Recombinant Rat IgG Antibody Mixes

Normal rat serum and purified rat IgGs from normal rat serum are commonly used as non-specific passive blocking solution for antibody-based immuno-assays. Recombinant rat IgG antibody mixes, as a direct alternative to normal rat serum and even rat IgGs purified from rat serum or a non-specific passive blocking solution, can be used to eliminate interfering antibodies from immuno-assay systems using capture and detection antibodies. There is higher percentage of rat IgG1 or IgG2b in serum from different strains of rats but it may not be the best for all applications. So, recombinant rat IgG antibody mixes with different percentages of rat IgG1, IgG2a, IgG2b, and IgG2c were made to meet customers' requirements of various applications. A customized recombinant rat IgG antibody mix with different percentages of rat IgG1, IgG2a, IgG2b, and IgG2c can be provided.

Features of recombinant rat IgG antibody mixes compared with normal rat serum and serum-derived rat IgGs:

1) Animal free production. CHO cells rather than rats are used in antibody production.

2) Lot-to-lot consistency. CHO cells which are used for antibody drug manufacturing are used to manufacture the rat IgG antibodies in the antibody mixes. All production is under control and predictable.

3) As a targeted or passive antibody blocker to interfering antibodies like HARA (Human Anti-Rat Antibodies) and no binding to human and mouse tissues. In contrast, some serum-derived rat IgGs may bind to unknown targets in some human and mouse tissues.

4) Adjustable percentages of the antibodies in the antibody mixes. A customized recombinant rat IgG antibody mix with different percentages of rat IgG1, IgG2a, IgG2b, and IgG2c can be easily made according to customers' applications.

Recombinant rat IgG antibody mixes are the best normal rat serum alternative as blocking reagents, and serum-derived rat IgGs alternative.

PA007411: Recombinant Rat IgG Antibody Mix with more rtIgG2a and rtIgG2b

The recombinant rat IgG antibody mix is a mixture of recombinant anti-HEL rat IgG1, IgG2a, IgG2b, and IgG2c kappa antibodies which bind to neither human nor mouse samples. The percentage of rat IgG2a and IgG2b is much higher than the other antibody subclasses in the antibody mix. 
Clone: HyHEL-10.
Isotype: rat IgG1, IgG2a, IgG2b, and IgG2c kappa.
Specificity/Sensitivity: HEL-specific; neither human nor mouse samples.
Applications: ELISA, flow cytometry, immunoblotting, immunoprecipitation, immunocytochemistry, immunofluorescence, and isotype controls. 
Reactivity: None of Human, Mouse, and Rat.
Purification: The recombinant rat IgG antibodies were affinity-purified from supernatant of mammalian cells cultured in a chemically defined medium by affinity-chromatography using protein A and G.
Form of Antibody: 1x PBS.
Purity: >95% by SDS-PAGE under reducing conditions.

PA007412: Universal Recombinant Rat IgG Antibody Mix

The universal recombinant rat IgG antibody mix is a mixture of recombinant anti-HEL rat IgG1, IgG2a, IgG2b, and IgG2c kappa antibodies which bind to neither human nor mouse samples. The percentage of rat IgG1 and IgG2c are in the same range as combination of IgG2a and IgG2b. 
Clone: HyHEL-10.
Isotype: rat IgG1, IgG2a, IgG2b, and IgG2c kappa.
Specificity/Sensitivity: HEL-specific; neither human nor mouse samples.
Applications: ELISA, flow cytometry, immunoblotting, immunoprecipitation, immunocytochemistry, immunofluorescence, and isotype controls. 
Reactivity: None of Human, Mouse, and Rat.
Purification: The recombinant rat IgG antibodies were affinity-purified from supernatant of mammalian cells cultured in a chemically defined medium by affinity-chromatography using protein A and G.
Form of Antibody: 1x PBS.
Purity: >95% by SDS-PAGE under reducing conditions.

Shipping: The recombinant rat IgG antibody mix 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. For maximum recovery of the product, centrifuge the original vial after thawing and before removing the cap.
1 month from date of receipt, 2 to 8°C as supplied.
12 months from date of receipt, -20°C to -70°C as supplied.

