Anti-AFP Antibody, clone 3B7 ZooMAb® Rabbit Monoclonal
SIGMA/ZRB1159 - recombinant, expressed in HEK 293 cells
Product Type: Chemical
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Frozen section human fetal liver (A) and Negative control (no primary) (B) tissue sections were prepared using standard methods. Immunostaining was performed using a 1:1,000 dilution of Cat. No. ZRB1159, Anti-AFP, clone 3B7 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit and HRP-DAB. Membranous/ECM staining was observed in human fetal liver tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Frozen section human fetal liver (A) and Negative control (no primary) (B) tissue sections were prepared using standard methods. Immunostaining was performed using a 1:1,000 dilution of Cat. No. ZRB1159, Anti-AFP, clone 3B7 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit and HRP-DAB. Membranous/ECM staining was observed in human fetal liver tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Frozen section human fetal liver (A) and Negative control (no primary) (B) tissue sections were prepared using standard methods. Immunostaining was performed using a 1:1,000 dilution of Cat. No. ZRB1159, Anti-AFP, clone 3B7 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit and HRP-DAB. Membranous/ECM staining was observed in human fetal liver tissue sections.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HepG2 cell line was performed using a 1:100 dilution of Cat. No. ZRB1159, Anti-AFP, clone 3B7 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488. 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 cell line was performed using a 1:100 dilution of Cat. No. ZRB1159, Anti-AFP, clone 3B7 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488. 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 cell line was performed using a 1:100 dilution of Cat. No. ZRB1159, Anti-AFP, clone 3B7 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488. 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 HepG2 (Lane 1) and Human Fetal Liver (Lane 2) lysates were probed with Cat. No. ZRB1159, Anti-AFP, clone 3B7 ZooMAb® Rabbit Monoclonal (1:10,000 dilution). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates AFP (~60 kDa).
Affinity Binding Assay Enhanced Validation - Recombinant Antibody Technology 10 mg/mL of non-specific GST protein (Run 1) or recombinant AFP protein fragment (Run 2) were analyzed by affinity binding assay using Cat. No. ZRB1159, Anti-AFP, clone 3B7 ZooMAb® Rabbit Monoclonal. The binding of this antibody to AFP protein fragment displayed a KD of 1.9 x 10-9, whereas binding to non-specific GST protein displayed a KD of 6.2 x 10-4.
Flow Cytometry Enhanced Validation - Recombinant Antibody Technology Staining of one million HepG2 was performed using 1.0 μg of Cat. No. ZRB1159, Anti-AFP, clone 3B7 ZooMAb® Rabbit Monoclonal (Yellow histogram) or the equivalent amount of a Rabbit IgG isotype control (Grey histogram), followed by a PE-conjugated Donkey Anti-Rabbit IgG 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 | rabbit |
| clone | 3B7, 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 ~68.68 kDa |
| observed mol wt ~60 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 |
| storage temp. | 2-8°C |
| technique(s) | affinity binding assay: suitable |
| flow cytometry: suitable | |
| immunocytochemistry: suitable | |
| immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable | |
| western blot: suitable | |
| UniProt accession no. | P02771  |
| Application: | Quality Control Testing Evaluated by Western Blotting in Human Fetal Liver tissue lysate. Western Blotting Analysis: A 1:10,000 dilution of this antibody detected AFP in Human Fetal Liver tissue lysate. Tested ApplicationsWestern Blotting Analysis: A 1:10,000 dilution from a representative lot detected AFP in HepG2 cell lysate. Flow Cytometry Analysis: 1.0 μg from a representative lot detected AFP in HepG2 cells. Immunohistochemistry Applications: A 1:1,000 dilution from a representative lot detected AFP in human fetal liver tissue sections. Affinity Binding Assay Analysis: A representative lot of this antibody bound AFP proteins with a KD of 1.9 x 10^-9 in an affinity binding assay. Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected AFP in HepG2 cell line. Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user |
| Application: | Anti-AFP, clone 3B7 ZooMAb®, Cat. No. ZRB1159, is a recombinant Rabbit monoclonal antibody that specifically targets a-Fetoprotein and is used in Affinity Binding Assay, Flow Cytometry, Immunocytochemistry, Immunohistochemistry, and Western Blotting. |
| 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: | His-tagged recombinant fragment corresponding to 192 amino acids from the Albumin 1 domain, N-terminal region of human Alpha-fetoprotein. |
| 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: | 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 3B7 is a ZooMAb® rabbit recombinant monoclonal antibody that specifically detects alpha-fetoprotein. It targets an epitope within the Albumin 1 domain from the N-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: | Alpha-fetoprotein (UniProt: P02771; also known as Alpha-1-fetoprotein, Alpha-fetoglobulin, AFP) is encoded by the AFP (also known as HPAFP) gene (Gene ID: 174) in human. AFP is a secreted plasma protein that is found in fetuses older than 4 weeks and it reaches its highest levels around 12-16 weeks of gestation and then declines significantly after birth. AFP is synthesized in fetal liver with a signal peptide (aa 1-18), which is subsequently cleaved off to generate the mature form. It resembles albumin in many of its physicochemical properties and is involved in a variety of biological functions, including transporting metal ions, drugs, bilirubin, and steroids. Although in normal adults AFP levels are low, but it is reported to be a reliable marker of liver damage and its higher levels are associated with hepatitis, liver cirrhosis, and various cancers, including hepatocarcinoma, pancreatic cancer, and cancers of the reproductive system. It also serves as a reliable marker for testing for prenatal abnormalities. This ZooMAb® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Wang, X., and Wang, Q. (2018). Can. J. Gastroentrol. Hepatol. 2018; Article 9049252). |
| WGK Germany | WGK 1 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | 2-8°C |