HMGB1 (High Mobility Group Box-1)

Size

£876.00

SKU:: 353417
Purity:: Purified by Protein G affinity chromatography.
Storage Conditions:: -20°C
Supplier:: United States Biological
Host:: Hamster
Immunogen:: Recombinant protein corresponding to HMGB1.
Clone:: 5H6
Format:: Supplied as a liquid in PBS, 1% BSA, 0.02% sodium azide.
Extra Details:: HMGB1 (High Mobility Group Box-1 protein also known as amphoterin) is a highly conserved protein with more than 95% aa identity between rodent and human HMGB1. It is a prevalent non-histone chromatin component and a non-sequence specific DNA binding protein. HMGB1 consists of two homologous HMG boxes rich in basic amino acids and an acidic tail at the carboxy-terminus. HMGB1 is involved in the regulation of chromatin structure as well as being involved, either as positive or negative factors with various aspects of DNA replication, transcription, repair, and ligation. HMGB1, identified as a membrane associated protein termed “amphoterin,” mediates neurite outgrowth, tumor outgrowth, and metastasis. It participates in plasminogen activation and is recognized as a late mediator of endotoxin lethality in mice. Applications: Suitable for use in Immunofluorescence, Flow Cytometry and Western Blot. Other applications not tested. Recommended Dilution: Immunofluorescence: NIH-3T3 cells were fixed with 2% PFA (15 min). After fixation, cells were washed with PBS and incubated for 5 min at 4ºC with permeabilization buffer. The cells were blocked with 1% BSA/10% goat serum in PBS for 10 min and were incubated with HMGB1 (10ug/ml) overnight at 4ºC. The secondary antibody was biotinylated goat anti-hamster IgG at a 1:500 dilution for 1 hour. Finally, streptavidin was used at a 1:500 dilution for 30 min. Flow Cytometry: For staining cells were permeabilized with buffer containing 0.5% saponin. The cells were fixed in 0.01% formaldehyde before staining. Hamster IgG isotype control is used as a negative control. Western Blot: A band is expected at ~30kD using reducing conditions. For non-reducing conditions a band is expected at both 27kD and 30kD. Optimal dilutions to be determined by the researcher. Storage and Stability: May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

Shipping Conditions:: Blue Ice