Catalog No.	PA007411
Product Name	Rat IgG Antibody Mix with more rtIgG2a and rtIgG2b (Clone: HyHEL-10) | PA007411
Supplier Name	Syd Labs, Inc.
Brand Name	Syd Labs
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 with more rtIgG2a and rtIgG2b 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.
Note	Recombinant rat IgG antibody mixes: animal free production; lot-to-lot consistency; no binding to human or mouse tissues; adjustable formulation of the rat IgG antibodies in the antibody mixes. Best normal rat serum alternative; normal rat serum blocking reagent alternative; serum-derived rat IgGs alternative.
Order Offline	Phone: 1-617-401-8149 Fax: 1-617-606-5019 Email: message@sydlabs.com Or leave a message with a formal purchase order (PO) Or credit card.

Description

PA007411: Recombinant Rat IgG Antibody Mix with more rtIgG2a and rtIgG2b (Clone: HyHEL-10)

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.

Background of Recombinant Rat IgG Antibody Mix with more rtIgG2a and rtIgG2b

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. So, recombinant rat IgG antibody mixes with different percentages of rat IgG1, IgG2a, IgG2b, and IgG2c were made to meet customers’ requirements of different 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.

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 for blocking reagent, and more specifically, the purified normal 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 in 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): for flow cytometry, immunohistochemistry (IHC), and immunoprecipitation (IP). It is highly specific for FcγRII (CD32) and FcγRIII (CD16), and reduces background staining; it may not block all Fc receptors. Clones 10.1, and 22 or H22 are the most widely used anti-human FcγRI (CD64) clone for Fc 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). It 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.
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. It 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.

References of Recombinant Rat IgG Antibody Mixes:

1、Platelet activation contributes to hypoxia-induced inflammation.
Delaney C, et al. Am J Physiol Lung Cell Mol Physiol. 2021 Mar 1;320(3):L413-L421. doi: 10.1152/ajplung.00519.2020. PMID: 33264579.
“Thrombocytopenic mice were generated with an anti-GP1bα rat IgG antibody. …We evaluated male and female 9-wk-old normoxic and hypoxic mice and in selected experiments included hypoxic thrombocytopenic mice. …Platelet depletion prevents hypoxia-driven increases in the proinflammatory chemokines CXCL4 and CCL5 and attenuates hypoxia-induced increase in plasma CSF-2. …Platelets are small, anucleated cells derived from megakaryocytes, best known for their role in hemostasis. …Platelet granules store numerous growth factors, vasoactive mediators, chemokines, cytokines, and angiogenic agents, many of which have been implicated in vascular remodeling and thrombosis in PH.”

2、Development of a Novel Anti-Mouse CCR2 Monoclonal Antibody (C2Mab-6) by N-Terminal Peptide Immunization.
Tanaka, T., et al. Monoclon Antib Immunodiagn Immunother. 2022 Apr;41(2):80-86. doi: 10.1089/mab.2021.0063. PMID: 35377242.
“The secondary Alexa Fluor 488-conjugated anti-rat IgG and Alexa Fluor 488-conjugated anti-rabbit IgG were purchased from Cell Signaling Technology, Inc. (Danvers, MA). …Afterward, cells were treated with Alexa Fluor 488-conjugated anti-rat IgG (1:1000) or Alexa Fluor 488-conjugated anti-rabbit IgG (1:1000) as well, following the collection of fluorescence data, using a SA3800 Cell Analyzer (Sony Corp., Tokyo, Japan). …Then, cells were treated with Alexa Fluor 488-conjugated anti-rat IgG. …However, since the subclass of C2Mab-6 was rat IgG1, it did not possess ADCC and CDC. Therefore, in further studies, we will convert the subclass of C2Mab-6 into mouse IgG2a to evaluate whether the depletion of CCR2-expressing immunosuppressive cells can enhance antitumor activities in mouse syngeneic and xenograft models of human cancers. …It has been reported that the development of therapeutic agents targeting the CCL2-CCR2 axis can be effective for treating or managing various diseases.”

3、Simultaneous serodetection of major rat infectious pathogens by a multiplex immunochromatographic assay.
Tosa N, et al. Exp Anim. 2021 May 13;70(2):161-168. doi: 10.1538/expanim.20-0099. PMID: 33177250.
“For this assay, an ICA strip was placed into a microtube containing 150 µl PBS and either 0.75 µl of rat serum or 1.5 µl of whole blood. Binding antibodies were visualized by using anti-rat IgG antibody-conjugated colloidal gold. …Rapid and simple serologic tests that require only a small amount of blood without the euthanization of animals are valuable for microbial control in colonies of laboratory animals. …Under these conditions, the multiplex ICA simultaneously and specifically detected antibodies to multiple antigens. …The sensitivities of the multiplex ICA for Sendai virus, hantavirus, and sialodacryoadenitis virus were 100%, 100%, and 81%, respectively. …These observations indicate high specificity of the multiplex ICA.”

4、Pharmacokinetics of Nivolumab and Erythropoietin in a Rat Model of Diet-Induced Obesity.
Yi-Hua Sheng, et al. Pharm Res. 2025 Feb;42(2):271-280. doi: 10.1007/s11095-025-03819-1. PMID: 39849218.
“Endogenous rat IgG was also measured in the nivolumab study. A standard noncompartmental analysis was performed to calculate pharmacokinetic parameters. …Endogenous rat IgG was measured using an Abcam rat IgG ELISA kit following the manufacturer’s instructions. …In the nivolumab study, endogenous rat IgG in lean and obese cohorts was measured at the age of 15 weeks (before the nivolumab injection) and at 21 weeks (at the end of the study), as shown in Fig. …The concentration of rat IgG was statistically higher in obese rats at both times. …Biologics has been developed rapidly in the past decades, yet there is a lack of knowledge on biodisposition in special populations for many of them.”

5、Establishment of Novel Anti-Mouse CCR3 Monoclonal Antibodies (C3Mab-6 and C3Mab-7) by N-terminal Peptide Immunization.
Asano, T., et al. Monoclon Antib Immunodiagn Immunother. 2022 Apr;41(2):94-100. doi: 10.1089/mab.2021.0065. PMID: 35471054.
“A secondary Alexa Fluor 488-conjugated anti-rat IgG was purchased from Cell Signaling Technology, Inc., (Danvers, MA) as well. …Then, cells were treated with Alexa Fluor 488-conjugated anti-rat IgG (1:2000; Cell Signaling Technology, Inc.), after which fluorescence data were collected, using the BD FACSLyric (BD Biosciences, Franklin Lakes, NJ). …GPCRs expressed on the plasma membrane have native conformations. Therefore, immunization with cells, which express target GPCRs, is a suitable approach against GPCRs, such as chemokine receptors. …The Alexa Fluor 488-conjugated anti-rat IgG was subsequently added, after which fluorescence data were obtained, using BD FACSLyric. …In a previous study, we developed other anti-mCCR3 mAbs; C3Mab-2 and C3Mab-3, using the CBIS method, and showed their usefulness in flow cytometry.”

For more references about Recombinant Rat IgG Antibody Mixes please contact our scientific support team with message@sydlabs.com.

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

Rat IgG Antibody Mix with more rtIgG2a and rtIgG2b (HyHEL-10) from: Recombinant Rat IgG Antibody Mix with more rtIgG2a and rtIgG2b: PA007411 Syd Labs

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