WB
Blocking buffer: 5% NFDM/TBST Primary ab dilution: 1:2000 Primary ab incubation condition: 2 hours at room temperature Secondary ab: Goat Anti-Rabbit IgG H&L (HRP) Lysate: HeLa, NIH/3T3, BRL Protein loading quantity: 20 μg Exposure time: 60 s.ChIP
Cell type: HeLa+Sodium lactate (100 mM, 24hr) Cross-linking conditions: No cross-linking Amount of chromatin per IP: 5×106 cells Amount of Ab per IP: 6 ug Beads type and amount per IP: 50 μl of Protein A/G MagBeads Comment: The ChIP was performed with 6 μg of normal rabbit IgG as a negative control. Real time quantitative PCR was performed on immunoprecipitated DNA using primers specific for the human GAPDH CDS region, RPL30, LDHA-promoter, LDHA CDS region, FOXO3apromoter, FOXO3a-downstream, RAB20 and TuBBP10. Data are presented as enrichment of each sample relative to total amount of input chromatin at each amplicon.Cut&tag
Cell type: HeLa + sodium lactate (25 mM, 24hr) Quantity of cell: 1×105 cells Primary ab dilution: 1: 100 Primary ab incubation condition: 4℃ overnight Secondary ab: Goat Anti-Rabbit IgG H&L pAb Secondary ab dilution: 1: 1400.| Product Name | Anti-L-Lactyl-Histone H4 (Lys8) Rabbit mAb |
|---|---|
| Antibody Type | Primary Antibodies |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | ChIPWBCUT&Tag-seq |
| WB:1:500-1:2000 CHIP:6 ug/5x106 cells Cut&tag:1:100 |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | H4K8la |
|---|---|
| Molecular Weight(MW) | 15kDa |
| Function | Histones are subject to a variety of enzyme catalyzed modifications, including acetylation, methylation, phosphorylation, ubiquitylation, etc. The Warburg effect, which originally described increased production of lactate in cancer, is associated with diverse cellular processes such as angiogenesis, hypoxia, polarization of macrophages and activation of T cells. This phenomenon is intimately linked to several diseases including neoplasia, sepsis and autoimmune diseases. Lactate, which is converted from pyruvate in tumour cells, is widely known as an energy source and metabolic by-product. Zhang et al. (Nature, 2019) reveal that histone lactylation is a previously unknown histone modification, derived from the cellular metabolite lactate. |
| Cellular Localization | Nucleus. |
WB
Blocking buffer: 5% NFDM/TBST Primary ab dilution: 1:2000 Primary ab incubation condition: 2 hours at room temperature Secondary ab: Goat Anti-Rabbit IgG H&L (HRP) Lysate: HeLa, NIH/3T3, BRL Protein loading quantity: 20 μg Exposure time: 60 s.
ChIP
Cell type: HeLa+Sodium lactate (100 mM, 24hr) Cross-linking conditions: No cross-linking Amount of chromatin per IP: 5×106 cells Amount of Ab per IP: 6 ug Beads type and amount per IP: 50 μl of Protein A/G MagBeads Comment: The ChIP was performed with 6 μg of normal rabbit IgG as a negative control. Real time quantitative PCR was performed on immunoprecipitated DNA using primers specific for the human GAPDH CDS region, RPL30, LDHA-promoter, LDHA CDS region, FOXO3apromoter, FOXO3a-downstream, RAB20 and TuBBP10. Data are presented as enrichment of each sample relative to total amount of input chromatin at each amplicon.
Cut&tag
Cell type: HeLa + sodium lactate (25 mM, 24hr) Quantity of cell: 1×105 cells Primary ab dilution: 1: 100 Primary ab incubation condition: 4℃ overnight Secondary ab: Goat Anti-Rabbit IgG H&L pAb Secondary ab dilution: 1: 1400.| Application Notes | WB:1:500-1:2000 CHIP:6 ug/5x106 cells Cut&tag:1:100 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at -20°C. Stable for one year after shipment. |
| Storage Buffer | PBS with 0.02% sodium azide and 50% glycerol pH 7.3. |
