Hamster IgG1 Isotype Control Antibody

Both Armenian hamster IgG1 isotype control and Syrian hamster IgG1 isotype control (natural and engineered hamster IgG1 isotype controls) are useful for in vivo functional study using mouse and rat research model among a number of in vitro and in vivo research applications from ELISA, Western Blot (WB) to cell-based assays. The amino acid and DNA sequences of hamster IgG antibodies are different from the sequences of mouse and rat antibodies. Most of hamster antibodies are used to study mouse and rat target proteins. The ideal recombinant hamster IgG1 isotype control for in vivo use has low or no specific binding to any mouse and rat sample, and murinized or completely mouse variable regions in addition to the hamster IgG1 kappa constant regions.

There are more and more IgG antibody formats with Fc engineered. The formats with the Fc silenced, Fc silent, or Fcs with silent effector function is the most popular. Thus, we also provide Fc silent effector function hamster IgG isotype controls, including hamster IgG1 and IgG2 kappa isotype controls.

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

PA007146: In Vivo Grade Recombinant Armenian Hamster IgG1 Isotype Control Antibody

The in vivo grade recombinant Armenian hamster IgG1 isotype control antibody (hamster IgG isotype control) was produced in mammalian cells.
Immunogen: N/A.
Clone: 12B9.
Isotype: hamster IgG1, kappa.
Applications: an isotype-matched negative control for hamster 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 1x PBS.
Purity: >95% by SDS-PAGE under reducing conditions.
Endotoxin: Less than 1 EU/mg of protein as determined by LAL method.

PA007148: In Vivo Grade Recombinant Syrian Hamster IgG1 Isotype Control Antibody

The in vivo grade recombinant Syrian hamster IgG1 isotype control antibody (hamster IgG isotype control) was produced in mammalian cells.
Immunogen: N/A.
Clone: 12B9.
Isotype: hamster IgG1, kappa.
Applications: an isotype-matched negative control for hamster 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 1x PBS.
Purity: >95% by SDS-PAGE under reducing conditions.
Endotoxin: Less than 1 EU/mg of protein as determined by LAL method.

Shipping: The recombinant hamster IgG1 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

Hamster IgG antibodies are classified to three subclasses mainly based on the isotype of the heavy chain (IgG1, IgG2, and IgG3 for the heavy chain; kappa and lamda for the light chain), of which kappa is 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.

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

References of Hamster IgG1 Isotype Control Antibody:

Dynamic encounters with red blood cells trigger splenic marginal zone B cell retention and function.
Liu, D., et al. Nat Immunol. 2024 Jan;25(1):142-154. doi: 10.1038/s41590-023-01690-z. PMID: 38049580.
The antibodies used for staining were BV785-conjugated anti-B220 (RA3-6B2, 103246; 1:200), BV605-conjugated anti-CD19 (6D5, 115540; 1:200), Pacific Blue-conjugated anti-CD21/CD35 (7E9, 123414; 1:200), PE/Cy7-conjugated anti-CD23 (B3B4, 101614; 1:200), PE-conjugated anti-CD5 (53-7.3, 100608; 1:200), PerCP-Cy5.5-conjugated anti-IgM (RMM-1, 406512; 1:200), PE-conjugated anti-CD45.2 (104, 109808; 1:200), FITC-conjugated anti-CD45.1 (A20, 110706; 1:200), FITC-conjugated anti-CD1d (1B1, 123508; 1:200), APC-conjugated anti-TER-119 (TER-119, 116212; 1:200), FITC-conjugated anti-CD41 (MWReg30, 133903; 1:200), PE-conjugated anti-IgD (IA6-2, 348204; 1:200), Pacific Blue-conjugated anti-IgM (MHM-88, 314514; 1:200), AF700-conjugated anti-CD1c (L161, 331529; 1:200) and FITC-conjugated anti-CD27 (M-T271, 356404; 1:200) from Biolegend; APC hamster IgG1 isotype control (anti-TNP, A19-3, 553974; 1:200), AF647-conjugated anti-Ki-67 (B56, 558615; 1:200) and PE hamster IgG1 isotype control (anti-TNP, A19-3, 553972; 1:200) from BD Biosciences; PE-conjugated anti-CD55 (REA300, 130-104-023; 1:100), APC-conjugated anti-CD97 (REA678, 130-110-229; 1:100), Annexin V-FITC (130-093-060; 1:200), APC-conjugated anti-CD97 (REA1242, 130-124-980; 1:100) and APC-conjugated anti-CD55 (REA1231, 130-124-497; 1:100) from Miltenyi Biotec. ...Spleen marginal zone (MZ) B cells are important for antibody responses against blood-borne antigens. ...The signals they use to detect exposure to blood are not well defined. ...We show that MZ B cells use adhesion G-protein-coupled receptor ADGRE5 (CD97) for retention in the spleen. ...CD97 function in MZ B cells depends on its ability to undergo autoproteolytic cleavage and signaling via Gα13 and ARHGEF1. ...Red blood cell expression of the CD97 ligand CD55 is required for MZ B cell homeostasis.
Tags: activity of Hamster IgG1 isotype control; activity of Hamster IgG1 isotype control mAb

