Anti-POLR2A Antibody, clone 3G21 ZooMAb® Rabbit Monoclonal
SIGMA/ZRB1409 - 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 testis (A) and Negative control (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1409, Anti-POLR2A, clone 3G21 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit and HRP-DAB. Nuclear staining was observed in seminiferous tubules of human testis tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human testis (A) and Negative control (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1409, Anti-POLR2A, clone 3G21 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit and HRP-DAB. Nuclear staining was observed in seminiferous tubules of human testis tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human testis (A) and Negative control (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1409, Anti-POLR2A, clone 3G21 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit and HRP-DAB. Nuclear staining was observed in seminiferous tubules of human testis tissue sections.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HeLa cell lines were performed using a 1:100 dilution of Cat. No. ZRB1409, Anti-POLR2A, clone 3G21 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit 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 cell lines were performed using a 1:100 dilution of Cat. No. ZRB1409, Anti-POLR2A, clone 3G21 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit 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 cell lines were performed using a 1:100 dilution of Cat. No. ZRB1409, Anti-POLR2A, clone 3G21 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit 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 cell lines were performed using a 1:100 dilution of Cat. No. ZRB1409, Anti-POLR2A, clone 3G21 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit 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 cell lines were performed using a 1:100 dilution of Cat. No. ZRB1409, Anti-POLR2A, clone 3G21 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit 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 cell lines were performed using a 1:100 dilution of Cat. No. ZRB1409, Anti-POLR2A, clone 3G21 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit 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 cell lines were performed using a 1:100 dilution of Cat. No. ZRB1409, Anti-POLR2A, clone 3G21 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit 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 cell lines were performed using a 1:100 dilution of Cat. No. ZRB1409, Anti-POLR2A, clone 3G21 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit 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 cell lines were performed using a 1:100 dilution of Cat. No. ZRB1409, Anti-POLR2A, clone 3G21 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit 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 NIH/3T3 cell lines were performed using a 1:100 dilution of Cat. No. ZRB1409, Anti-POLR2A, clone 3G21 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit 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 NIH/3T3 cell lines were performed using a 1:100 dilution of Cat. No. ZRB1409, Anti-POLR2A, clone 3G21 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit 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 NIH/3T3 cell lines were performed using a 1:100 dilution of Cat. No. ZRB1409, Anti-POLR2A, clone 3G21 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit 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 HeLa (Lane 1), PC12 (Lane 2), MCF7 (Lane 3), and NIH3T3 (Lane 4) cell lysates were probed with Cat. No. ZRB1409, Anti-POLR2A, clone 3G21 ZooMAb® Rabbit Monoclonal (1:1,000 dilution). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates POLR2A (~220 kDa).
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 | rabbit |
| clone | 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 | IgG |
| mol wt | calculated mol wt 217.18 kDa |
| observed mol wt ~220 kDa | |
| packaging | pkg of 25 μL |
| product line | ZooMAb® learn more |
| recombinant | expressed in HEK 293 cells |
| shipped in | ambient |
| species reactivity | mouse, human, rat |
| storage temp. | 2-8°C |
| technique(s) | immunocytochemistry: suitable |
| immunohistochemistry: suitable | |
| western blot: suitable | |
| UniProt accession no. | P24928  |
| Application: | Anti-POLR2A, clone 3G21 ZooMAb, Cat. No. ZRB1409, is a recombinant Rabbit monoclonal antibody that targets DNA-directed RNA Polymerase II and is tested in Immunocytochemistry, Immunohistochemistry, and Western Blotting. |
| Application: | Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected POLR2A in HeLa, PC12, and NIH/3T3 cell lysates. Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected POLR2A in human testis tissue sections. Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected POLR2A in HeLa, A431, HUVEC and NIH 3T3 cell lines. 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: | Three repeats of YSPTSPS, phosphorylated at S5. |
| Legal Information: | ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany |
| Physical form: | Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS with 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: | 0.03 mg/mL after reconstitution at 25μL per vial. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type. |
| Specificity: | Clone 3G21 is a ZooMAb rabbit recombinant monoclonal antibody that specifically detects DNA-directed RNA polymerase II subunit A (POLR2A). It targets phosphorylated and non-phosphorylated YSPTSPS repeats in the C-terminal region. |
| Storage and Stability: | Recommended 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) is encoded by the POLR2A (also known as POLR2) gene (Gene ID: 5430) in human. (see MABE1793). It is the largest and 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 |
| Storage Temp. | 2-8°C |
| UNSPSC | 12352203 |