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Rat IgG1 Isotype Control Antibody

Recombinant Rat IgG1 Isotype Control, Rat IgG1 Negative Control Antibody, rIgG1 Isotype Control

Catalog No. Product Name Size List Price (US$) Quantity
PA007142 In Vivo Grade Recombinant Rat IgG1 Isotype Control Antibody 1 mg 295.00
PA007142 In Vivo Grade Recombinant Rat IgG1 Isotype Control Antibody 5 mg 995.00
PA007142 In Vivo Grade Recombinant Rat IgG1 Isotype Control Antibody 25 mg 2950.00
Description

Rat IgG1 isotype control and mutants (natural and engineered rIgG1 isotype controls) are useful for functional study using animal model among a number of in vitro and in vivo research applications from ELISA, Western Blot (WB) to cell-based assays. Rat IgG1 and IgG2a antibodies are more like mouse IgG1 than rat IgG2b antibodies. Most of rat antibodies are used to study mouse target proteins.

Please contact us to ask for a quote for the engineered such as Fc silenced rat IgG1 isotype control mutants, rIgG1 isotype control and mutants with Avi-, His-, and Flag-tags, and biotinylated rIgG1 isotype control and mutants. A variety of conjugates (such as dyes and fluorophores) with the rat IgG1 isotype control and Fc silenced mutants are available.

PA007142: In Vivo Grade Recombinant Rat IgG1 Isotype Control Antibody (rIgG1 Isotype Control)

The in vivo grade recombinant rat IgG1 isotype control antibody (rIgG1 isotype control) was produced in CHO cells.
Immunogen: N/A.
Clone: 12B9.
Isotype: rat IgG1, kappa.
Applications: an isotype-matched negative control for rat IgG1 kappa antibodies used in ELISA, Western Blot (WB), Flow Cytometry (Flow), Immunoprecipitation (IP), Immunohistochemistry (Paraffin) (IHC (P)), Immunohistochemistry (Frozen) (IHC (F)), and in vivo animal model research.
Formulation: 0.2 μM filtered solution of PBS, pH 7.0.
Purity: >95% by SDS-PAGE under reducing conditions.
Endotoxin Level: Less than 1 EU/mg of protein as determined by LAL method.

Shipping: The recombinant rat IgG1 isotype control and mutants are shipped with ice pack. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
1 month from date of receipt, 2 to 8°C as supplied.
3 months from date of receipt, -20°C to -70°C as supplied.

Background

Rat IgG antibodies are classified to four subclasses mainly based on the isotype of the heavy chain (IgG1, IgG2a, IgG2b, and IgG2c for the heavy chain; kappa and lamda for the light chain), of which IgG2b and IgG2a, and kappa are the most abundantnamed in serum. IgG antibodies are large molecules of about 150 kDa composed of four peptide chains, two identical heavy chains of about 50 kDa and two identical light chains of about 25 kDa. The four peptides are arranged in a Y-shape tetramer by disulfide bonds formed between the two heavy chains and between a heavy chain and a light chain.

Rat IgG1 antibodies can be purified by the protein G beads, and relatively lower yield by the protein A beads.

Questions and Answers:

Question: Why do you use mouse variable regions in your recombinant rat IgG isotype controls?
Answer: The isotype control is normally used to differentiate any non-specific background signal from the specific antibody signal, including the immunogenicity if used in animals. Most in vivo functional grade rat IgG antibodies are used to study mouse target proteins. Many scientists use rat IgG isotype controls with rat antibody variable regions assuming that they induce the same immune response in mice, if there is any, as the rat antibodies of interest. However, two rat variable regions including the one in the rat IgG isotype control do not necessarily induce the same immune response in mice because in some cases, even one amino acid difference can make difference. Mouse variable regions can be used to show the potential immunogenicity induced by the rat variable regions in the rat IgG antibodies to test. Thus, it is better to use the mouse variable region in the rat IgG isotype control.