Reversing PD-1 Resistance in B16F10 Cells and Recovering Tumour Immunity Using a COX2 Inhibitor.
Pi, C., et al. Cancers (Basel). 2022 Aug;14(17):4134. doi: 10.3390/cancers14174134. PMID: 36077671.
The following antibodies were used as isotype controls: APC-Cy™7 Mouse IgG2a, κ Isotype Control (557751; BD Pharmingen), PE Hamster IgG1 κ Isotype Control (553972; BD Pharmingen), PerCP-Cy™5.5 Rat IgG2a, κ Isotype Control (550765; BD Pharmingen), FITC Rat IgG2a, κ Isotype Control (553929; BD Pharmingen), APC Mouse IgG2a, κ Isotype Control (551414; BD Pharmingen), FITC Rat IgM, κ Isotype Control (555951; BD Pharmingen), PE Rat IgG2b, κ Isotype Control (553989; BD Pharmingen). ...Immunotherapy is an effective method for tumour treatment. Anti-programmed cell death protein 1 (PD-1) and anti-programmed death-ligand 1 (PD-L1) monoclonal antibodies play a significant role in immunotherapy of most tumours; however, some patients develop drug resistance to PD-1/PD-L1 therapy. ...Cyclooxygenase-2 (COX2) is expressed in various solid tumours, and prostaglandin E2 (PGE2) drives the development of malignant tumours. ...We developed a drug-resistant B16F10 (B16F10-R) tumour mouse model through four rounds of selection in vivo. ...Additionally, we explored the role of COX2 in acquired resistance to pembrolizumab, an anti-PD-1 treatment. Immune cell infiltration was significantly decreased in resistant tumours compared to B16F10-NR tumours; however, ptgs2 gene expression was significantly elevated in resistant tumours.
Tags: Hamster IgG1 isotype control antibody; Hamster IgG1 recombinant isotype control

Rare t(X;14)(q28;q32) translocation reveals link between MTCP1 and chronic lymphocytic leukemia.
Walker, J. S., et al. Nat Commun. 2021 Nov;12(1):6338. doi: 10.1038/s41467-021-26400-x. PMID: 34732719.
Immunomodulatory assessment of Eµ-MTCP1 peripheral blood by flow cytometry was performed as follows: PE hamster anti-mouse CTLA4 , PE hamster IgG1 κ isotype control, BV421 rat anti-mouse CD5, FITC rat anti-mouse CD45, Alexa Fluor® 647 rat anti-mouse CD19, LIVE/DEAD™ fixable near-IR dead cell stain, rat anti-mouse CD16/CD32, BV510 hamster anti-mouse CD3e, BV650 rat anti-mouse CD11-b, BB515 rat anti-mouse CD19, BUV737 rat anti-mouse CD5, APC rat anti-mouse CD93, BUV395 rat anti-mouse CD45R/B220, BV786 rat anti-mouse IgM, PerCP-Cy5.5 rat anti-mouse CD21, BV711 Rat anti-mouse CD23, BV605 Rat anti-mouse IgD. ...Rare, recurrent balanced translocations occur in a variety of cancers but are often not functionally interrogated. Balanced translocations with the immunoglobulin heavy chain locus (IGH; 14q32) in chronic lymphocytic leukemia (CLL) are infrequent but have led to the discovery of pathogenic genes including CCND1, BCL2, and BCL3. Following identification of a t(X;14)(q28;q32) translocation that placed the mature T cell proliferation 1 gene (MTCP1) adjacent to the immunoglobulin locus in a CLL patient, we hypothesized that this gene may have previously unrecognized importance. Indeed, here we report overexpression of human MTCP1 restricted to the B cell compartment in mice produces a clonal CD5+/CD19+ leukemia recapitulating the major characteristics of human CLL and demonstrates favorable response to therapeutic intervention with ibrutinib. ...Along this principle, we have identified a CLL patient with a previously undescribed t(X;14)(q28;q32) translocation, which leads to co-localization of the mature T cell proliferation 1 (MTCP1) coding region with the IGH locus, a situation analogous to the translocation of BCL2. ...The Xq28 locus containing MTCP1 is bicistronic, encoding two distinct transcripts as a result of an ancient insertion event. ...The Xq28 locus containing MTCP1 is bicistronic, encoding two distinct transcripts as a result of an ancient insertion event.
Tags: Hamster IgG1 isotype control antibody for cancer research; Hamster IgG1 isotype control antibody for flow cytometry

