Specific Ab responses were measured by conventional ELISA. Briefly, ELISA plates were coated overnight at room temperature with FTAg (15 μg/ml). A serial dilution of the sera was carried out to determine the titer, which is defined as the inverse of the highest serum dilution factor giving an absorbance of >0.2. The Ab titers were determined using the following HRP-conjugated secondary Abs: rabbit anti-mouse IgG (H+L)–HRP, rabbit anti-mouse IgG1–HRP, and rabbit anti-mouse IgG2a–HRP (South-ernBiotech, Birmingham, AL; all with 1:1000 dilutions). SureBlue (KPL, Gaithersburg, MD) was used as a peroxidase substrate. After 15 min, the reaction was stopped by the addition of 100 μl 1 M H2SO4 and the absorbance was read at 450 nm.
Rabbit anti mouse igg h l hrp
Manufactured by Southern Biotech
Sourced in United States
Rabbit anti-mouse IgG (H+L)–HRP is a secondary antibody that binds to mouse immunoglobulin G (IgG) antibodies. The 'HRP' stands for Horseradish Peroxidase, which is an enzyme conjugated to the antibody. This antibody-enzyme conjugate can be used in various immunoassay techniques, such as Western blotting, ELISA, and immunohistochemistry, to detect and quantify mouse IgG in samples.
Automatically generated - may contain errors
Lab products found in correlation
Rabbit anti mouse igg1 hrp, by Southern Biotech (2 mentions)
Rabbit anti mouse igg2a hrp, by Southern Biotech (2 mentions)
Anti caspase 9, by Abcam (1 mentions)
Anti e cadherin, by Abcam (1 mentions)
Anti vimentin, by Abcam (1 mentions)
Immobilon western chemilum hrp substrate, by Merck Group (1 mentions)
Anti bax, by Abcam (1 mentions)
Anti caspase 3, by Abcam (1 mentions)
Goat anti rabbit igg h l hrp, by Southern Biotech (1 mentions)
Anti bcl 2, by Abcam (1 mentions)
Bca protein assay kit, by Thermo Fisher Scientific (1 mentions)
Goat anti rabbit igg h l mouse human ads hrp, by Southern Biotech (1 mentions)
4 protocols using rabbit anti mouse igg h l hrp
1
FTAg-Specific Antibody Titration
2
ELISA for Measuring IgG Antibodies
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The IgG specific Ab responses were measured by conventional ELISA method. Briefly, ELISA plates were coated overnight at room temperature with FTAg (80 μg/ml). A serial dilution of the sera was carried out to determine the titer, which is defined as the inverse of the highest serum dilution factor giving an absorbance of >0.2. Titers for the Abs were determined using the following HRP-conjugated secondary Abs: Rabbit anti-mouse IgG (H+L)–HRP, Rabbit anti-mouse IgG1-HRP, Rabbit anti-mouse IgG2a-HRP (Southern Biotech; all with 1:1000 dilutions). SureBlue (KPL) was used as a peroxidase substrate. After 15 min, the reaction was stopped by the addition of 100 μl 1M H2SO4, and the absorbance was read at 450 nm.
3
Protein Expression Analysis in SCC and HSC Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
SCC-15-NC, SCC-15–shACTN1, HSC-3-NC, and HSC-3-shACTN1 cells were harvested, and the total protein was isolated with RIPA buffer. After protein quantification using a BCA Protein Assay kit (Thermo Scientific Pierce, Rockford, IL, USA), total protein (30 µg) was loaded for SDS-PAGE electrophoresis and transferred onto a PVDF membrane. After being blocked with 5% (w/v) nonfat dried milk in tris buffered saline (TBS), membranes were incubated with diluted primary antibodies (anti-caspase 3, 1:5000; anti-caspase 9, 1:3000; anti-bax, 1:3000; anti-Bcl2, 1:2000; anti-E-cadherin, 1:500; anti-vimentin, 1:2000; anti-ACTN1, 1:1000; all purchases were made from Abcam, Cambridge, MA, USA). After washing with TBS containing 1‰ Tween (TBST) thrice for 5 min, membranes were incubated with secondary antibodies (Goat Anti-Rabbit IgG(H+L)-HRP, 1:20,000; Rabbit Anti-Mouse IgG(H+L)-HRP, 1:10,000; all purchased were made from Southern Biotech, Birmingham, AL, USA). After washing with TBST thrice for 5 min, membranes were incubated with Immobilon Western Chemilum HRP Substrate (Millipore Corporation, Billerica, MA, USA).
4
Protein Expression Analysis in Liver Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Approximately 200 mg of liver tissue was added to 2 mL of RIPA lysate and 20 μL of protease inhibitor. The ultrasonic homogenizer was used to homogenize the sample in an ice bath until no granules were detected. After centrifugation, the middle layer of the liquid was collected (the upper layer was oil, and the lower layer had a little precipitation), and centrifugation was repeated one time. For the BCA method and protein hyperthermia degeneration, the cells were transferred to a PVDF membrane and 5% skim milk powder at room temperature for 1.5 h. The membrane was immersed in the TLR4 antibody 1:500, NLRP3 antibody 1:500, ASC antibody 1:300, Caspase-1 antibody 1:500, or NF-κB p65 antibody 1:1000 at 4°C overnight. After washing the membrane, goat anti-rabbit IgG (H+L), mouse/human ads-HRP (1:20,000; Southern Biotech, 4050-05), or rabbit anti-mouse IgG (H+L)-HRP (1:10000; Southern Biotech, 6170-05) was incubated with the membrane for 1.5 h at room temperature. After washing the film, ECL luminescence solution was added, and the film was developed. The semiquantitative analysis of the bands was performed using software. The OD value of the target protein was divided by the OD value of the β-actin band as the final result.
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
Revolutionizing how scientists
search and build protocols!