Anti-Laminin-5 γ2 Antibody, clone D4B5 ZooMAb® Mouse Monoclonal
SIGMA/ZMS1059 - recombinant, expressed in HEK 293 cells
Product Type: Chemical
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human skin (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. ZMS1059, Anti-Laminin-5 γ2, clone D4B5 ZooMAb® Mouse Monoclonal. Reactivity was detected using a Rabbit Anti-Mouse IgG and HRP-DAB. Membranous staining was observed in basement membrane of human epithelium tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human skin (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. ZMS1059, Anti-Laminin-5 γ2, clone D4B5 ZooMAb® Mouse Monoclonal. Reactivity was detected using a Rabbit Anti-Mouse IgG and HRP-DAB. Membranous staining was observed in basement membrane of human epithelium tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human skin (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. ZMS1059, Anti-Laminin-5 γ2, clone D4B5 ZooMAb® Mouse Monoclonal. Reactivity was detected using a Rabbit Anti-Mouse IgG and HRP-DAB. Membranous staining was observed in basement membrane of human epithelium tissue sections.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of A431 cells was performed using a 1:100 dilution of Cat. No. ZMS1059, Anti-Laminin-5 γ2, clone D4B5 ZooMAb® Mouse Monoclonal and visualized with a Goat Anti-Mouse 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 1:100 dilution of Cat. No. ZMS1059, Anti-Laminin-5 γ2, clone D4B5 ZooMAb® Mouse Monoclonal and visualized with a Goat Anti-Mouse 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 1:100 dilution of Cat. No. ZMS1059, Anti-Laminin-5 γ2, clone D4B5 ZooMAb® Mouse Monoclonal and visualized with a Goat Anti-Mouse 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 Recombinant human Laminin 5 g2 was probed with Cat. No. ZMS1059, Anti-Laminin-5 g2, clone D4B5 ZooMAb® Mouse Monoclonal (1:10,000 dilution). Proteins were visualized using a Goat Anti-Mouse IgG1 secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates Laminin-5 2 (~130 kDa).
ELISA Enhanced Validation - Recombinant Antibody Technology 0.1 mg of recombinant human Laminin 5 g2 and GST protein were analyzed by ELISA using various dilutions (starting at 3 mg/mL) of Cat. No. ZMS1059, Anti-Laminin-5 γ2, clone D4B5 ZooMAb® Mouse Monoclonal. Alkaline phosphatase-conjugated Goat Anti-Mouse secondary antibody was used for detection.
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 | mouse |
| clone | D4B5, recombinant monoclonal |
| conjugate | unconjugated |
| description | recombinant, expressed in HEK 293 cells |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation  |
| epitope sequence | Internal |
| 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 | IgG1 |
| mol wt | calculated mol wt 130.98 kDa |
| observed mol wt ~130 kDa | |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| Protein ID accession no. | NP_005553.2  |
| purified by | using protein G |
| Quality Level | 200 |
| recombinant | expressed in HEK 293 cells |
| shipped in | ambient |
| species reactivity | human |
| storage temp. | 2-8°C |
| technique(s) | ELISA: suitable |
| immunocytochemistry: suitable | |
| immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable | |
| western blot: suitable | |
| UniProt accession no. | Q13753 |
| Application: | Quality Control Testing Evaluated by Western Blotting with recombinant Laminin 5 g2. Western Blotting Analysis: A 1:10,000 dilution of this antibody detected human recombinant Laminin 5 g2. Tested applications Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected Laminin-5 gamma2 in A431 Cell line. Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected Laminin-5 gamma2 in human skin tissue sections. ELISA Analysis: 300 μg/mL from a representative lot detected Laminin-5 gamma2 in Laminin-5 with control GST. Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user |
| Application: | Anti-Laminin-5 g2, clone D4B5 ZooMAb®, Cat. No. ZMS1059, is a recombinant Mouse monoclonal antibody that detects Laminin-5 (g2 chain) and is tested for use in ELISA, 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. |
| Immunogen: | GST-tagged recombinant fragment corresponding to 227 amino acids from the internal region of human. |
| Legal Information: | ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany |
| Physical form: | Purified recombinant mouse monoclonal antibody IgG, lyophilized in PBS with 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 D4B5 is a ZooMAb® Mouse recombinant monoclonal antibody that specifically detects Laminin-5 g2. It targets an epitope within the internal region and detects both isoforms. |
| 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: | Laminin subunit gamma-2 (UniProt: Q13753; also known as Cell-scattering factor 140 kDa subunit, CSF 140 kDa subunit, Epiligrin subunit gamma, Kalinin subunit gamma, Kalinin/nicein/epiligrin 100 kDa subunit, Ladsin 140 kDa subunit, Laminin B2t chain, Laminin-5 subunit gamma, Large adhesive scatter factor 140 kDa subunit, Nicein subunit gamma) is encoded by the LAMC2 (also known as LAMB2T, LAMNB2) gene (Gene ID: 3918) in human. Laminin is a major basement membrane glycoprotein consisting of a, b, g chains that are bound to each other by disulfide bonds. Over 15 different isoforms have been reported in the laminin family that are expressed in a tissue-specific manner and play different role in each tissue. g2, a subunit of Laminin-5 (also known as Laminin-332) is synthesized with a signal peptide (aa 1-21), which is subsequently cleaved off to generate the mature form. It is reported to be frequently over-expressed as a monomer or as the b3- g2 heterodimer in invasive cancers and is regarded as an invasion marker. It has 8 EGF-like domains. The cell adhesion and motility activities of Laminin-5 are reported to be regulated by the proteolytic cleavage of the short arm of Laminin g2 chain by BMP-1 or MT1-MMP. This cleavage releases a 45 kDa N-terminal proteolytic fragment (g2pf). The short arm and g2 pf can be further cleaved by serine proteases and by matrix metalloproteinases resulting in the release of small N-terminal fragments. g2pf‐like fragments have been reported to be present in conditioned media of cultured cancer cells and in sera from human cancer subjects. Laminin 5 synthesis in shown to be enhanced by inflammatory cytokines and growth factors that help in wound repair. This ZooMAb® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Miyazaki, K., et al. (2016). Cancer Sci. 107(12); 1909-1918; Sato, H., et al. (2014). Cancer Sci. 105(2); 168-175; Amano S., et al. (2004). Br. J. Dermatol. 151(5); 961-970). |
| WGK Germany | WGK 1 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | 2-8°C |