What is the observed molecular weight of Immediate early gene Egr-1 in mouse cortex whole cell lysates?

Hi everyone, I used anti-Egr-1 (rabbit monoclonal, ab133695, Abcam) in western blotting experiment and I see multiple bands ranging from 50kDa to 130kDa, where at 130kDa bands are more intense...

02 March 2020 3,058 1 View

What is the difference between microRNA and siRNA ?

Hi everyone, Even though the definitions and descriptions are available online about microRNA ans siRNA, I want to get an insight through the scientists working on it and could give me a...

06 July 2018 5,325 1 View

If anyone can suggest me the source of caspase-8 antibody best working for mouse brain tissue?

Hi, I have to do IHC staining of caspase-8 in AD tg mouse brain tissue. If anyone can provide the best source and protocol for IHC-P staining of mouse brain tissue with caspase-8, it will be...

04 May 2018 6,038 1 View

Can anyone provide the protocol for IHC-P for the immunostaining with Cleaved Caspase-3 (Asp175) Antibody #9661?

With reference to the question, if anyone can share the reference paper or protocol for IHC- P staining and depicting the staining pattern in brain tissue for cleaved caspase 3. Thanks,

04 May 2018 7,857 0 View

Does immunostaining of total caspase 3 indicate the activation of caspase pathway for apoptosis?

31 December 2017 4,724 3 View

Does unilateral entorhinal cortex lesion lead to changes in contralateral hippocampi??

I came across some literature showing the evidences of equally altered gene expression of some markers in contralateral site in stroke and EC Lesion. In my experiment, from imuunohistochemistry...

11 December 2017 3,919 1 View

Can anyone provide the protocol for Acridine orange/Ethidium bromide staining to visualize apoptosis?

Hi everyone, Here I am working on neurodegeneration model and I am in a need to visualize apoptosis in early as well as late phase. I am aware of TUNEL assay, but I am more interested in...

08 September 2017 2,146 4 View

How to use cell counting plugin in Image J?

I need to count nerve cells in the tissue stained with Cresyl violet. Please provide me guide to make us of Image J software for cell counting. I have cell counter plugin in Image, but I don't...

07 August 2017 3,143 1 View

Why protein blocking step is needed before the application of primary antibody in IHC?

I am curious to know the possible reasons of protein blocking step and how does it affect the final result if we skip the step. Thanks in advance!

07 August 2017 4,724 0 View

If I am using microwave oven for antigen retrieval step in IHC, what could be the ideal exposure time?

Hi, Before I used to use waterbath for antigen retrieval step (citrate buffer) for IHC, but now I am advised to use microwave. I performed IHC 2-3 times, but I observe partial staining or no...

04 May 2017 6,901 10 View

How can I measure MMP-1 protein activity or protein level from liver tissue?

Instead of checking only the mRNA level, I want to check the active protein level of MMP-1 in Liver tissue from mice. How can I do that?

03 March 2021 1,763 2 View

IHC on mouse liver hepatocytes?

Can anyone help me choose primary antibodies for mouse liver hepatocytes and also in conjunction Cd8 cells. Also Cd3 cells. I want to look at hepatocytes and adjacent immune cells in mice with...

01 March 2021 6,953 1 View

How to avoid ulcers in s.c. injected tumors ?

Recently, we have injected s.c. B16F10 tumor cells in BL/6 mice. At around 12-15th day some animals showed ulcerations in tumors , with a big hole in the middle (it looks like a volcano). We have...

01 March 2021 9,938 3 View

Can someone suggest a way to prepare slides to observe under confocal microscope for observing fluorescent tagged proteins??

I am growing the cells on coverslips and transfecting the fluorescent tagged protein directly on them. Then i want to observe these cells under confocal microscope. Currently i am just using...

01 March 2021 6,142 3 View

Interested to stain mice tissue with WGA 488 conjugate, anybody has a protocol to recommend?

Interested to stain mice brain tissue with WGA 488 conjugate, appreciate if anyone willing to offer me some guidance or suggestions! Thank you!

28 February 2021 3,951 2 View

B-mercaptoethanol (BME) protocol?

Hi everybody, I have been running a multiplex IHC which allow me to stain the same tissue with different markers (Primary Ab + Secondary conjugated Ab) (AF487, AF555, AF647). Since I would like to...

27 February 2021 5,028 2 View

Why do my MCFs not form monolayer?

Hello, I am looking for some expertise on culturing MCF7s. I purchased them from ATCC and I have been following all media and culture conditions. Media is EMEM with 10% FBS and 10ug/mL of...

27 February 2021 3,904 2 View

Is there anyone who experienced with cytotoxic effect of Hoescht 33342 dye?

Is Hoechst toxic to cells? In general, it is known as non-toxic. I am working with dsRED MDA-MB-231 cell line. I applied it for my PDMS-based microfluidic platform, for this reason, I incubated...

26 February 2021 9,119 3 View

Will a 10xHis-tag prevent anti-MBP from binding to MBP?

I have a protein with a 10xHis-MBP tag on the N-terminus. I'm fairly certain I am expressing my protein, but I'm not getting a signal on my western blot. Is there a possibility that the 10xHis tag...

25 February 2021 8,404 3 View

Why my agarose gel staining sometimes looks not homogenic but like gradient (see the figure)?

I am worried about this overexposure of the upper part of the gel in the picture. Is it possible that this is the result of too much concentration of ethidium bromide? Why is there such a big...

25 February 2021 8,140 3 View