Anti-PUMA Antibody, clone 1L16 ZooMAb® Rabbit Monoclonal
SIGMA/ZRB1600 - recombinant, expressed in HEK 293 cells
Synonym: Bcl-2-binding component 3;isoforms 1/2;JFY-1;p53 up-regulated modulator of apoptosis
Product Type: Chemical
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human small intestine (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. ZRB1600, Anti-PUMA, clone 1L16 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Punctate cytoplasmic staining was observed in glandular cells of human small intestine tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human small intestine (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. ZRB1600, Anti-PUMA, clone 1L16 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Punctate cytoplasmic staining was observed in glandular cells of human small intestine tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human small intestine (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. ZRB1600, Anti-PUMA, clone 1L16 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Punctate cytoplasmic staining was observed in glandular cells of human small intestine tissue sections.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of A431 cells was performed using a 1:1,000 dilution of Cat. No. ZRB1600, Anti-PUMA, clone 1L16 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 cytoplasm.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of A431 cells was performed using a 1:1,000 dilution of Cat. No. ZRB1600, Anti-PUMA, clone 1L16 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 cytoplasm.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of A431 cells was performed using a 1:1,000 dilution of Cat. No. ZRB1600, Anti-PUMA, clone 1L16 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 cytoplasm.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HeLa cells was performed using a 1:1,000 dilution of Cat. No. ZRB1600, Anti-PUMA, clone 1L16 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 cytoplasm.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HeLa cells was performed using a 1:1,000 dilution of Cat. No. ZRB1600, Anti-PUMA, clone 1L16 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 cytoplasm.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HeLa cells was performed using a 1:1,000 dilution of Cat. No. ZRB1600, Anti-PUMA, clone 1L16 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 cytoplasm.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HepG2 cells was performed using a 1:1,000 dilution of Cat. No. ZRB1600, Anti-PUMA, clone 1L16 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 cytoplasm.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HepG2 cells was performed using a 1:1,000 dilution of Cat. No. ZRB1600, Anti-PUMA, clone 1L16 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 cytoplasm.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HepG2 cells was performed using a 1:1,000 dilution of Cat. No. ZRB1600, Anti-PUMA, clone 1L16 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 cytoplasm.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of NIH3T3 cells was performed using a 1:1,000 dilution of Cat. No. ZRB1600, Anti-PUMA, clone 1L16 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 cytoplasm.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of NIH3T3 cells was performed using a 1:1,000 dilution of Cat. No. ZRB1600, Anti-PUMA, clone 1L16 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 cytoplasm.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of NIH3T3 cells was performed using a 1:1,000 dilution of Cat. No. ZRB1600, Anti-PUMA, clone 1L16 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 cytoplasm.
Western Blotting Enhanced Validation - Recombinant Antibody Technology Lysates from NIH3T3 (Lane 1), HepG2 (Lane 2), and L6 (Lane 3) cells were probed with Cat. No. ZRB1600, Anti-PUMA, clone 1L16 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 PUMA (~18 kDa).
Affinity Binding Assay Enhanced Validation - Recombinant Antibody Technology 100 mg/mL of non-specific control peptide (Run 1) or PUMA peptide (Run 2) were analyzed by affinity binding assay using Cat. No. ZRB1600, Anti-PUMA, clone 1L16 ZooMAb® Rabbit Monoclonal. The binding of this antibody to PUMA peptide displayed a KD of 1.0 x 10-12, whereas binding to non-specific control peptide displayed a KD of 1.6 x 10-5.
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 | 1L16, recombinant monoclonal |
| conjugate | unconjugated |
| description | recombinant, expressed in HEK 293 cells |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation  |
| epitope sequence | C-terminal |
| 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 20.53 kDa |
| observed mol wt ~18 kDa | |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| Protein ID accession no. | NP_055232  |
| purified by | using Protein A |
| Quality Level | 200 |
| recombinant | expressed in HEK 293 cells |
| shipped in | ambient |
| species reactivity | mouse, human, 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. | Q9BXH1 |
| Application: | Quality Control Testing Evaluated by Western Blotting in HepG2 cell lysate. Western Blotting Analysis: A 1:1,000 dilution of this antibody detected PUMA in HepG2 cell lysate. Tested applications Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected PUMA in NIH3T3 and L6 cell lysates. Affinity Binding Assay: A representative lot of this antibody bound PUMA peptide with a KD of 1.0 x 10-12 in an affinity binding assay. Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected PUMA in human small intestine tissue sections. Immunocytochemistry Analysis: A 1:1,000 dilution from a representative lot detected PUMA in A431, HeLa, HepG2, and NIH3T3 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. |
| Immunogen: | KLH-conjugated linear peptide corresponding to 17 amino acids from the C-terminal region of human Bcl-2-binding component 3 (PUMA). |
| Legal Information: | ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany |
| Physical form: | Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS, 5% Trehalose, normal appearance a coarse or translucent resin. The PBS/trehalose components in the ZooMAb formulation can have the appearance of a semi-solid (bead like gel) after lyophilization. This is a normal phenomenon. Please follow the recommended reconstitution procedure in the data sheet to dissolve the semi-solid, bead-like, gel-appearing material. The resulting antibody solution is completely stable and functional as proven by full functional testing. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 μL. |
| Reconstitution: | 300 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 1L16 is a ZooMAb® Rabbit recombinant monoclonal antibody that specifically detects Bcl-2-binding component 3 (PUMA). It targets an epitope within 17 amino acids from 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: | Bcl-2-binding component 3, isoforms 1/2 (UniProt: Q9BXH1; also known as JFY-1, p53 up-regulated modulator of apoptosis. PUMA) is encoded by the BBC3 (also known as PUMA) gene (Gene ID: 27113) in human. PUMA is a member of the BH3-only Bcl-2 family that is localized in the mitochondria and serves as a critical mediator of p53-dependent and -independent apoptosis induced by a wide variety of stress stimuli and growth factor/cytokine withdrawal. It is normally expressed at a very low level in cells, however, it is rapidly induced under stress conditions. PUMA transduces death signals primarily to the mitochondria, where it acts indirectly on Bax and/or Bak by relieving the inhibition imposed by anti-apoptotic members. It is reported to bind and antagonize all antiapoptotic members of the Bcl-2 family and induce mitochondrial dysfunction, cytochrome c release, and caspase activation. Its BH3 domain, localized to amino acids 137-151, is required for its interaction with Bcl-2 like proteins. The BH3 domains forms an amphipathic a-helical structure that allows it to bind to anti-apoptotic Bcl-2 family proteins. Its C-terminal region contains a hydrophobic domain that directs its mitochondrial localization and both the BH3 domain and mitochondrial localization are shown to be essential induction of apoptosis. PUMA is considered as an essential mediator of p53-dependent and p53-independent apoptosis. Inhibition of PUMA can lead to apoptosis deficiency and increases the risk for cancer development and therapeutic resistance. This ZooMAb® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Yu, J., and Zhang, L. (2008). Oncogene. 27 (suppl. 1); S71-S83; Yu, J., et al. (2001). Mol Cell. 7(3); 673-682). |
| WGK Germany | WGK 1 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | 2-8°C |