WB result of Histone H3 (Lactyl K9) Recombinant Rabbit mAb
Primary antibody: Histone H3 (Lactyl K9) Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: untreated HeLa whole cell lysate 20 µg
Lane 2: HeLa treated with 100 mM Lactate sodium for 24 hours whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 15 kDa
Observed MW: 17 kDa
Histone H3 (Lactyl K9) Recombinant Rabbit mAb (S-R397)
Histone H3 (Lactyl K9) Recombinant Rabbit mAb (S-R397)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | Histone H3 (Lactyl K9) |
| Synonyms | H3K9lac,H3K9la |
| Location | Nucleus |
| Accession | P68431 |
| Clone Number | S-R397 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, ICC, IP, ChIP |
| Reactivity | Hu |
| Purification | Protein A |
| Concentration | 0.5 mg/ml |
| Conjugation | Unconjugated |
| Physical Appearance | Liquid |
| Storage Buffer | PBS, 40% Glycerol, 0.05% BSA, 0.03% Proclin 300 |
| Stability & Storage | 12 months from date of receipt / reconstitution, -20 °C as supplied |
Dilution
| application | dilution | species |
| WB | 1:1000 | |
| IP | 1:50 | |
| ICC | 1:500 | |
| ChIP | 1:20-1:50 |
Background
H3K9lac refers to the lactylation modification that occurs at the 9th lysine residue of histone H3. This modification is a relatively novel histone modification discovered in recent epigenetic studies. This modification may alter the interaction between histones, DNA, and other proteins, thereby affecting gene expression and cellular functions.
Picture
Picture
Western Blot
IP
Histone H3 (Lactyl K9) Rabbit mAb at 1/50 dilution (1 µg) immunoprecipitating Histone H3 (Lactyl K9) in 0.4 mg HeLa treated with 100 mM Lactate sodium for 24 hours whole cell lysate.
Western blot was performed on the immunoprecipitate using Histone H3 (Lactyl K9) Rabbit mAb at 1/1000 dilution.
Secondary antibody (HRP) for IP was used at 1/1000 dilution.
Lane 1: HeLa treated with 100 mM Lactate sodium for 24 hours whole cell lysate 20 µg (Input)
Lane 2: Histone H3 (Lactyl K9) Rabbit mAb IP in HeLa treated with 100 mM Lactate sodium for 24 hours whole cell lysate
Lane 3: Rabbit monoclonal IgG IP in HeLa treated with 100 mM Lactate sodium for 24 hours whole cell lysate
Predicted MW: 15 kDa
Observed MW: 17 kDa
Immunocytochemistry
ICC analysis of HeLa cells treated with sodium lactate (100 mM, 24 hours) (top panel) and HeLa cells untreated with sodium lactate (100 mM, 24 hours) (below panel). Anti- Histone H3 (Lactyl K9) antibody was used at 1/500 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 100% ice-cold methanol and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (Red).
ChIP
Chromatin immunoprecipitation (ChIP) was performed on HeLa cells + Lactate sodium (100 mM, 24 hours) cells cross - linked with 1% formaldehyde for 10 min, then chromatin was fragmented by sonication. Parallel reactions used Histone H3 (Lactyl K9) Recombinant Rabbit mAb (S-R397) and Rabbit mAb IgG Isotype Control (SDT-R173) at 1:50 for immunoprecipitation.
Post - immunoprecipitation, both samples were washed, eluted, and cross - links reversed. Purified DNA was analyzed by qPCR.
qPCR (%input: immunoprecipitated DNA/input DNA)
showed the enrichment of RPL30, GAPDH, MYOD1,
AFM, SAT-α and SAT-2 in Histone H3 (Lactyl K9) Recombinant
Rabbit mAb (S-R397)-immunoprecipitated sample.
