Rat IgG_2b Isotype Control [Clone 1-2] — Purified in vivo PLATINUM™ Functional Grade

Clone 1-2 is most commonly used as a rat IgG2b isotype control in murine in vivo experiments, rather than as a function-blocking or depleting antibody. Its primary application is to serve as a negative control when testing other rat IgG2b monoclonal antibodies in mice.

Essential context and supporting details:

• Isotype controls like clone 1-2 are important for distinguishing non-specific binding from specific antibody-mediated effects in immunological assays, such as flow cytometry, immunohistochemistry, and in vivo antibody administration.
• Clone 1-2 does not neutralize cytokines or deplete cell populations; it is not used for targeting mouse proteins or mediating biological effects. It is administered in parallel with test antibodies in mouse studies to ensure that observed effects are due to specific antigen-antibody interactions and not simply the presence of rat IgG2b immunoglobulin.
• Its usage is particularly common in immunotherapy, tumor, and immune cell depletion studies where one needs to control for Fc-receptor binding, complement activation, or general antibody-mediated immunomodulatory effects. In such studies, clone 1-2 helps rule out artifacts or background caused by antibody isotype alone.

Additional relevant information:

• When used as a control, it is generally administered in the same dose and via the same route as the experimental antibody to provide a valid comparison.
• Experimental reports and vendor documentation confirm that there are no unique or therapeutic biological effects reported for clone 1-2 itself in mice aside from its role as a negative control reagent.

In summary, clone 1-2’s main in vivo application in mice is as a rat IgG2b isotype control, ensuring experimental rigor when using other monoclonal antibodies of the same isotype.

When considering antibodies commonly used alongside those targeting 1. Tumour Necrosis Factor (TNF) and 2. Programmed Death-1 (PD-1), several notable examples are mentioned in the literature:

Commonly Used Antibodies

1. Rituximab: A monoclonal antibody used for treating lymphoma and autoimmune diseases.
2. Infliximab: A chimeric monoclonal antibody used for rheumatoid arthritis treatment.
3. Nivolumab: A human monoclonal antibody used for treating malignant melanoma.
4. Panitumumab: A human monoclonal antibody targeting the Epidermal Growth Factor Receptor (EGFR) in metastatic colorectal cancer.
5. Daclizumab: A humanized monoclonal antibody used for preventing allograft rejection.

Proteins Used in Conjunction with Antibodies

1. CD14, CD20, CD34: Proteins targeted by peptide antibodies in research applications such as ELISA, ICC, WB, and FC.
2. Isocitrate Dehydrogenase, B-Raf, EGFR: Proteins quantified using peptide antibodies in diagnostics via IHC.
3. Epitope-specific peptides: Used in the study of antibody binding and cross-reaction, such as those derived from the hemagglutinin (HA) antigen of the H1N1 influenza virus.

Secondary Antibodies

While not proteins used in conjunction with primary antibodies in the same context, secondary antibodies are crucial for detecting bound primary antibodies:

• Anti-rabbit IgG, anti-mouse IgG: Common secondary antibodies used in indirect detection methods.

These antibodies and proteins are extensively used in research, diagnostics, and therapeutic applications, often in conjunction with TNF and PD-1 antibodies.

The key findings from scientific literature on clone 1-2 citations focus on two main areas: (1) studies of cloning methods and efficiencies, and (2) the phenomenon of 'cloned journals' and issues around scientific citations.

1. Cloning Methods and Efficiencies:

• High-efficiency Cloning Vectors: Advances in cloning vector design, such as the ‘two in one’ cloning vector based on pUC19, enable efficient one-step digestion-ligation and direct visual screening using a red fluorescent protein reporter. This method allows both blunt-end and T-A cloning with high transformation and verification rates, making it accessible to standard laboratories.
• Cloning Success Rates: Quantitative assessments of cloning efficiency (e.g., for lentiviral vectors) report that the total percentage of positive clones can approach 48% ± 7.6%, with monomeric insertion rates (single copy of target sequence per vector) close to 90% for several constructs.

2. Cloned Journals and Citations:

• Proliferation and Motivation: Cloned journals (unauthorized replicas of reputable publications) attract authors mainly due to open access at low cost and academic promotion prospects. Surveys indicate substantial agreement (average 79%) among writers regarding diverse motivations for submitting to cloned journals.
• Awareness of Consequences: Authors publishing in cloned journals are well aware of negative consequences—including potentially misleading academic credit and hampering scientific progress (average agreement of 79% for awareness of consequences).
• Threat to Integrity: Clone journals threaten the integrity of scientific publishing, undermining peer review and research validity. Publications in clone journals may propagate incorrect conclusions and negatively influence scientific progress.
• Citation Dynamics: Publications that fail replication tend to be cited more than those that are replicable—a counterintuitive finding that poses a risk to scientific reliability.

Additional Relevant Context:

• Fabricated Citations: Studies have found high proportions (47–69%) of fabricated citations in scientific literature, varying by discipline, especially geography and medicine.

In summary, scientific literature reveals both technical advances in cloning methodology (with efficient screening and high success rates) and serious issues in publication ethics caused by cloned journals and unreliable citation practices.

Dosing regimens for clone 1-2 antibodies in mouse models can vary significantly depending on several experimental factors, though specific standardized protocols are not well-established in the literature for this particular isotype control.

Key Factors Influencing Dosing Variations

The dosing regimens for clone 1-2 are primarily influenced by the mouse strain being used, the specific disease model being studied, and the experimental objectives. As a rat IgG2b isotype control, clone 1-2 serves as a negative control antibody rather than having therapeutic targeting properties. Its dosing must be matched to the experimental antibody being tested to ensure proper control conditions.

General Considerations for Isotype Control Dosing

Since clone 1-2 functions as an isotype control, its dosing should mirror that of the test antibody in the same experiment. This means that the amount, route of administration, and timing of clone 1-2 administration should match the experimental antibody to provide valid negative control data. The antibody is designed to have minimal cross-reactivity with mouse cells and tissues.

Lack of Standardized Protocols

Unlike therapeutic antibodies such as checkpoint inhibitors (which have well-defined dosing ranges like 200-500 μg per mouse for anti-PD-1 antibodies), clone 1-2 does not have established independent dosing guidelines. This is because it is not used to deplete specific cell types or neutralize cytokines but solely serves as a control. Researchers must determine appropriate dosing based on the specific requirements of their experimental system and the dosing of the primary antibody being investigated.

The variability in dosing regimens ultimately reflects the diverse applications and experimental designs across different mouse models, making it essential for researchers to optimize dosing parameters for their specific experimental context.