Questions and Answers:

Question: Normal rat serum is often used for blocking or saturating immunoassays, especially those involving tissue samples such as immunohistochemistry (IHC). How much should recombinant antibody mixes be added to replace normal rat serum?
Answer: Our recombinant rat IgG antibody mixes can be used as normal rat serum alternative as passive blocking reagent, and more specifically, the purified rat IgGs alternative. It was said that there is 60 to 80 mg/ml protein in a normal rat serum sample, and 5 to 10 mg/ml IgG in a typical rat serum and plasma sample. It means that there is 6.3% to 16.7% IgGs in proteins from normal rat serum. One may calculate the amount of rat IgGs in a rat serum sample. One should also consider the percentage of various rat IgG isotypes in the rat serum sample vs recombinant rat IgG antibody mixes. Conditions of optimal rat IgG antibody mix performance should be determined experimentally by customers.

The most popular human Fc blocking reagents include:
Anti-human CD16 (clone 3G8), and CD32 (Clone IV.3) antibodies: for flow cytometry, immunohistochemistry (IHC), and immunoprecipitation (IP). They are highly specific for FcγRII (CD32) and FcγRIII (CD16), and reduce background staining; they may not block all Fc receptors. Clones 10.1, and 22 or H22 are the most widely used anti-human FcγRI (CD64) antibodies for Fc receptor blocking in flow cytometry and functional assays. It prevents non-specific Fc receptor binding on monocytes, macrophages, and dendritic cells, improving antibody specificity.
Purified human IgGs or mixes of human isotype controls: for general Fc blocking in IHC and immunofluorescence (IF). The purifed human IgGs from serum is cheap and easy to use but it is less specific, and may even introduce unwanted immunoglobulins. Syd Labs supply various human IgG isotype controls and recombinant human IgG antibody mixes.
Normal mouse serum, purified mouse IgG or isotype controls: for flow cytometry. It can serve as a control to block Fc receptors but it is non-specific for Fc receptors. The purifed mouse IgGs from serum is cheap and easy to use but it is less specific, and may even introduce unwanted immunoglobulins. Our recombinant mouse IgG antibody mixes are affordable; the ratio of various IgGs is optimized and adjustable without any unwanted immunoglobulins.
Bovine Serum Albumin (BSA) or FBS: for general blocking in Western Blot (WB) and IHC. They are readily available to reduce non-specific binding but less effective at Fc receptor blocking.
Commercial Fc blocking solutions: for flow cytometry, IHC, and functional assays. They are normally pre-optimized and highly effective but more expensive than DIY solutions. 

Best choice of human Fc blocking reagents based on your application are:
For flow cytometry: anti-human CD16 (Clone 3G8), CD32 (Clone IV.3), and CD64 (Clone 10.1, and 22 or H22).
For Immunohistochemistry (IHC) and Immunofluorescence (IF): Normal mouse serum, purified mouse or human IgGs or isotype controls or commercial Fc blocking kits. Our recombinant mouse IgG antibody mixes and recombinant human IgG isotype controls are affordable; the ratio of various IgGs is optimized and adjustable without any unwanted immunoglobulins.
For Western Blot and ELISA: BSA or FBS.
For functional assays (e.g., blocking Fc-mediated effects): Commercial Fc blocking reagents.

Syd Labs provides the following anti-human Fc blocking reagents:
Anti-human CD16 monoclonal antibody (Clone: 3G8)
Anti-human CD32 monoclonal antibody (Clone: IV.3)
Anti-human CD64 monoclonal antibody (Clone: 22)
Purified human IgG
Recombinant human IgG antibody mixes
Recombinant human IgG isotype control antibodies
Recombinant mouse IgG antibody mixes
Recombinant mouse IgG isotype control antibodies
Recombinant rat IgG antibody mixes
Recombinant rat IgG isotype control antibodies

Syd Labs provides the following anti-mouse Fc blocking reagents:
Anti-mouse CD16/CD32 monoclonal antibody (Clone: 2.4G2)
Recombinant mouse IgG antibody mixes
Recombinant mouse IgG isotype control antibodies
Recombinant rat IgG antibody mixes
Recombinant rat IgG isotype control antibodies