WB
Cell treatment (CT),Relative expression (RE) Western blot analysis of Histone H3.3 (mutated G34V) on different lysates with Rabbit anti-Histone H3.3 (mutated G34V) antibody at 1/2,000 dilution. Lysates/proteins at 20 µg/Lane. Exposure time: 59 seconds; Blocking: 5% NFDM/TBST, 1 hour at room temperature. Primary antibody: Rabbit anti-Histone H3.3 (mutated G34V) antibody, 1/2,000 in primary antibody dilution buffer, overnight at 4 °C. Secondary antibody: Goat anti-Rabbit IgG-HRP, 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature.ICC/IF
Cell treatment (CT),Relative expression (RE) Application: Immunocytochemistry (IF-cell) Species: Human Sample: Wild-type 293T 293T (Human embryonic kidney cell) transfected with Histone H3.3 (mutated G34R) 293T (Human embryonic kidney cell) transfected with Histone H3.3 (mutated G34V) Fixation: 4% Paraformaldehyde, 15 minutes at room temperature. Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature. Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature. Antibody dilution buffer: 1% BSA in PBST. Primary antibody: 1/5,000, overnight at 4℃. Secondary antibody: Goat Anti-Rabbit IgG (488), 45 minutes at room temperature. Counterstain: Beta tubulin (Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).IHC
Cell treatment (CT) Application: Immunohistochemistry (IHC-P) Species: Human Tissue: 293T transfected with Histone H3.3 G34V expression vector Sample: Paraffin-embedded section Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C. Wash buffer: 1× TBST Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature. Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature. Primary antibody: 1/1,000, 1 hour at room temperature. Secondary antibody: 20 minutes at room temperature.IP
Cell treatment (CT) Immunoprecipitation (IP) Histone H3.3 (mutated G34V) was immunoprecipitated in 0.2 mg 293T (Human embryonic kidney cell) transfected with Histone H3.3 G34V expression vector containing a Flag-tag cell lysate cell lysate with Histone H3.3 (mutated G34V) Recombinant Rabbit Monoclonal Antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using Histone H3.3 (mutated G34V) Recombinant Rabbit Monoclonal Antibody at 1/5,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/1,000 dilution was used for 1 hour at room temperature. Exposure time: 14 seconds. Blocking: 5% NFDM/TBST, 1 hour at room temperature. Primary dilution: 1/1,000 in primary antibody dilution buffer, 2 hours at room temperature.| Product Name | Histone H3.3 (mutated G34V) Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | ICC/IFIHCIPWB |
| WB:1:2000 ICC/IF:1:5000 IHC:1:1000 IP:1-2μg/sample |
|
| Species Reactivity | Human |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | H3-3A |
|---|---|
| Gene Synonyms | H3F3 H3.3A H3F3A BRYLIB1 |
| Gene Full Name | H3.3 histone A |
| Gene Summary | Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Two molecules of each of the four core histones (H2A, H2B, H3, and H4) form an octamer, around which approximately 146 bp of DNA is wrapped in repeating units, called nucleosomes. The linker histone, H1, interacts with linker DNA between nucleosomes and functions in the compaction of chromatin into higher order structures. This gene contains introns and its mRNA is polyadenylated, unlike most histone genes. The protein encoded is a replication-independent member of the histone H3 family. [provided by RefSeq, Jul 2008] |
| Molecular Weight(MW) | 15 kDa(Observed band size: 20 kDa) |
| Cellular Localization | Nucleus, Chromosome. |
| SwissProt ID | P84243 |
|---|
WB
Cell treatment (CT),Relative expression (RE) Western blot analysis of Histone H3.3 (mutated G34V) on different lysates with Rabbit anti-Histone H3.3 (mutated G34V) antibody at 1/2,000 dilution. Lysates/proteins at 20 µg/Lane. Exposure time: 59 seconds; Blocking: 5% NFDM/TBST, 1 hour at room temperature. Primary antibody: Rabbit anti-Histone H3.3 (mutated G34V) antibody, 1/2,000 in primary antibody dilution buffer, overnight at 4 °C. Secondary antibody: Goat anti-Rabbit IgG-HRP, 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature.
ICC/IF
Cell treatment (CT),Relative expression (RE) Application: Immunocytochemistry (IF-cell) Species: Human Sample: Wild-type 293T 293T (Human embryonic kidney cell) transfected with Histone H3.3 (mutated G34R) 293T (Human embryonic kidney cell) transfected with Histone H3.3 (mutated G34V) Fixation: 4% Paraformaldehyde, 15 minutes at room temperature. Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature. Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature. Antibody dilution buffer: 1% BSA in PBST. Primary antibody: 1/5,000, overnight at 4℃. Secondary antibody: Goat Anti-Rabbit IgG (488), 45 minutes at room temperature. Counterstain: Beta tubulin (Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).
IHC
Cell treatment (CT) Application: Immunohistochemistry (IHC-P) Species: Human Tissue: 293T transfected with Histone H3.3 G34V expression vector Sample: Paraffin-embedded section Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C. Wash buffer: 1× TBST Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature. Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature. Primary antibody: 1/1,000, 1 hour at room temperature. Secondary antibody: 20 minutes at room temperature.
IP
Cell treatment (CT) Immunoprecipitation (IP) Histone H3.3 (mutated G34V) was immunoprecipitated in 0.2 mg 293T (Human embryonic kidney cell) transfected with Histone H3.3 G34V expression vector containing a Flag-tag cell lysate cell lysate with Histone H3.3 (mutated G34V) Recombinant Rabbit Monoclonal Antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using Histone H3.3 (mutated G34V) Recombinant Rabbit Monoclonal Antibody at 1/5,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/1,000 dilution was used for 1 hour at room temperature. Exposure time: 14 seconds. Blocking: 5% NFDM/TBST, 1 hour at room temperature. Primary dilution: 1/1,000 in primary antibody dilution buffer, 2 hours at room temperature.| Application Notes | WB:1:2000 ICC/IF:1:5000 IHC:1:1000 IP:1-2μg/sample |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C long term. Avoid freeze / thaw cycle. |
| Storage Buffer | 1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950. |
