Anti-Rb Antibody, clone 2M15 ZooMAb® Rabbit Monoclonal
SIGMA/ZRB1014 - recombinant, expressed in HEK 293 cells
Synonym: Rb; Retinoblastoma-
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. ZRB1014, Anti-Rb, clone 2M15 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Nuclear staining was observed in both germinal center and non-germinal center cells, as well as in epithelium 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. ZRB1014, Anti-Rb, clone 2M15 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Nuclear staining was observed in both germinal center and non-germinal center cells, as well as in epithelium 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. ZRB1014, Anti-Rb, clone 2M15 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Nuclear staining was observed in both germinal center and non-germinal center cells, as well as in epithelium of human tonsil tissue sections.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of HeLa cells was performed using a 1:100 dilution of Cat. No. ZRB1014, Anti-Rb, clone 2M15 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 cells was performed using a 1:100 dilution of Cat. No. ZRB1014, Anti-Rb, clone 2M15 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 cells was performed using a 1:100 dilution of Cat. No. ZRB1014, Anti-Rb, clone 2M15 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 cells was performed using a 1:100 dilution of Cat. No. ZRB1014, Anti-Rb, clone 2M15 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 cells was performed using a 1:100 dilution of Cat. No. ZRB1014, Anti-Rb, clone 2M15 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 cells was performed using a 1:100 dilution of Cat. No. ZRB1014, Anti-Rb, clone 2M15 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 HepG2 cells was performed using a 1:100 dilution of Cat. No. ZRB1014, Anti-Rb, clone 2M15 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 HepG2 cells was performed using a 1:100 dilution of Cat. No. ZRB1014, Anti-Rb, clone 2M15 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 HepG2 cells was performed using a 1:100 dilution of Cat. No. ZRB1014, Anti-Rb, clone 2M15 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 NIH3T3 cells was performed using a 1:100 dilution of Cat. No. ZRB1014, Anti-Rb, clone 2M15 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 NIH3T3 cells was performed using a 1:100 dilution of Cat. No. ZRB1014, Anti-Rb, clone 2M15 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 NIH3T3 cells was performed using a 1:100 dilution of Cat. No. ZRB1014, Anti-Rb, clone 2M15 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), NIH3T3 (Lane 2), and MCF7 (Lane 3) cell lysates were probed with Cat. No. ZRB1014, Anti-Rb, clone 2M15 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 Rb (Retinoblastoma associated protein) (~120 kDa).
Affinity Binding Assay Enhanced Validation-Recombinant Antibody Technology 100 mg/mL of non-specific GST protein (Run 1) or recombinant retinoblastoma associated protein (Run 2) were analyzed by affinity binding assay using Cat. No. ZRB1014, Anti-Rb, clone 2M15 ZooMAb® Rabbit Monoclonal. The binding of this antibody to Rb protein displayed a KD value of 2.6 x 107 whereas its binding to non-specific protein displayed a KD of 2.8 x 103.
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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 | 2M15, 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 106.16 kDa |
| observed mol wt 120 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, mouse, rat |
| storage temp. | 2-8°C |
| technique(s) | affinity binding assay: suitable |
| immunocytochemistry: suitable | |
| immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable | |
| western blot: suitable | |
| UniProt accession no. | P06400  |
| Application: | Anti-Rb, clone 2M15 ZooMAb, Cat. No. ZRB1014, is a recombinant Rabbit monoclonal antibody that targets Retinoblastoma-associated protein and is tested Affinity Binding Assay, Immunocytochemistry, Immunohistochemistry (Paraffin), and Western Blotting, |
| Application: | Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected Rb (Retinoblastoma associated protein) in HeLa, and NIH3T3 cell lysates. Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected Rb (Retinoblastoma associated protein) in HeLa, A431, HepG2 and NIH 3T3 cells. Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected Rb (Retinoblastoma associated protein) in human tonsil tissue sections. Affinity Binding Assay: A representative lot of this antibody bound recombinant Rb (Retinoblastoma associated protein) with a KD of 2.6 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: | His-tagged recombinant fragment corresponding to 158 amino acids from the C-terminal region of human Retinoblastoma-associated protein (Rb). |
| 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: | 0.03 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 2M15 is a ZooMAb rabbit recombinant monoclonal antibody that specifically detects Retinoblastoma-associated protein (Rb). 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: | Retinoblastoma-associated protein (UniProt: P06400; also known as p105-Rb, p110-RB1, pRb, Rb, pp110) is encoded by the RB1 gene (Gene ID: 5925) in human. Retinoblastoma (Rb) proteins act as a negative regulator of cell cycle and controls the passage of cells from G1 to S phase. It I a key regulator of entry into cell division that acts as a tumor suppressor. It acts as a transcription repressor of E2F1 target genes. In its underphosphorylated, active state it interacts with E2F1 and represses its transcription activity, leading to cell cycle arrest. Its action is regulated by phosphorylation of several serine and threonine residues on the protein. Phosphorylated Rb protein sequesters transcription factors allowing the transition from G1 to S phase. It is phosphorylated by CDK6 and CDK4, and subsequently by CDK2 at serine 567 in G1, thereby releasing E2F1 which is then able to activate cell growth. It is dephosphorylated in the late M phase. Phosphorylation at threonine 821 and 826 promotes interaction between the C-terminal domain and the Pocket domain (aa 373-771), and thereby inhibits interactions with heterodimeric E2F/DP transcription factor complexes. Mutations in Rb1 gene are known to cause childhood retinoblastoma, a malignant tumor that arises from the nuclear layers of the retina. 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 |