Anti-RNA polymerase II Antibody, clone CTD4H8 ZooMAb® Mouse Monoclonal
SIGMA/ZMS1031 - recombinant, expressed in HEK 293 cells
Synonym: DNA-directed RNA polymerase II subunit A; DNA-directed RNA polymerase II subunit RPB1; DNA-directed RNA polymerase III largest subunit; EC:2.7.7.6; RNA polymerase II subunit B1; RNA-directed RNA polymerase II subunit RPB1
Product Type: Chemical
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human cerebral cortex (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZMS1031, Anti-RNA polymerase II, clone CTD4H8 ZooMAb® Mouse Monoclonal. Reactivity was detected using an Rabbit Anti-Mouse IgG and HRP-DAB. Nuclear staining was observed in subsets of neurons, glial cells and endothelial cells of human cerebral cortex tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human cerebral cortex (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZMS1031, Anti-RNA polymerase II, clone CTD4H8 ZooMAb® Mouse Monoclonal. Reactivity was detected using an Rabbit Anti-Mouse IgG and HRP-DAB. Nuclear staining was observed in subsets of neurons, glial cells and endothelial cells of human cerebral cortex tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human cerebral cortex (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZMS1031, Anti-RNA polymerase II, clone CTD4H8 ZooMAb® Mouse Monoclonal. Reactivity was detected using an Rabbit Anti-Mouse IgG and HRP-DAB. Nuclear staining was observed in subsets of neurons, glial cells and endothelial cells of human cerebral cortex tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human testis (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZMS1031, Anti-RNA polymerase II, clone CTD4H8 ZooMAb® Mouse Monoclonal. Reactivity was detected using an Rabbit Anti-Mouse IgG and HRP-DAB. Nuclear staining was also observed in seminiferous tubules and Leydig cells of human testis tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human testis (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZMS1031, Anti-RNA polymerase II, clone CTD4H8 ZooMAb® Mouse Monoclonal. Reactivity was detected using an Rabbit Anti-Mouse IgG and HRP-DAB. Nuclear staining was also observed in seminiferous tubules and Leydig cells of human testis tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human testis (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZMS1031, Anti-RNA polymerase II, clone CTD4H8 ZooMAb® Mouse Monoclonal. Reactivity was detected using an Rabbit Anti-Mouse IgG and HRP-DAB. Nuclear staining was also observed in seminiferous tubules and Leydig cells of human testis tissue sections.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of HeLa cells was performed using a 1:100 dilution of Cat. No. ZMS1031, Anti-RNA polymerase II, clone CTD4H8 ZooMAb® Mouse Monoclonal and visualized with a Goat Anti-Mouse secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the nucleus.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of HeLa cells was performed using a 1:100 dilution of Cat. No. ZMS1031, Anti-RNA polymerase II, clone CTD4H8 ZooMAb® Mouse Monoclonal and visualized with a Goat Anti-Mouse secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the nucleus.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of HeLa cells was performed using a 1:100 dilution of Cat. No. ZMS1031, Anti-RNA polymerase II, clone CTD4H8 ZooMAb® Mouse Monoclonal and visualized with a Goat Anti-Mouse secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the nucleus.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of A431 cells was performed using a 1:100 dilution of Cat. No. ZMS1031, Anti-RNA polymerase II, clone CTD4H8 ZooMAb® Mouse Monoclonal and visualized with a Goat Anti-Mouse secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the nucleus.