Question: What is the advantage of the recombinant antibody production method compared to the hybridoma method?
Answer: We use chemically defined medium for cell culturing to produce the recombinant rat IgG isotype controls. There is significant advantage for the in vitro method. First, we do not use mouse ascites for antibody production. Rat hybridoma cell lines usually adapt poorly to in vitro conditions, and generate ascites in immunocompromised mice rather than rats. Second, although antibodies can be produced by cell culturing from over 90% of hybridomas, there is a finite and significant failure rate. The recombinant method can be of help. Third, some companies and scientists add FBS into hybridoma cell culturing medium, which results in antibody contamination. There is no such contamination in production of recombinant antibodies. Fourth, we can easily switch the isotype with different antibody constant regions, and generate Fc mutations such as D265A which is used in therapeutic antibody research. It is better to use the same isotype control mutation. 

Question: Do you use the antibody sequences of MOPC21 (hybridoma clone number, isotype: mouse IgG1, kappa) or C1.18.4 (hybridoma clone number, isotype: mouse IgG2a, kappa) to develop your recombinant rat IgG isotype controls?
Answer: Our recombinant rat IgG isotype controls including the rat IgG1 isotype control (catalog # PA007142) are not from MOPC21 nor C1.18.4 though we also produce and sell the MOPC21 and C1.18.4 rat IgG isotype controls. Please contact us and ask for MOPC21 and C1.18.4 derived recombinant rat IgG kappa isotype controls if you are interested.

Question: What is the target of your recombinant rat IgG1 kappa isotype control antibody?
Answer: Similar to our other rat IgG isotype control antibodies, the recombinant rat IgG1 kappa isotype control antibody (catalog # PA007142) does not bind to any protein or chemical, even to those popular antigens for isotype control antibodies such as KLH, TNP, DNP,  GST, Hen Egg Lysozyme (HEL), His-tag, Flag, c-myc, HA, and GFP. PA007142 is an engineered rat IgG1 kappa antibody without antigen binding activity. We also produce and sell recombinant rat IgG1 kappa isotype controls against KLH, TNP, DNP,  GST, HEL, His-tag, Flag, c-myc, HA, and GFP. Please contact us and ask for them if you are interested.

Question: Is PA007142 your prefered rat IgG1 kappa isotype control. Correct?
Answer: Yes because it was engineered to lose the target binding activity. Some clients are concerned about the possibility that it may bind another target after the binding to one target becomes weak. But so far, we have not found that the rat IgG1 isotype control binds to any other target. In addition, its production yield is higher so that we can reduce the production cost and our selling prices.

Other in vivo grade Recombinant IgG Isotype Control Antibodies and Mutants:
In vivo Grade Recombinant Human IgG1 Isotype Control Antibody and Mutants
In vivo Grade Recombinant Human IgG2 Isotype Control Antibody
In vivo Grade Recombinant Human IgG4-S228P Isotype Control Antibody and Mutants
In vivo Grade Recombinant Mouse IgG1 Isotype Control Antibody and Mutants
In vivo Grade Recombinant Mouse IgG2a Isotype Control Antibody and Mutants
In vivo Grade Recombinant Mouse IgG2b Isotype Control Antibody and Mutants
In vivo Grade Recombinant Mouse IgG2c Isotype Control Antibody and Mutants
In vivo Grade Recombinant Rat IgG2a Isotype Control Antibody
In vivo Grade Recombinant Rat IgG2b Isotype Control Antibody
In vivo Grade Recombinant Rat IgG2c Isotype Control Antibody
In vivo Grade Recombinant Hamster IgG1 Isotype Control Antibody
In vivo Grade Recombinant Hamster IgG2 Isotype Control Antibody

In vivo Grade Recombinant IgG Fc Proteins:
In vivo Grade Recombinant Human Fc Protein (hIgG1)
In vivo Grade Recombinant Human Fc Protein (hIgG2)
In vivo Grade Recombinant Human Fc Protein (hIgG4)
In vivo Grade Recombinant Mouse Fc Protein (mIgG1)

Fc ELISA Kits and Reagents:
Human Fc ELISA Kit
Mouse Fc ELISA Kit
Human Fc ELISA Reagent Kit
Mouse Fc ELISA Reagent Kit

Protein A, Resin, Columns, and ELISA Kits:
Recombinant Protein A
Protein A Affinity Resin
Pre-packed Protein A Column
Protein A ELISA Kit

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