Immunofluorescent analysis of HIF1-alpha was performed on 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0. 25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with ABfinity™ HIF1-alpha recombinant rabbit monoclonal antibody (Product # 700505) at a dilution of 1:400 in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Flour® 488 goat anti-rabbit IgG secondary antibody (Product # A11008) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938) F-actin (Panel c: red) was stained with Alexa Fluor® 594 phalloidin (Product # A12381) and panel d is a merged image showing cytoplasmic localization. Panel e is a control without primary antibody. The images were captured using a Nikon microscope at 20X magnification.
|Tested species reactivity||Human|
|Published species reactivity||Not Applicable|
|Host / Isotype||Rabbit / IgG|
|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 *|
|ChIP assay (ChIP)||1 ul|
|Flow Cytometry (Flow)||1:100-1:500|
|Western Blot (WB)||1:500-1:5000|
* 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.
|Western Blot (WB)||See 1 publications below|
This antibody is predicted to react with mouse 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™ 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.
HIF1-alpha is a subunit HIF1, which is a transcription factor found in mammalian cells cultured under reduced oxygen tension. HIF1 functions as a transcriptional regulator of the adaptive response to hypoxia. HIF1-alpha regulates hypoxia-mediated apoptosis, cell proliferation and tumour angiogenesis. Hypoxia which induces p53 protein accumulation, directly interacts with HIF1-alpha and reduces hypoxia-induced expression of HIF1-alpha by promoting MDM2-mediated ubiquitination and proteasomal degradation under hypoxic conditions. Recent studies suggests that induction of NOX4 by HIF1-alpha contributes to maintain ROS levels after hypoxia and hypoxia-induced proliferation. In humans, it is located on the q arm of chromosome 14.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Thyroid Transcription Factor 1 Reprograms Angiogenic Activities of Secretome.
700505 was used in western blot to study reprogramming of angiogenic activities of secretome by thyroid transcription factor 1
|Wood LW,Cox NI,Phelps CA,Lai SC,Poddar A,Talbot C,Mu D||Scientific reports (6:null)||2016|