Anti-CD3d Antibody, clone 1H2 ZooMAb® Rabbit Monoclonal
SIGMA/ZRB1295 - recombinant, expressed in HEK 293 cells
Product Type: Chemical
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human spleen (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. ZRB1295, Anti-CD3d, clone 1H2 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous staining was observed in areas of red & white pulp of human spleen tissue lysate.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human spleen (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. ZRB1295, Anti-CD3d, clone 1H2 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous staining was observed in areas of red & white pulp of human spleen tissue lysate.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human spleen (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. ZRB1295, Anti-CD3d, clone 1H2 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous staining was observed in areas of red & white pulp of human spleen tissue lysate.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human tonsil (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. ZRB1295, Anti-CD3d, clone 1H2 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous staining was observed in both germinal center & non-germinal center cells of human tonsil tissue lysate.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human tonsil (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. ZRB1295, Anti-CD3d, clone 1H2 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous staining was observed in both germinal center & non-germinal center cells of human tonsil tissue lysate.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human tonsil (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. ZRB1295, Anti-CD3d, clone 1H2 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous staining was observed in both germinal center & non-germinal center cells of human tonsil tissue lysate.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of Jurkat cells was performed using a 1:100 dilution of Cat. No. ZRB1295, Anti-CD3d, clone 1H2 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 Jurkat cells was performed using a 1:100 dilution of Cat. No. ZRB1295, Anti-CD3d, clone 1H2 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 Jurkat cells was performed using a 1:100 dilution of Cat. No. ZRB1295, Anti-CD3d, clone 1H2 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 Jurkat (Lane 1), Hut78 (Lane 2), and MOLT-4 (Lane 3) cells were probed with Cat. No. ZRB1295, Anti-CD3d, clone 1H2 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 CD3d (~24 kDa).
Affinity Binding Assay Enhanced Validation - Recombinant Antibody Technology 100 mg/mL of non-specific control peptide (Run 1) or CD3d peptide (Run 2) were analyzed by affinity binding assay using Cat. No. ZRB1295, Anti-CD3d, clone 1H2 ZooMAb® Rabbit Monoclonal. The binding of this antibody to CD3d peptide displayed a KD of 1.0 x 10-12 whereas binding to non-specific control peptide displayed a KD of 4.4 x 10-3.
Flow Cytometry Enhanced Validation - Recombinant Antibody Technology Staining of one million Jurkat cells was performed using 1 μg of Cat. No. ZRB1295, Anti-CD3d, clone 1H2 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 | 1H2, recombinant monoclonal |
| conjugate | unconjugated |
| description | recombinant, expressed in HEK 293 cells |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation  |
| epitope sequence | Extracellular domain |
| 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 18.93 kDa |
| observed mol wt ~24 kDa | |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| Protein ID accession no. | NP_000723  |
| Quality Level | 200 |
| recombinant | expressed in HEK 293 cells |
| shipped in | ambient |
| species reactivity | human |
| species reactivity (predicted by homology) | monkey |
| 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. | P04234 |
| Application: | Quality Control Testing Evaluated by Western Blotting in Jurkat cell lysate. Western Blotting Analysis: A 1:1,000 dilution of this antibody detected CD3d in Jurkat cell lysate. Tested applications Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected CD3d in Hut78 and MOLT-4 cell lysates. Affinity Binding Assay: A representative lot of this antibody bound CD3d peptide with a KD of 1.0 x 10-12 in an affinity binding assay. Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected CD3d in Jurkat cells. Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected CD3d in human spleen and human tonsil tissue sections. Flow Cytometry Analysis: 1 μg from a representative lot detected CD3d in one million Jurkat cells. Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user |
| Application: | Anti-CD3d, clone 1H2 ZooMAb®, Cat. No. ZRB1295, is a recombinant Rabbit monoclonal antibody that specifically targets CD3d and is tested for use 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. Learn more about ZooMAb here. |
| Immunogen: | KLH-conjugated linear peptide corresponding to 15 amino acids from the extracellular domain of human T-cell surface glycoprotein CD3 delta chain (CD3d). |
| 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. 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 1H2 is a ZooMAb® rabbit recombinant monoclonal antibody that specifically detects human T-cell surface glycoprotein CD3 delta chain (CD3d). It targets an epitope within 15 amino acids from 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: | T-cell surface glycoprotein CD3 delta chain (UniProt: P04234; also known as T-cell receptor T3 delta chain, CD3d) is encoded by the CD3D (also known as T3D) gene (Gene ID: 915) in human. CD3d is a single-pass type I membrane protein that is part of the TCR-CD3 complex present on T-lymphocyte cell surface. It plays an essential role in adaptive immune response. When antigen presenting cells (APCs) activate T-cell receptor (TCR), TCR-mediated signals are transmitted across the cell membrane by the CD3 chains CD3D, CD3E, CD3G and CD3Z. All CD3 chains contain immunoreceptor tyrosine-based activation motifs (ITAMs) in their cytoplasmic domain. ITAM domain of CD3d is localized to amino acids 138-166. Upon TCR engagement, these motifs are phosphorylated by LCK and FYN resulting in the activation of downstream signaling pathways. CD3d is also reported to be involved in thymocyte differentiation and in the absence of a functional TCR-CD3 complex, thymocytes do not differentiate properly. CD3d also interacts with CD4 and CD8 to establish a functional link between the TCR and coreceptors CD4 and CD8, which is shown to be essential for activation and positive selection of CD4 or CD8 T-cells. CD3d is synthesized with a signal peptide (aa 1-21), which is subsequently cleaved off to generate the mature form that contains an extracellular domain (aa 106-126), a transmembrane domain (aa 106-126), and a cytoplasmic domain (aa 127-171). Mutations in CD3D gene are known to cause Immunodeficiency 19 that is characterized by onset in early infancy of recurrent bacterial, viral, and fungal infections. 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 |
| Storage Temp. | 2-8°C |