Lactyl-Histone H3 (K18) Recombinant Rabbit Monoclonal Antibody [PSH26-77]
cat.: HA752044
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse |
| Applications: | WB, IP, Dot Blot, ChIP |
| Clonality: | Monoclonal |
| Clone number: | PSH26-77 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | PBS (pH7.4). |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 15 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic Lactyl-peptide corresponding to residues surrounding Lys18 of human Histone H3. |
| Positive control: | HeLa (Human cervical adenocarcinoma cell) cell lysate, HeLa treated with 100mM Lactate sodium for 24 hours cell lysate, HT-29 (Human colorectal adenocarcinoma cell) cell lysate, HT-29 treated with 100mM Lactate sodium for 24 hours cell lysate, NIH/3T3 (Mouse fibroblast) cell lysate, NIH/3T3 treated with 100mM Lactate sodium for 24 hours cell lysate. |
| Subcellular location: | Nucleus. |
| Recommended Dilutions:
WB IP Dot Blot ChIP |
1:5,000 1-2μg/sample 1:5,000 Use 5 μg for 25 μg of chromatin. |
| Uniprot #: | SwissProt: P68431 Human | P68433 Mouse |
| Alternative names: | H3 histone family member E pseudogene H3 histone family, member A H3/A H31_HUMAN H3F3 H3FA Hist1h3a HIST1H3B HIST1H3C HIST1H3D HIST1H3E HIST1H3F HIST1H3G HIST1H3H HIST1H3I HIST1H3J HIST3H3 histone 1, H3a Histone cluster 1, H3a Histone H3 3 pseudogene Histone H3.1 Histone H3/a Histone H3/b Histone H3/c Histone H3/d Histone H3/f Histone H3/h Histone H3/i Histone H3/j Histone H3/k Histone H3/l |
Images
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Fig1:
Western blot analysis of Lactyl-Histone H3 (K18) on different lysates with Rabbit anti-Lactyl-Histone H3 (K18) antibody (HA752044) at 1/5,000 dilution.
Lane 1: HeLa (Human cervical adenocarcinoma cell) cell lysate Lane 2: HeLa treated with 100mM Lactate sodium for 24 hours cell lysate Lane 3: HT-29 (Human colorectal adenocarcinoma cell) cell lysate Lane 4: HT-29 treated with 100mM Lactate sodium for 24 hours cell lysate Lysates/proteins at 20 µg/Lane. Exposure time: 3 minutes; ECL: K1801 Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary antibody: HA752044, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 °C Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature Predicted band size: 15 kDa Observed band size: 15 kDa |
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Fig2:
Western blot analysis of Lactyl-Histone H3 (K18) on different lysates with Rabbit anti-Lactyl-Histone H3 (K18) antibody (HA752044) at 1/5,000 dilution.
Lane 1: NIH/3T3 (Mouse fibroblast) cell lysate Lane 2: NIH/3T3 treated with 100mM Lactate sodium for 24 hours cell lysate Lysates/proteins at 15 µg/Lane. Exposure time: 42 seconds; ECL: K1801 Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary antibody: HA752044, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 °C Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature Predicted band size: 15 kDa Observed band size: 15 kDa |
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Fig3:
Immunoprecipitation (IP)
Lactyl-Histone H3 (K18) was immunoprecipitated in 0.2 mg HeLa (Human cervix adenocarcinoma epithelial cell) treated with 100mM Lactate sodium for 24 hours cell lysate with HA752044 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA752044 at 1/5,000 dilution. Anti-Rabbit IgG for IP, AlpSdAbs® VHH(HRP) (025-100-005) at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: HeLa treated with 100mM Lactate sodium for 24 hours cell lysate (input) Lane 2: HA752044 IP in HeLa treated with 100mM Lactate sodium for 24 hours cell lysate Lane 3: Rabbit IgG instead of HA752044 in HeLa treated with 100mM Lactate sodium for 24 hours cell lysate Exposure time: 3 minutes Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary dilution: HA752044, 1/5,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature Predicted band size: 15 kDa Observed band size: 15 kDa |
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Fig4:
Dot blot analysis of Lactyl-Histone H3 (K18) on different peptides with Rabbit anti-Lactyl-Histone H3 (K18) antibody (HA752044) at 1/5,000 dilution.
Lane 1: Lactyl-Histone H3 (Lys18) Lane 2: Acetyl-Histone H3 (Lys18) Lane 3: Unmodified Histone H3 Proteins loading: 100ng, 50ng, 10ng, 5ng; Exposure time: 20 seconds; ECL: K1801 Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary antibody: HA752044, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 °C Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature |
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Fig5: Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells treated with 100nM Lactate sodium for 24 hours with Lactyl-Histone H3 (K18) (HA752044) / Competitor's antibody / Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one. |
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