GTPase belonging to the Ras superfamily which functions in both endocytotic and exocytotic pathways, and is regulated by GTPGDP exchange cycles. In organelle membranes, RAB4 molecules are highly compartmentalized and functions as a key regulator for sorting and recycling of the membrane and proteins. RAB4 is present on all endosome populations.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.