Anti-MICA Antibody, clone 4F8 ZooMAb® Rabbit Monoclonal
SIGMA/ZRB1697 - recombinant, expressed in HEK 293 cells
Synonym: MHC class I polypeptide-related sequence A; MIC-A
Product Type: Chemical
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human pancreas (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. ZRB1697, Anti-MICA, clone 4F8 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Membranous/ECM staining was observed in pancreatic islets and acinar cells of human pancreas tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human pancreas (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. ZRB1697, Anti-MICA, clone 4F8 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Membranous/ECM staining was observed in pancreatic islets and acinar cells of human pancreas tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human pancreas (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. ZRB1697, Anti-MICA, clone 4F8 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Membranous/ECM staining was observed in pancreatic islets and acinar cells of human pancreas tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human kidney (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. ZRB1697, Anti-MICA, clone 4F8 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Membranous/ECM staining was observed in tubules and glomeruli of human kidney tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human kidney (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. ZRB1697, Anti-MICA, clone 4F8 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Membranous/ECM staining was observed in tubules and glomeruli of human kidney tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human kidney (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. ZRB1697, Anti-MICA, clone 4F8 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Membranous/ECM staining was observed in tubules and glomeruli of human kidney tissue sections.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of K562 cells was performed using a 1:100 dilution of Cat. No. ZRB1697, Anti-MICA, clone 4F8 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 cell membrane.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of K562 cells was performed using a 1:100 dilution of Cat. No. ZRB1697, Anti-MICA, clone 4F8 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 cell membrane.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of K562 cells was performed using a 1:100 dilution of Cat. No. ZRB1697, Anti-MICA, clone 4F8 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 cell membrane.
Western Blotting Enhanced Validation - Recombinant Antibody Technology Lysates from HeLa (Lane 1), L6 (Lane 2), and NIH3T3 (Lane 3) cells were probed with Cat. No. ZRB1697, Anti-MICA, clone 4F8 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 MICA (~58 kDa).
Affinity Binding Assay Enhanced Validation - Recombinant Antibody Technology 100 mg/mL of non-specific control peptide (Run 1) or MICA peptide (Run 2) were analyzed by affinity binding assay using Cat. No. ZRB1697, Anti-MICA, clone 4F8 ZooMAb® Rabbit Monoclonal. The binding of this antibody to MICA peptide displayed a KD of 1.5 x 10-6 whereas binding to non-specific control peptide displayed a KD of 1.9 x 10-3.
Flow Cytometry Enhanced Validation - Recombinant Antibody Technology Staining of one million K562 cells was performed using 1 μg of Cat. No. ZRB1697, Anti-MICA, clone 4F8 ZooMAb® Rabbit Monoclonal (Yellow histogram) or the equivalent amount of 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 |
| 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 42.92 kDa |
| observed mol wt ~58 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 | rat, mouse, 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. | Q29983  |
| Application: | Quality Control Testing Western Blotting Analysis: A 1:1,000 dilution of this antibody detected MICA in NIH3T3 cell lysate.Tested applicationsWestern Blotting Analysis: A 1:1,000 dilution from a representative lot detected MICA in HeLa and L6 cell lysates.Immunocytochemist |
| Application: | Anti-MICA, clone 4F8 ZooMAb, Cat. No. ZRB1697, is a recombinant Rabbit monoclonal antibody that targets in MICA and is tested for use in Affinity Binding Assay, Flow Cytometry, Immunocytochemistry, Immunohistochemistry (Paraffin), 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. Learn more about ZooMAb here. |
| Immunogen: | KLH-conjugated linear peptide corresponding to 17 amino acids from the extracellular domain of human MHC class I polypeptide-related sequence A (MICA). |
| 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: | 30 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 4F8 is a ZooMAb rabbit recombinant monoclonal antibody that detects MHC class I polypeptide-related sequence A (MICA). It targets an epitope within the extracellular domain. |
| 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: | MHC class I polypeptide-related sequence A (UniProt: Q29983; also known as MIC-A) is encoded by the MICA (also known as PERB11.1) gene (Gene ID: 100507436) in human. MIC-A is a single-pass type I membrane protein that act as stress-induced self-antigen that are recognized by gamma-delta T cells and serves as ligands for the KLRK1/NKG2D receptor. It is recognized by cytotoxic gamma-delta T cells expressing the TCR variable region V delta1. It is expressed on the cell surface in gastric epithelium, endothelial cells, and fibroblasts and in the cytoplasm in keratinocytes and monocytes. Its expression has also been reported in tumor cells. It is proteolytically cleaved and released from the cell surface of tumor cells which impairs KLRK1/NKG2D expression and T-cell activation. MIC-A can be induced by heat shock and by exposure to DNA damaging conditions. It is synthesized with a signal peptide (aa 1-23), which is cleaved off in the mature form. Their mature forms contain an extracellular domain (aa 24-307), a transmembrane domain (aa 308-328), and a cytoplasmic domain (aa 329-383). Two isoforms of MIC-A have been described that are produced by alternative splicing. This ZooMAb recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Groh, V., et al. (1999). Proc. Natl. Acad. Sci USA. 96 (12); 6879-6884). |
| WGK Germany | WGK 2 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | 2-8°C |
| UNSPSC | 12352203 |