Western blot analysis of GATA1 was performed by loading 20 µg of K562 cell lysate 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. GATA1 was detected at ~ 50 kDa using ABfinity™GATA1 Recombinant Rabbit Monoclonal Antibody (700727) at 1-3 µg/mL 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|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Recombinant protein corresponding to amino acids 1-200 of human GATA1.|
|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)||1-3 ug/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.
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™ monoclonal antibodies resemble rabbit monoclonals isolated from serum or produced by hybridomas, but demonstrate greater specificity and sensitivity. Because ABfinity™ recombinant antibodies are derived from cloned DNA sequences of the heavy and light antibody chains, they are not susceptible to cell-line drift or lot-to-lot variation, thus allowing for peak specificity and performance.
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.
Globin transcription factor (GATA) is a family of proteins that play a very important role in translating various signaling pathways which control the erythroid progenitor cells; to proliferate and differentiate. There are 6 transcription factors in the GATA family from GATA1 to GATA 6 of which GATA 1 plays a key role in the blood cell differentiation. To facilitate differentiation of red blood cells; GATA 1 along with binding to DNA also interacts with other regulatory proteins. GATA proteins have a high affinity to bind to DNA that show the [AT]GATA[AG] sequence. GATA 1 is found on the X chromosome.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.