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Rat IgG2b Isotype Control Antibody

Recombinant Rat IgG2b Isotype Control, Rat IgG2b Negative Control Antibody, rtIgG2b Isotype Control

Catalog No. Product Name Size List Price (US$) Quantity
PA007144 In Vivo Grade Recombinant Rat IgG2b Isotype Control Antibody 1 mg 150.00
PA007144 In Vivo Grade Recombinant Rat IgG2b Isotype Control Antibody 5 mg 300.00
PA007144 In Vivo Grade Recombinant Rat IgG2b Isotype Control Antibody 25 mg 800.00
PA007144.05 In Vivo Grade Recombinant Rat IgG2b Isotype Control Antibody, Endotoxin 0.05 EU/mg 5 mg 525.00
PA007144.05 In Vivo Grade Recombinant Rat IgG2b Isotype Control Antibody, Endotoxin 0.05 EU/mg 25 mg 1350.00
Description

Rat IgG2b isotype control and mutants (natural and engineered rtIgG2b isotype controls) are useful for functional study using animal model among a number of in vitro and in vivo research applications. Rat IgG2b antibodies are more like mouse IgG2a and IgG2b, and human IgG1 antibodies, which are mostly used by the therapeutic antibody drugs. Most of rat antibodies are used to study mouse target proteins.

There are more and more IgG antibody formats with Fc engineered. The format with the Fc silenced, Fc silent, or Fcs with silenced effector function is the most popular. We provide Fc effector-silent rat IgG2b isotype controls.

Please contact us to ask for a quote for the engineered rat IgG2b isotype control mutants, rtIgG2b isotype control and mutants with Avi-, His-, and DYKDDDDK-tags, and biotinylated rtIgG2b isotype control and mutants. A variety of conjugates (such as dyes and fluorophores) with the rat IgG2b isotype control and Fc mutants are available.

PA007144: In vivo Grade Recombinant Rat IgG2b Isotype Control Antibody (rtIgG2b Isotype Control)

The in vivo grade recombinant rat IgG2b isotype control antibody (rtIgG2b isotype control) was produced in mammalian cells.
Immunogen: N/A.
Clone: 12B9.
Isotype: rat IgG2b, kappa.
Applications: an isotype-matched negative control for rat IgG2b 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 1x PBS.
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 rtIgG2b isotype control is 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 IgG1 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.

References of Rat IgG2b Isotype Control Antibody:


Isolation of rat IgM to IgG hybridoma isotype switch variants and analysis of the efficiency of rat Ig in complement activation
Pluschke, G., et al. Eur J Immunol. 1989 Jan;19(1):205-11. doi: 10.1002/eji.1830190121. PMID: 2646134
Sequential sublining was used in combination with enzyme-linked immunosorbent assays to isolate mu----gamma isotype switch variants of the rat IgM secreting mouse-rat B cell hybridoma line BA1.8. Switch variants to all four subclasses of IgG were obtained. Efficient lysis of sheep erythrocytes coated with the O18 antigen was observed with IgM and all IgG subclasses, but no lysis was triggered by IgE or IgA. One hundred to 1000 IgG molecules were required to mediate the same hemolytic activity as one IgM molecule. IgM and all IgG subclasses triggered C3 binding to the K1 encapsulated bacteria.
Tags: activity of Rat IgG2b Isotype Control; Rat IgG2b Isotype Control

Rat monoclonal antibodies to mouse IgG1, IgG2a, IgG2b, and IgG3 subclasses, and kappa chain isotypic determinants
Labit, C., et al. Hybridoma. 1984 Spring;3(1):163-75. doi: 10.1089/hyb.1984.3.163. PMID: 6207100
Rats of the LOU/Ws1 strain were immunized with mixtures of mouse monoclonal antibodies (MAbs) of various isotypes, and their spleen cells were fused with the rat myeloma Y3.Ag1.2.3 or the mouse myeloma X63.Ag8.653. From four fusion experiments, we have selected 14 rat MAbs that exhibited selective binding to either IgG2a, IgG2b, IgG1, and IgG3 subclasses or to kappa chain isotypic determinants. Cross-blocking studies revealed that three rat MAbs identified distinct determinants on the Fc fragment of the MAb H10-81.10 (A.TH, Igh-1e). By contrast, the IgG1, IgG2b, IgG3, and kappa isotypes defined by the mAbs analyzed in this study were found to be in close spatial relationship. These rat MAbs bound IgG2a, IgG2b, or IgG1 mouse MAbs, expressing the Igh-1e,j,a, or c, Igh-3a or b, or Igh-4a or b allelic specificities, respectively.
Tags: Rat IgG2b Isotype Control mAb; human IgG1 recombinant isotype control

