Anti-PCNA Antibody, clone 3B22 ZooMAb® Rabbit Monoclonal
SIGMA/ZRB1442 - recombinant, expressed in HEK 293 cells
Synonym: Proliferating cell nuclear antigen
Product Type: Chemical
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human tonsil (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. ZRB1442, Anti-PCNA, clone 3B22 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Nuclear staining was primarily observed in germinal center cells, with a lesser frequency of staining observed in non-germinal center cells of human tonsil tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human tonsil (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. ZRB1442, Anti-PCNA, clone 3B22 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Nuclear staining was primarily observed in germinal center cells, with a lesser frequency of staining observed in non-germinal center cells of human tonsil tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human tonsil (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. ZRB1442, Anti-PCNA, clone 3B22 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Nuclear staining was primarily observed in germinal center cells, with a lesser frequency of staining observed in non-germinal center cells of human tonsil tissue sections.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of HeLa cell line was performed using a 1:100 dilution of Cat. No. ZRB1442, Anti-PCNA, clone 3B22 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 line was performed using a 1:100 dilution of Cat. No. ZRB1442, Anti-PCNA, clone 3B22 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 line was performed using a 1:100 dilution of Cat. No. ZRB1442, Anti-PCNA, clone 3B22 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 line was performed using a 1:100 dilution of Cat. No. ZRB1442, Anti-PCNA, clone 3B22 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 line was performed using a 1:100 dilution of Cat. No. ZRB1442, Anti-PCNA, clone 3B22 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 line was performed using a 1:100 dilution of Cat. No. ZRB1442, Anti-PCNA, clone 3B22 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 line was performed using a 1:100 dilution of Cat. No. ZRB1442, Anti-PCNA, clone 3B22 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 line was performed using a 1:100 dilution of Cat. No. ZRB1442, Anti-PCNA, clone 3B22 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 line was performed using a 1:100 dilution of Cat. No. ZRB1442, Anti-PCNA, clone 3B22 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) and HepG2 (Lane 2) cell lysates were probed with Cat. No. ZRB1442, Anti-PCNA, clone 3B22 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 PCNA (~34 kDa).
Affinity Binding Assay Enhanced Validation-Recombinant Antibody Technology 100 mg/mL of non-specific protein (Run 1) or recombinant PCNA protein (Run 2) were analyzed by affinity binding assay using Cat. No. ZRB1442, Anti-PCNA, clone 3B22 ZooMAb® Rabbit Monoclonal. The binding of this antibody to PCNA displayed a KD value of 2.0 x 107 whereas its binding to non-specific protein displayed a KD of 1.8 x 104.
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 (recombinant) |
| clone | 3B22, 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 | IgG |
| mol wt | calculated mol wt 28.77 kDa |
| observed mol wt 34 kDa | |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| recombinant | expressed in HEK 293 cells |
| shipped in | ambient |
| species reactivity | human |
| storage temp. | 2-8°C |
| technique(s) | affinity binding assay: suitable |
| immunocytochemistry: suitable | |
| immunohistochemistry: suitable | |
| western blot: suitable | |
| UniProt accession no. | P48454  |
| Application: | Anti-PCNA, clone 3B22 ZooMAb, Cat. No. ZRB1442, is a recombinant Rabbit monoclonal antibody that specifically targets PCNA and is tested for in Affinity Binding Assay, Immunocytochemistry, Immunohistochemistry, and Western Blotting. |
| Application: | Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected PCNA in HeLa cell lysate. Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected PCNA in HeLa, A431, HUVEC cell lines. Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected PCNA in human tonsil tissue sections. Affinity Binding Assay: A representative lot of this antibody bound recombinant PCNA with a KD of 2.0 x 10-7 in an affinity binding assay. 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: | GST/His-tagged recombinant fragment corresponding to the first 150 amino acids from the N-terminal region of human Proliferating cell nuclear antigen (PCNA). |
| Legal Information: | ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany |
| Physical form: | Purified recombinant rabbit monoclonal antibody IgG, lyophilized in lyophilized from 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: | 0.3 mg/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 3B22 is a ZooMAb rabbit recombinant monoclonal antibody that specifically detects Proliferating cell nuclear antigen (PCNA). It targets an epitope within the first 150 amino acids. |
| 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: | Serine/threonine-protein phosphatase 2B catalytic subunit gamma isoform (UniProt: P48454; also known as EC:3.1.3.16, CAM-PRP catalytic subunit, Calcineurin, testis-specific catalytic subunit, Calmodulin-dependent calcineurin A subunit gamma isoform, PPP3CC) is encoded by the PPP3CC (also known as CALNA3, CNA3) gene (Gene ID: 5533) in human. PPP3CC is a calcium-dependent, calmodulin stimulated protein phosphatase that plays an essential role in the transduction of intracellular Ca2+-mediated signals. It is activated by Ca2+-bound calmodulin following an increase in intracellular Ca2+. At low Ca2+ concentrations, the catalytic subunit (aa 52-343; calcineurin A) is inactive and is bound to the regulatory subunit (calcineurin B) in which only two high-affinity binding sites are occupied by Ca2+. In response to elevated calcium levels, the occupancy of the low-affinity sites on calcineurin B by Ca2+ causes a conformational change of the C-terminal regulatory domain of calcineurin A, resulting in the exposure of the calmodulin-binding domain (aa 386-400) and in the partial activation of calcineurin A. The subsequent binding of Ca2+-bound calmodulin leads to the displacement of the autoinhibitory domain (aa 451-489) from the active site and possibly of the autoinhibitory segment (aa 401-408) from the substrate binding site which fully activates calcineurin A. This ZooMAb recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. |
| WGK Germany | WGK 1 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | 2-8°C |
| UNSPSC | 12352203 |