Immunofluorescent analysis of SREBP2 (green) showing staining in the cytoplasm of HeLa cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a SREBP2 polyclonal antibody (Product # PA1-338) in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4 °C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a flourescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues A(843) I S W L Q G D D A A V R S H(857) of rat SREBP 2.|
|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 *|
|Western Blot (WB)||1:100-1:1000|
* 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.
|Western Blot (WB)||See 1 publications below|
PA1-338 detects SREBP 2 in human, mouse and rat samples.
PA1-338 has been successfully used in Western blot and ICC/IF procedures. By Western blot, this antibody detects an ~68 and 120 kDa protein representing SREBP 2 in mouse and rat liver samples as well as rat kidney samples. A predominent band at ~68 kDa (active cleaved site) is seen and a band at ~120 kDa (inactive precursor) may not be seen or it may be diminished.
The PA1-338 immunogen is a synthetic peptide corresponding to residues A(843) I S W L Q G D D A A V R S H(857) of rat SREBP 2. This sequence is identical in human and differs by 1 a.a. in mouse. PA1-338 immunizing peptide (Cat. # PEP-275) is available for use in neutralization and control experiments.
Sterol regulatory element-binding proteins (SREBPs) are transcription factors that are members of the basic helix-loop-helix leucine zipper family of DNA binding proteins. Three isoforms have been identified in mammalian tissues that vary in structure, regulation, and function. SREBP-1a and SREBP-1c (originally cloned as ADD1) are protein products of alternative promoter usage of the SREBP-1 gene. The third isoform is transcribed from a different gene, SREBP-2.
SREBPs are present as 120 kDa inactive precursors in the endoplasmic reticulum (ER) membrane. Upon activation, the SREBP protein is translocated to the Golgi and proteolytic cleavage occurs resulting in a mature transcriptionally active 60-78 kDa fragment.
In liver and adipose tissues, SREBPs have a significant influence on lipid and cholesterol accumulation by inducing the transcription of genes involved in these processes. While SREBP-1 is thought to be more important in regulating the expression of genes involved in triglyceride synthesis and accumulation, SREBP-2 has been more closely linked to those involved in cholesterol synthesis and accumulation.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
High copper selectively alters lipid metabolism and cell cycle machinery in the mouse model of Wilson disease.
PA1-338 was used in western blot to investigate the role of SREBP2 during the copper-induced alteration of lipid metabolism and cell cycle machinery.
|Huster D,Purnat TD,Burkhead JL,Ralle M,Fiehn O,Stuckert F,Olson NE,Teupser D,Lutsenko S||The Journal of biological chemistry (282:8343)||2007|