The biological half-lives of four rat immunoglobulin isotypes
Peppard, J. V., et al. Immunology. 1980 Oct;41(2):405-8. doi: Not available. PMID: 7429545
Purified preparations of monoclonal rat IgG2a, IgG1, IgM and monomeric IgA, and of polyclonal IgG, were lightly radiolabelled with 125I and injected intravenously into rats. Blood samples were taken so that the half-life of each immunoglobulin isotype could be calculated. Half-life was defined as the time taken for the specific radioactivity present in the blood 24 h after injection to fall by half. The half-life values thus obtained were 106 h for IgG2a, 53 h for IgG1, 63 h for polyclonal IgG, 25 h for IgM and 27 h for monomeric IgA.
Tags: Rat IgG2b Isotype Control antibody for cancer research; Rat IgG2b Isotype Control antibody for mouse tumor model

Classes and subclasses of rat antibodies: reaction with the antigen and interaction of the complex with the complement system
Medgyesi, G. A., et al. Immunology. 1981 Aug;43(4):727-36. doi: Not available. PMID: 7251048
Antibodies against the dinitroaminophenyl (DNAP) haptenic group were raised in outbred CFY rats using HSA or LPS as carrier. Antibodies isolated by immunoadsorbent techniques were resolved into fractions representing distinct isotypes, and the resulting fractions were tested for avidity. Subclass IgG2a was found to contain antibodies of an avidity index lower than those of other IgG subclasses or IgM. IgG2a was the only isotype detected when DNAP-LPS was used for immunization. Complexes containing defined isotypes were compared for their capacity to activate homologous complement.
Tags: Rat IgG2b Isotype Control for cancer research; Rat IgG2b Isotype Control for mouse tumor model

Specificity of fc receptors for IgG2a, IgG1/IgG2b, and IgE on rat macrophages
Uede, T., et al. J Exp Med. 1981 Jul 1;154(1):374-89. doi: 10.1084/jem.154.1.374. PMID: 6167653
To characterize the Fc receptors on rat alveolar and peritoneal macrophages (M phi), we analyzed their ability to form rosettes with fixed ox erythrocytes (Eo') coated with myeloma proteins of all rat Ig classes and with fresh erythrocytes (Eo) sensitized with rat IgG1 and IgG2, rabbit IgG and IgM, and mouse IgA antibodies. The M phi formed rosettes with Eo' coated with rat myeloma proteins of classes IgG1, IgG2a, IgG2b, and IgE but not IgG2c, IgA, IgM, and IgD. Rat M phi also formed rosettes with Eo' coated with human IgG1, IgG3, IgG4, mouse IgG1, IgG2a, IgG2b, and rabbit IgG. Trypsin treatment of rat M phi abolished IgG1/IgG2b and IgE but not IgG2a rosettes. The results indicate that rat M phi have at least three distinct Fc receptors: one is specific for rat IgG2a and is trypsin resistant; a second is specific for rat IgE and is trypsin sensitive; and a third reacts with rat IgG1 rat IgG2b, and heterologous IgG and is trypsin sensitive.
Tags: in vivo study of Rat IgG2b Isotype Control antibody; bulk Rat IgG2b Isotype Control

For more references about Rat IgG2b Isotype Control Antibody, please contact our scientific support team with message@sydlabs.com.

Questions and Answers:

Question: What is an isotype control antibody and why is it used?
Answer: An ideal isotype control matches the isotype of the primary antibody used in the application but lacks binding to the target. It is normally used as a negative control to differentiate any non-specific background signal from the specific antibody signal. Different from our recombinant rat IgG2b isotype control (catalog number PA007144), other rat IgG2b kappa isotype controls against KLH, TNP, DNP,  GST, HEL, His-tag, DYKDDDDK, c-myc, HA, and GFP meet the requirements in some cases but do not in specific cases in the presence of the targets or cross-reactivity.

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 antibody. 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 mutations such as LALAPG 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 IgG2b isotype control (catalog # PA007144) 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.

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 IgG3 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 Mouse IgG3 Isotype Control Antibody
In vivo Grade Recombinant Rat IgG1 Isotype Control Antibody
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 IgG1 Fc Protein (hIgG1)
In vivo Grade Recombinant Human IgG2 Fc Protein (hIgG2)
In vivo Grade Recombinant Human IgG4 Fc Protein (hIgG4)
In vivo Grade Recombinant Mouse IgG1 Fc Protein (mIgG1)
In vivo Grade Recombinant Mouse IgG2a Fc Protein (mIgG2a)
In vivo Grade Recombinant Mouse IgG2b Fc Protein (mIgG2b)
In vivo Grade Recombinant Rat IgG2a Fc Protein (rtIgG2a)
In vivo Grade Recombinant Rat IgG2b Fc Protein (rtIgG2b)
In vivo Grade Recombinant Llama IgG2b Fc Protein (lIgG2b)
In vivo Grade Recombinant Rabbit IgG Fc Protein (rIgG)

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