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of A431 cells was performed using a 1:100 dilution of Cat. No. ZMS1031, Anti-RNA polymerase II, clone CTD4H8 ZooMAb® Mouse Monoclonal and visualized with a Goat Anti-Mouse secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the nucleus.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of A431 cells was performed using a 1:100 dilution of Cat. No. ZMS1031, Anti-RNA polymerase II, clone CTD4H8 ZooMAb® Mouse Monoclonal and visualized with a Goat Anti-Mouse secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the nucleus.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of HUVEC was performed using a 1:100 dilution of Cat. No. ZMS1031, Anti-RNA polymerase II, clone CTD4H8 ZooMAb® Mouse Monoclonal and visualized with a Goat Anti-Mouse secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the nucleus.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of HUVEC was performed using a 1:100 dilution of Cat. No. ZMS1031, Anti-RNA polymerase II, clone CTD4H8 ZooMAb® Mouse Monoclonal and visualized with a Goat Anti-Mouse secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the nucleus.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of HUVEC was performed using a 1:100 dilution of Cat. No. ZMS1031, Anti-RNA polymerase II, clone CTD4H8 ZooMAb® Mouse Monoclonal and visualized with a Goat Anti-Mouse secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the nucleus.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of NIH3T3 cells was performed using a 1:100 dilution of Cat. No. ZMS1031, Anti-RNA polymerase II, clone CTD4H8 ZooMAb® Mouse Monoclonal and visualized with a Goat Anti-Mouse secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the nucleus.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of NIH3T3 cells was performed using a 1:100 dilution of Cat. No. ZMS1031, Anti-RNA polymerase II, clone CTD4H8 ZooMAb® Mouse Monoclonal and visualized with a Goat Anti-Mouse secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the nucleus.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of NIH3T3 cells was performed using a 1:100 dilution of Cat. No. ZMS1031, Anti-RNA polymerase II, clone CTD4H8 ZooMAb® Mouse Monoclonal and visualized with a Goat Anti-Mouse secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the nucleus.
Western Blotting Enhanced Validation-Recombinant Antibody Technology Lysates from HeLa cells (Lane 1) and L6 cells (Lane 2) were probed with Cat. No. ZMS1031, Anti-RNA polymerase II, clone CTD4H8 ZooMAb® Mouse Monoclonal (1:1,000 dilution). Proteins were visualized using a Goat Anti-Mouse IgG1 secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates RNA polymerase II (~300 kDa).
Western Blotting Enhanced Validation-Recombinant Antibody Technology NIH3T3 cell lysate was probed with Cat. No. ZMS1031, Anti-RNA polymerase II, clone CTD4H8 ZooMAb® Mouse Monoclonal (1:1,000 dilution). Proteins were visualized using a Goat Anti-Mouse IgG1 secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates RNA polymerase II (~300 kDa).
Flow Cytometry Enhanced Validation-Recombinant Antibody Technology Staining of one million HUVEC was performed using 0.1 μg of Cat. No. ZMS1031, Anti-RNA polymerase II, clone CTD4H8 ZooMAb® Mouse Monoclonal (Yellow histogram) or the equivalent amount of a Mouse IgG1 isotype control (Grey histogram), followed by a PE-conjugated Goat Anti-MouseIgG1 secondary antibody.
12 Principles of Green Chemistry: Principle 1—Waste Prevention. This product has a smaller environmental footprint through reduction of waste in the manufacturing process.
12 Principles of Green Chemistry: Principle 4—Designing Safer Chemicals
Chemical products should be designed to affect their desired function while minimizing their toxicity.
Chemical products should be designed to affect their desired function while minimizing their toxicity.
12 Principles of Green Chemistry: Principle 6—Design for Energy Efficiency. This product was re-engineered to utilize less energy in the manufacturing process.