CTLA-4 expression by B-1a B cells is essential for immune tolerance.
Yang, Y., et al. Nat Commun. 2021 Jan;12(1):525. doi: 10.1038/s41467-020-20874-x. PMID: 33483505.
Fluorochrome-conjugated antibodies include: anti-CD21-FITC (clone 7E9, Biolegend, cat#123408), anti-IAd-FITC(clone AMS-32.1, BD, cat#553547), anti-GL7-FITC (clone GL7, BD, cat553666), anti-IgMb-FITC (clone AF6-78, BD,cat#553520), anti-IgMa-BV785(clone DS-1, Biolegend, cat#743890), anti-CD43-PE (clone S11, Biolegend, cat#143206), anti-CD138-PE (clone 281-2, Biolegend, cat#142504), anti-CD267-APC (clone eBioBF10-3, eBioscience, cat#17-5942-82), anti-CD38-Alexa488 (clone 90, Biolegend, cat#102714), anti- H2-M-Alexa488 (clone 2E5A, BD, cat# 552405), anti-CD5-PE-Cy5 (clone 53-7.3, Biolegend, cat#100610), anti-CD19-PE-Cy5.5 (clone ID3, Invitrogen, cat#35-0193-82), anti-IgG1-PE-Cy7(clone RMG1-1, Biolegend, cat#406614), anti-ICOS-PE-Cy7(clone C398.4A, Biolegend, cat#313520), anti-CD150-PE-Cy7(clone TC150-12F12.2, Biolegend, cat#115913), anti-IgM-PE-Cy7(clone R6-60.2, BD,cat#552867), anti-B220-Alexa700(clone RA3-6B2, Biolegend, cat#103232), anti-CD3-Alexa700(clone 145-2C11, Biolegend, cat#100216), anti-IgM-APC (clone RMM1, Biolegend, cat#406509), anti-Bcl6-APC (clone 7D1, Biolegend, cat#648305), anti-Ki67-Alexa647 (clone 11F6, Biolegend, cat#151206), anti-CTLA-4-PE(clone UC10-4F10-11, BD, cat#553720), PE Hamster IgG1 isotype control (clone A19.3, BD, cat#553972), anti-CTLA-4-APC (clone UC10-4F10-11, BD, cat#564331) anti-Foxp3-APC (clone FJK-16S, eBioscience, cat#17-5773-80), anti-Foxp3-PE (clone FJK-16S, eBioscience, cat#12-5773-82), anti-IgD-APC-Cy7(clone 11-26c.2a, Biolegend, cat#405716), anti-CD4-APC-Cy7(clone GK1.5, BD,cat#552051), anti-CD23-biotin (clone, B3B4, Biolegend, cat#101604), anti-CD80-biotin (clone 16-10A, BD, cat#553767), anti-CD86-biotin (clone GL-1, Biolegend, cat#105004), anti-CD95-Qdot605 (clone SA367H8, Biolegend, cat#152612), anti-IgG1a-biotin (clone 10.9, BD, cat#553500), anti-IgG1b-biotin (clone B68.2, BD, cat#553533), anti-CXCR5-Brilliant BV605 (clone L138D7, Biolegend, cat#145513), anti-CD11b-PB (clone M1/70, Biolegend, cat#101224), anti-Gr-1-PB(clone RB6-8C5, Biolegend,ca#108430), anti-TCRαβ-PB(clone H57, Invitrogen,cat#HM3628), anti-CD11c-PB (clone N418, Biolegend, cat#117322), anti-CD3ε-PB (clone 145-2C11, Biolegend, cat#100334), anti-F4/80-PB (clone BM8, Biolegend, cat#123124), anti-CD279 (PD-1)-Brilliant violet 785 (clone 29F.1A12, Biolegend, cat#135225), anti-CD25-PE(clone 7D4, BD, cat#558642). LEAF purified anti-CD154 (clone MR1, Biolegend, cat#106508), anti-CD4 (clone GK1.5, Biolegend, cat#100442), IgG isotype control (clone SHG-1, Biolegend, cat#70647), anti-IgMb (clone AF6-78, customer produced by Biolegend). ...Here, we report that expression of this immune-regulator in mouse B-1a cells has a critical function in maintaining self-tolerance by regulating these early-developing B cells that express a repertoire enriched for auto-reactivity. ...This impaired immune homeostasis results from B-1a cell dysfunction upon loss of CTLA-4. ...Therefore, CTLA-4-deficient B-1a cells up-regulate epigenetic and transcriptional activation programs and show increased self-replenishment. ...These activated cells further internalize surface IgM, differentiate into antigen-presenting cells and, when reconstituted in normal IgH-allotype congenic recipient mice, induce GCs and Tfh cells expressing a highly selected repertoire.
Tags: Hamster IgG1 isotype control for animal model; Hamster IgG1 isotype control for mouse tumor model

