Sample
serum, plasma, hybridoma cell supernatants, ascites, other biological fluids
Detection Method
Sandwich-ELISA
Intended Use
This Mouse Immunoglobulin G (lgG) antigen assay is intended for the quantitative determination of total mouse lgG antigen in serum, plasma, hybridoma cell supernatants, ascites or other biological fluids. This assay does not distinguish lgG subclasses. For research use only.
Contents of Kit
1;Antibody coated microtiter strip plate;96T;4°C
2;10× Wash buffer;50ml;4°C
3; 5× Diluent;50ml;4°C
4;Mouse lgG standard (lyophilized);1 vial;4°C
5;Anti-mouse horseradish peroxidase antibody: 1 vial;4°C
6;TMB substrate solution;10ml;4°C
Storage
Store all kit components at 4°C upon arrival. Return any unused microplate strips to the plate pouch with desiccant. Reconstituted standard may be stored at -80°C for later use. Do not freeze-thaw the standard more than once. Store all other unused kit components at 4°C. This kit should not be used beyond the expiration date.
Precision
Intra-assay Precision: Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
Inter-assay Precision: Inter-assay Precislon: Three samples of known concentration were tested in ten Independent assays to assess inter-assay precislon.
Detection Range
0.5-500 ng/mL
Sensitivity
The minimum detectable dose (MDD) was determined by adding two standard deviations to the mean optical density value of twenty zero standard replicates(range OD450: 0.041-0.047) and calculating the corresponding concentration. The MDD was 0.12 ng/ml.
General Description
IgG is the most abundant immunoglobulin in serum and is predominately involved in the secondary immune response. The IgG subclasses are designated 1, 2, 3 and 4 based on their relative prevalence in human serum.
Standard Curve
Typical standard curve (EXAMPLE ONLY):
Citations
Publication ()
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Highly selective HPK1 inhibitor NDI-101150 mediates immune cell activation and robust antitumor responses, distinct from immune checkpoint blockade
Ciccone, D. N., Kuo, F. S., Boiko, S., Carreiro, S., Kaila, N., McElwee, J. J., Tummino, P. J., & Loh, C.
J Immunother Cancer2025 Jul 30PubMed ID: 40738502Read Article
Applications: ELISA
Reactive species: Mouse
"Abstract: Background: HPK1 (hematopoietic progenitor kinase 1, MAP4K1), an MAP4K serine/threonine kinase family member, is a negative regulator of immune cell function. Genetic HPK1 kinase inactivation or knockout in mice leads to immune cell activation and tumor growth suppression, providing a strong rationale for targeting HPK1 kinase activity as an immuno-oncology therapy.
Methods: A structure-guided drug design approach was used to identify a highly potent and selective small molecule inhibitor of HPK1 kinase activity, NDI-101150. The effect of NDI-101150 in vitro was evaluated on multiple immune cell types and in multiple syngeneic tumor models in vivo.
Results: Treatment with NDI-101150 enhanced T-cell activation in immune-suppressive or exhausted conditions, augmented B-cell activation, and upregulated dendritic cell function, including in settings where anti-programmed cell death protein-1 (PD-1) did not have an effect. These effects translated into significant inhibition of tumor growth in several syngeneic tumor models, including models that are less responsive to anti-PD-1. Importantly, NDI-101150 alone or in combination with anti-PD-1 mediated a highly effective antitumor response on tumor re-challenge, indicating that NDI-101150 induces immune memory against the tumor.
Conclusion: The data presented here indicate that a small molecule kinase inhibitor of HPK1 results in broad immune cell activation, leading to an efficacious antitumor immune response."
Article snippet: At day 14, NDI-101150 treatment not only resulted in a significant enhancement of KLH-IgG (Total IgG ELISA Kit, Creative Diagnostics, Cat. #DEIA8704), more so than anti-PD-1, but also augmented T-independent DNP-IgG, whereas anti-PD-1 had no effect. 
Figure 1. Quantification of KLH-specific IgM and IgG isotypes in blood from mice.
COA
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