Marcia Chavez

11 Questions 14 Answers 0 Followers

Questions related from Marcia Chavez

How difficult is it to detect 3 targets with an ELISA?

I'm new to ELISA assays and have heard that multiplexing can be difficult. In your experience, what are some of the challenges that come with multiplexing? Is detecting up to 3 targets doable?

22 June 2023 1,349 3 View

What could be causing these squiggles in immunofluorescence?

I have these squiggles that are appearing in my GAD67 IF stain and I'm not sure why. I've used this secondary antibody (Cy5 goat anti-mouse) with another primary antibody and never had this...

27 June 2022 2,052 9 View

Do incubation times differ for monoclonal vs polyclonal antibodies in IF?

I'm getting a lot of non-specific staining with a polyclonal ERa antibody I'm using. There's so much of it that I'm beginning to think that my incubation period (48 hrs at 4C) is too much....

07 March 2022 7,251 3 View

Can background be removed with ImageJ's rolling ball radius tool?

Can the rolling ball radius tool be used alone to reduce background in fluorescent images? Why or why not? If not, what are some additional methods to further remove background after rolling ball...

24 February 2022 3,850 3 View

Alternatives to Minocycline?

I'm looking for alternatives to Minocycline that would be relatively safe to administer during pregnancy in a rodent model. So far, I've only found one paper that gives this drug prenatally and no...

05 November 2021 455 1 View

Is there a way to section mutliple rat brains at the same time?

Aside from embedding brains in an OCT block, are there other ways to section multiple brains at once?

19 June 2021 4,009 4 View

Do tissue samples need to be reblocked between primary antibody applications?

In other IHC double-label protocols, I see that people reblock the tissue between primary antibody applications. Is this really necessary? Reblocking makes sense if the secondaries of the two...

22 April 2021 4,920 1 View

What are these "dots" in the background of my IF stain?

I recently tried an ERa immunofluorescent stain using strep488 as the secondary. There are clearly ERa positive cells, but I'm not sure what these "dots"/noise are in the background of my GFP channel.

22 March 2021 7,053 10 View

Does anybody have an immunofluorescent protocol for ERα using whole brain sections?

I've been troubleshooting my IF ERα protocol and I'm curious to see what reagents other people use in this stain. Thanks.

01 March 2021 4,537 1 View

Can I use carbo-free blocking solution in place of normal goat serum?

I'm looking to try this in place of normal goat serum for a wisteria floribunda lectin stain. In my IF protocol, 10% of the final volume of my blocking solution is goat serum. If I were to go the...

08 February 2021 7,665 6 View

Can problems with blocking cause high background and weak antibody signal during immunofluorescence staining?

I'm working with 40um fresh frozen (non-perfused) sections of rat brain tissue and I'm staining for wisteria lection (immunofluorescence). After staining, my tissue has high background and a weak...

11 November 2020 3,751 7 View