A protocol for the culture and isolation of murine synovial fibroblasts.
Zhao, J., et al. Biomed Rep. 2016 Aug;5(2):171-175. doi: 10.3892/br.2016.708. PMID: 27446536.
Purified rat anti-mouse CD106 (cat. no. 553330), purified rat immunoglobulin G2a (IgG2a), κ isotype control (cat. no. 553927), FITC goat anti-rat Ig secondary antibody (cat. no. 554016), phycoerythrin (PE) hamster anti-mouse CD54 (cat. no. 553253), PE hamster IgG1 κ isotype control (cat. no. 553972), biotin rat anti-mouse CD90.2 (cat. no. 553011), biotin rat IgG2b, κ isotype control (cat. no. 553987) and PE-streptavidin (cat. no. 554061) were obtained from BD Pharmingen (Franklin Lakes, NJ, USA), vimentin rabbit monoclonal antibody (Alexa Flour 488 Conjugate; cat. no. 9854) was obtained from Cell Signaling Technology (Beverly, MA, USA). ...The culture of synovial fibroblasts (SFs) is one of the most effective tools for investigating the pathology and physiology of synovial tissues and should prove useful for identifying the importance of SFs in disease as well as for the development of novel therapeutic approaches for several chronic joint diseases, such as rheumatoid arthritis. ...However, thus far, a detailed protocol for the primary culture and isolation of murine SFs has not been established. ...Therefore, the present study describes an easy and convenient method for isolating and culturing SFs from C57BL/6 mice. ...This protocol can be divided into 4 stages: Isolation of synovial tissues, isolation of SFs, seeding of SFs for growth in culture and purity analysis of SFs using the four cell markers, vimentin, cluster of differentiation 90.2 (CD90.2; Thy‑1.2), intracellular adhesion molecule 1 (CD54) and vascular cell adhesion molecule 1 (CD106).
Tags: purified Hamster IgG1 isotype control; bulk Hamster IgG1 isotype control

For more references about Hamster IgG1 Isotype Control Antibody please contact our scientific support team with message@sydlabs.com.

Questions and Answers:

Question: Why do you use mouse variable regions in your recombinant hamster 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 hamster IgG antibodies are used to study mouse target proteins. Many scientists use hamster IgG isotype controls with hamster antibody variable regions assuming that they induce the same immune response in mice, if there is any, as the hamster antibodies of interest. However, two hamster variable regions including the one in the hamster 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 hamster variable regions in the hamster IgG antibodies to test. Thus, it is better to use the mouse variable region in the hasmter IgG isotype control. (We also have the hamster IgG isotype controls with the hamster IgG variable regions. In addition, we have the recombinant hamster Fc proteins as the hamster IgG isotype controls. Please contact us if you are interested.)

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 hamster IgG isotype controls. There is significant advantage for the in vitro method. First, we do not use mouse ascites for antibody production. Hamster hybridoma cell lines usually adapt poorly to in vitro conditions, and generate ascites in immunocompromised mice rather than hamsters. 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 hamster IgG isotype controls?
Answer: Our recombinant hamster IgG isotype controls including the hamster IgG1 isotype control (catalog # PA007146) are not from MOPC21 nor C1.18.4 though we also produce and sell the MOPC21 and C1.18.4 hamster IgG isotype controls. (Please contact us and ask for MOPC21 and C1.18.4 derived recombinant hamster 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 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)