The largest database of trusted experimental protocols

6 protocols using anti ovalbumin igg1 mouse elisa kit

1

Ovalbumin-Induced Allergy Biomarkers

Check if the same lab product or an alternative is used in the 5 most similar protocols
NALF was centrifugalized to collect the supernatant. The blood was collected from the vena cava, and then serum was collected via centrifugation. Nasal tissues of sacrificed mice were stored at −70°C, and then homogenated and centrifuged to collect the supernatant. The anti-OVA Ig (immunoglobulin) E, anti-OVA IgG1 and anti-OVA IgG1 levels in serum, and IFN-γ, IL-12, IL-4, IL-5, and IL-13 levels in the NALF and nasal mucosa were measured through enzyme-linked immunosorbent assays (ELISA) using an anti-ovalbumin IgE (mouse) ELISA kit (500840, Cayman, MI, USA), anti-ovalbumin IgG1 (mouse) ELISA kit (500830, Cayman, MI, USA), mouse anti-ovalbumin (Gal d 2) IgG2a ELISA Kit (600-120-02A, Biolead, China), mouse IFN-γ ELISA kit (SEKM-0031, Solarbio, China), mouse IL-12p40 ELISA kit (SEKM-0012, Solarbio, China), mouse IL-13 ELISA kit (SEKM-0014, Solarbio, China), mouse IL-5 ELISA kit (SEKM-0006, Solarbio, China), and mouse IL-4 ELISA kit (SEKM-0005, Solarbio, China) according to the manufacturers’ instructions.
+ Open protocol
+ Expand
2

Measuring IgE and IgG1 Levels in Mice

Check if the same lab product or an alternative is used in the 5 most similar protocols
At one day before the injection of OVA/Al(OH)3, blood samples were collected from the tail vein. On the final day, whole blood was collected by cardiac puncture. The blood samples were incubated at room temperature for 1 h at 4 °C overnight and were centrifuged at 10,000 rpm for 10 min to obtain the serum. The amount of total IgE, OVA-specific IgE and IgG1 was measured in serum by using BD OptEIA Mouse IgE ELISA Set (BD Bioscience, San Jose, CA, USA), DS Mouse IgE ELISA (OVA) (DS Pharma Biomedical, Osaka, Japan), Anti-Ovalbumin IgG1 (mouse) ELISA kit (Cayman Chemical, Ann Arbor, MI, USA), respectively.
+ Open protocol
+ Expand
3

Immunization and Antibody Response Profiling

Check if the same lab product or an alternative is used in the 5 most similar protocols
Control and NHA mice were immunized with albumin from chicken egg white (OVA, Ovalbumin) from Sigma (Grade VII). Each mouse received 25 µg of OVA with Imject™ Alum Adjuvant (ALUM, Thermo Fisher Scientific) at a ratio of 1:1 in the final volume of 100 µl per mouse administered i.p. After 14 days, mice were challenged once again with the same dose of OVA-ALUM. NHA mice were divided into three groups. NHA before mice were phlebotomized before first immunization, NHA boost mice were phlebotomized before second OVA immunization, and NHA both were phlebotomized before first and second immunization (see Supplementary Fig. 3c). Untreated mice received Imject™ Alum Adjuvant without OVA. Blood was obtained from mice 14 days after the second immunization; plasma was isolated and stored at −80 °C. The concentration of anti-OVA IgG antibodies was determined using Anti-Ovalbumin IgG1 (mouse) ELISA Kit (Cayman Chemical).
+ Open protocol
+ Expand
4

IgG1 Quantification from Mouse Serum

Check if the same lab product or an alternative is used in the 5 most similar protocols
Blood was collected by cardiac puncture at endpoints and serum isolated by coagulation and centrifugation. Serum aliquots were snap-frozen at −80° C until use. Total IgG1 was measured using IgG (Total) Mouse ELISA Kit (Thermo) and OVA-specific IgG1 was measured using Anti-Ovalbumin IgG1 (mouse) ELISA Kit (Cayman Chemical) on duplicate sera samples diluted 1:10,000 and 1:100 respectively. Absorbance values within dynamic range were read at 450 nm and 570 nm, and final concentrations were interpolated on Sigmoidal (4PL) fit of standard-curve.
+ Open protocol
+ Expand
5

Measurement of Proinflammatory Cytokines by ELISA

Check if the same lab product or an alternative is used in the 5 most similar protocols
Proinflammatory cytokines (TNF-α, IL-1β and IL-6) serum concentrations were measured using Enzyme-linked immunosorbent assay (ELISA) kits (ThermoFisher). Briefly, ELISA plates coated with cytokine capture antibody were washed, blocked, and incubated with 50 μl of serum samples for 2 hours. In parallel, a standard curve was made by incubating 2-fold serial dilutions of the standards. Plates were then washed and incubated with biotin-conjugated detection antibodies for 1 hour. After washing, plates were incubated with Streptavidin-HRP. Bound antibodies were detected by adding 3,3′,5,5′-tetramethylbenzidine substrate (TMB Solution) followed by plate reading for absorbance at 450 nm. Cytokines levels were calculated based on a standard curve. OVA-specific IgG1 was measured by using the anti-Ovalbumin IgG1 (mouse) ELISA Kit (#500830, CaymanChemical) according to the manufacturer’s instructions.
+ Open protocol
+ Expand
6

Quantification of Anti-OVA Antibodies

Check if the same lab product or an alternative is used in the 5 most similar protocols
Blood samples were collected from the tail vein of mice on day 28, and anti-OVA antibodies were detected using Anti-Ovalbumin IgG1 (mouse) ELISA Kit (Cayman Chemical, MI, United States). The antibody levels were measured according to the manufacturer’s instructions using Infinite F200 Pro microplate reader (Tecan, Grödig, Austria).
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!