I am treating tet on mice with doxycline (76ppm) for 2 weeks and i dont get the overexpression according to Western Blot. In rtPCR i see differences, but in protein level these differences are not reflected. What might be the issue?
Could be increased ubiquitination/degradation of the protein.
I am treating mice with doxycycline (75ppm) to induce gene expression but i see dermatitis in both control and transgenic mice.
12 March 2024 4,848 1 View
Hello! I am trying to detect TNFa in supernatans of THP1 macrophages (induced with 50ng/ml PMA) after stimulation with 0.1 ng/ml LPS for 24hrs. I am using Elisa but I haven’t managed to detect any...
25 January 2024 387 3 View
I am trying to detect TNFa in supernatant of AC16 cells using ELISA. To induce inflammation i incubate the cells with LPS (0.1 μg/ml) for 6 hrs. I havent managed to detect TNFa. Does anyone have a...
29 October 2023 3,842 1 View
I want to do a drug screening assay in a 96well plate and test protein level of cytokines with elisa.
02 August 2023 2,568 2 View
Hello, I am currently having problems with RNA extraction. I am using mouse liver (C57BL6J), and I have extracted RNA from mouse liver before. Before this experiment, my final RNA pellets were...
11 August 2024 7,082 3 View
I got these smeared bands quite often lately. We typically run the gel at 140V with a 10-12% gel and do a wet transfer at 220 mA for 1.5 hr in cold room. We also noticed some dirty spots/dots (see...
10 August 2024 7,480 3 View
Can anyone share any reference that gives mouse specific CD3 antibody sequence. I have been searching for this since few days and no luck. Thank you very much
08 August 2024 9,157 4 View
Hi all, I was just wondering if anyone has experience with multiplexing a mouse monoclonal primary and a rat primary. I'm trying to multiplex by incubating them in the same well but was told by a...
06 August 2024 9,710 1 View
i have sorted anti-NP specific plasma cells from bone marrow of C57BL/6 mice at certain times after immunization with variable counts and isolated total RNA using TRIZOL method for RT-PCR using...
05 August 2024 8,835 1 View
I have tried several times to isolate lymphocytes from mouse spleen, but all attempts have been unsuccessful. I tried most available protocols. I used different dissociation media (HBSS with Ca...
04 August 2024 9,913 7 View
I´ve been unable to find specific information about this neurotrophin in the CNS of rabbits exclusively. There is extensive info in mice, fish and rats, but in brain´s rabbit is hard to find....
04 August 2024 762 1 View
I need to lyse and homogenise mouse tissues. So far I have experimented with liver and lung tissue, using matrix D. The liver was completely lysed with 6 repetitions of the standard program, but...
31 July 2024 9,186 0 View
I need to Postfix mice brains. I slice the brains using microtome at 40 microns. These are Postnatal days 7, 11, 14, and 21. I am using half the brain for other analysis, so I need to fresh freeze...
31 July 2024 1,123 4 View
We have encountered an issue with the serum of mice being hemolyzed after the process of centrifugation. Could you please provide guidance to prevent hemolysis ?
29 July 2024 8,402 4 View