| antibody form | purified antibody |
| antibody product type | primary antibodies |
| biological source | mouse |
| clone | CTD4H8, monoclonal |
| recombinant monoclonal | |
| conjugate | unconjugated |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation  |
| form | lyophilized |
| greener alternative category | Aligned |
| greener alternative product characteristics | Waste Prevention Designing Safer Chemicals Design for Energy Efficiency Learn more about the Principles of Green Chemistry . |
| isotype | IgG1 |
| mol wt | calculated mol wt 217.18 kDa |
| observed mol wt ~300 kDa | |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| Quality Level | 200 |
| recombinant | expressed in HEK 293 cells |
| shipped in | ambient |
| species reactivity | human |
| species reactivity (predicted by homology) | rat, mouse, yeast |
| storage temp. | 2-8°C |
| technique(s) | flow cytometry: suitable |
| immunocytochemistry: suitable | |
| immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable | |
| western blot: suitable | |
| UniProt accession no. | P24928  |
| Application: | Anti-RNA polymerase II, clone CTD4H8 ZooMAb, Cat. No. ZMS1031, is a recombinant Mouse monoclonal antibody that detects RNA polymerase II and is tested in Flow Cytometry, Immunocytochemistry, Immunohistochemistry (Paraffin), and Western Blotting. |
| Application: | Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected RNA polymerase II in HeLa and NIH3T3 cell lysate. Flow Cytometry Analysis: 0.1 μg from a representative lot detected RNA polymerase II in one million HUVEC. Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected RNA polymerase II in human cerebral cortex and human testis tissue sections. Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected RNA polymerase II in HeLa, A431, HUVEC, and NIH 3T3 cells. Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user |
| Disclaimer: | Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals. |
| General description: | We are committed to bringing you greener alternative products, which adhere to one or more of The 12 Principles of Green Chemistry.This antibody is Preservative-free, produced without the harm or sacrifice of animals and exceptionally stable to allow for ambient shipping and storage if needed and thus aligns with "Waste Prevention", "Designing Safer Chemicals" and "Design for Energy Efficiency". Click here for more information. |
| General description: | ZooMAb antibodies represent an entirely new generation of recombinant monoclonal antibodies. Each ZooMAb antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb antibodies are reliably available and ready to ship when you need them. Learn more about ZooMAb here. |
| Immunogen: | Peptide containing 10 repeats of YSPT[pS]PS corresponding to the C-terminal region of human RNA polymerase II. |
| Legal Information: | ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany |
| Physical form: | Purified recombinant mouse monoclonal antibody IgG, lyophilized in PBS, 5% Trehalose, normal appearance a coarse or translucent resin. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 μL. |
| Reconstitution: | 30 μg/mL after reconstitution at 25 μL per vial. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type. |
| Specificity: | Clone CTD4H8 is a ZooMAb mouse recombinant monoclonal antibody that specifically detects RNA polymerase II. It targets an epitope within the C-terminal region. |
| Storage and Stability: | Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 μL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws. |
| Target description: | DNA-directed RNA polymerase II subunit RPB1 (UniProt: P24928; also known as EC:2.7.7.6; RNA polymerase II subunit B1, DNA-directed RNA polymerase II subunit A, DNA-directed RNA polymerase III largest subunit, RNA-directed RNA polymerase II subunit RPB1, POLR2A) is encoded by the POLR2A (also known as POLR2) gene (Gene ID: 5430) in human. DNA-directed RNA polymerase II subunit RPB1 is the catalytic component of RNA polymerase II that catalyzes the transcription of DNA into RNA using the four ribonucleoside triphosphates as substrates. It has multiple Zinc and magnesium ion binding sites. The binding of ribonucleoside triphosphate to the RNA polymerase II transcribing complex involves a two-step mechanism. The initial binding seems to occur at the entry (E) site and involves a magnesium ion temporarily coordinated by three conserved aspartate residues of the two largest RNA Pol II subunits. The ribonucleoside triphosphate is transferred by a rotation to the nucleotide addition (A) site for pairing with the template DNA. The catalytic A site involves three conserved aspartate residues of the RNA Pol II largest subunit which permanently coordinate a second magnesium ion. POLR2A is localized to cytoplasm in its hypo-phosphorylated form and the active hyper-phosphorylated form is located mainly in the nucleus. The tandem heptapeptide repeats (Tyr1-Ser2-Pro3-Thr4-Ser5 |
| WGK Germany | WGK 1 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | 2-8°C |
| UNSPSC | 12352203 |