WB result of RNA Polymerase II Recombinant Rabbit mAb
Primary antibody: RNA Polymerase II Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: 293T whole cell lysate 20 µg
Lane 2: HeLa whole cell lysate 20 µg
Lane 3: MCF7 whole cell lysate 20 µg
Lane 4: HCT 116 whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 217 kDa
Observed MW: 180~250 kDa
RNA Polymerase II Recombinant Rabbit mAb (S-969-102)
RNA Polymerase II Recombinant Rabbit mAb (S-969-102)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | RNA Polymerase II |
| Synonyms | DNA-directed RNA polymerase II subunit RPB1; RNA polymerase II subunit B1; 3'-5' exoribonuclease; DNA-directed RNA polymerase II subunit A; DNA-directed RNA polymerase III largest subunit; RNA-directed RNA polymerase II subunit RPB1; POLR2A; POLR2 |
| Immunogen | Synthetic Peptide |
| Location | Cytoplasm, Nucleus |
| Accession | P24928 |
| Clone Number | S-969-102 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, ICC, IP, ChIP |
| Reactivity | Hu, Ms, Rt |
| Positive Sample | 293T, HeLa, MCF7, HCT 116, NIH/3T3, RAW264.7, C2C12, C6 |
| Predicted Reactivity | Ba |
| 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 | Hu, Ms, Rt |
| IP | 1:50 | Hu |
| ICC | 1:500 | Hu |
| ChIP | 1:20-1:50 | Hu |
Background
RNA Polymerase II (RNAP II) is a crucial enzyme in eukaryotic cells responsible for transcribing protein-coding genes into mRNA and some non-coding RNAs. The function of RNAP II is to catalyze the transcription of DNA into RNA, which is a critical step in gene expression. The process of transcription can be divided into three stages: initiation, elongation, and termination. During initiation, RNAP II binds to the promoter region of a gene, assisted by various transcription factors, and begins to synthesize RNA. Elongation follows, where RNAP II moves along the DNA template, synthesizing the RNA chain. Termination occurs when RNAP II reaches a stop signal, releasing the newly synthesized RNA and disengaging from the DNA template. RNAP II plays a significant role in the regulation of transcription. It works in conjunction with a set of general transcription factors (GTFs), including TFIIA, TFIIB, TFIID, TFIIE, TFIIF, and TFIIH, to coordinate the transcription process. These factors dynamically associate with and dissociate from RNAP II to regulate the occurrence of transcription. RNAP II is also involved in the processing of mRNA, such as capping, splicing, and polyadenylation, through interactions with various factors during these modifications. Additionally, the CTD of RNAP II can undergo a series of modifications, such as phosphorylation, which provides a rapid pathway for plants to activate primary immune responses in the event of pathogen invasion.
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Western Blot
WB result of RNA Polymerase II Recombinant Rabbit mAb
Primary antibody: RNA Polymerase II Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: NIH/3T3 whole cell lysate 20 µg
Lane 2: RAW264.7 whole cell lysate 20 µg
Lane 3: C2C12 whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 217 kDa
Observed MW: 180~250 kDa
WB result of RNA Polymerase II Recombinant Rabbit mAb
Primary antibody: RNA Polymerase II Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: C6 whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 217 kDa
Observed MW: 180~250 kDa
IP
RNA Polymerase II Recombinant Rabbit mAb at 1/50 dilution (1 µg) immunoprecipitating RNA Polymerase II in 0.4 mg 293T whole cell lysate.
Western blot was performed on the immunoprecipitate using RNA Polymerase II Recombinant Rabbit mAb at 1/1000 dilution.
Secondary antibody (HRP) for IP was used at 1/1000 dilution.
Lane 1: 293T whole cell lysate 40 µg (Input)
Lane 2: RNA Polymerase II Recombinant Rabbit mAb IP in 293T whole cell lysate
Lane 3: Rabbit monoclonal IgG IP in 293T whole cell lysate
Predicted MW: 217 kDa
Observed MW: 180~250 kDa
Exposure time: 180 s
Immunocytochemistry
ICC shows positive staining in HeLa cells. Anti- RNA Polymerase II 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 4% PFA 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 cross - linked with 1% formaldehyde for 10 min, then chromatin was fragmented by sonication. Parallel reactions used RNA Polymerase II Recombinant Rabbit mAb (S-969-102) 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 GAPDH, AFM and SAT-α in RNA Polymerase II Recombinant Rabbit mAb (S-969-102) - immunoprecipitated sample.
