Overview

  • Product name

    ER Staining Kit - Red Fluorescence - Cytopainter
    See all Endoplasmic reticulum kits
  • Sample type

    Adherent cells, Suspension cells
  • Assay type

    Cell-based
  • Species reactivity

    Reacts with: Mammals, Other species
  • Product overview

    ER Staining Kit - Red Fluorescence | Cytopainter (ab139482) contains an endoplasmic reticulum-selective red dye suitable for live cell, or detergent-permeabilized aldehyde-fixed cell staining. Micromolar concentrations of the red dye are sufficient for staining mammalian cells, as validated with human cervical carcinoma cell line HeLa, human T-lymphocyte cell line Jurkat and human bone osteosarcoma epithelial cell line, U2OS.


    One important application of the ER staining kit is in fluorescence co-localization imaging with green fluorescent protein (GFP)-tagged proteins, a powerful approach for determining the targeting of molecules to intracellular compartments and for screening of their associations and interactions.


    The ER staining kit is specifically designed for use with GFP-expressing cell lines, as well as cells expressing blue, cyan or yellow fluorescent proteins (BFPs, CFPs, YFPs). Additionally, the kit is suitable for use with live or post-fixed cells in conjunction with probes, such as labeled antibodies, or other fluorescent conjugates displaying similar spectral properties as fluorescein, or coumarin.


    A nuclear counterstain is also included in the kit.


    Review other dyes and kits for ER staining, or the live cell staining fluorescent dyes guide

  • Notes

    This kit can be readily used in combination with other common UV and visible light excitable organic fluorescent dyes and various fluorescent proteins in multi-color imaging and detection applications. The dye emits in the Texas Red region of the visible light spectrum, and is resistant to photo-bleaching, concentration quenching and photoconversion.

    Previously called CytoPainter ER Staining Kit -Red Fluorescence.

  • Platform

    Fluorescence microscope

Properties

Images

  • Live HeLa cells stained with Red Detection Reagent (A), Hoechst dye (B) and resulting composite image (C).
  • Fluorescence excitation and emission spectra for Red dye (panel A) and absorbance and fluorescent emission spectra for Hoechst 33342 dye (panel B). All spectra were determined in 1X Assay Buffer.

Protocols

References

This product has been referenced in:

  • Liu T  et al. Glycosylation controls sodium-calcium exchanger 3 sub-cellular localization during cell cycle. Eur J Cell Biol 97:190-203 (2018). Read more (PubMed: 29526322) »
See 1 Publication for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Answer

Staining at low concentrations does not appear to be toxic to cells. I know that you already tried to reduce the concentration, maybe you can try to reduce it more.

Customer feedback: I've tested the ER staining on Friday and the 0.2X concentration can stain HeLa cells well without causing too much toxicity. It is good new but we spent quite some effort to obtain the same results in HeLa as you advertise on the website. In contrast, the staining using Mitochondria and lysosome Cytopainter were just as you described. We've used many of your products and this ER stain protocol was the least satisfying one so far. Hope the information could be modified and cost less optimisation time in future.

Read More

Combining ER dye and vimentin antibody staining

Good Excellent 5/5 (Ease of Use)
Abreviews
HUVEC (30000) were seeded 8-well ibiTreat ibidi culture slides, and grown until nearly confluent for 24-48h.
Cells were washed with PBS, fixated with 1% PFA for 20' RT and subsequently permeabilized with PBS/0.1%Triton-X100 for 10’RT.
Cells were incubated with primary Ab (mouse anti-vimentin, clone V9, Dako, 1:200), secondary goat-anti-mouse biotin (Dako, 1:200) and streptavidin-Alexa488 (ThermoFisher 1:500), all diluted in PBS/0.5%BSA. All incubations were for 45-60' RT in the dark, followed by 3 washes with PBS.
Cytopainter ER staining dye was diluted 1:1000 in 1x assay buffer, added to the cells, and incubated for 30' at 37C. If applicable, Hoechst was also diluted 1:1000 in the same buffer.
Cells were washed 2x in PBS before images were taken on a Leica DMIL inverted microscope equipped with a Leica DFC345FX camera, and processed in Leica Application Suite software.

Figure:
HUVEC stained with Cytopainter ER staining dye (red) Hoechst (blue) and anti-vimentin Ab (green).

Judy Van Beijnum

Verified customer

Submitted Oct 26 2017

Chicken Fibroblasts

Good Excellent 5/5 (Ease of Use)
Abreviews
Chicken Embryonic Fibroblasts (CEF) stained according to Abcam's recommended protocol.

Dr. Marcelo Pelajo Machado

Verified customer

Submitted May 09 2017

Economic and very easy to use

Good Excellent 5/5 (Ease of Use)
Abreviews
1. Mouse astrocyte primary culture was fixed in 4%PFA for 5 minutes.
2. Block and permeabilized o/n, 4C with 1%BSA+0.1% Triton-X.
3. In 1X buffer: 1:1000 Texas red (ER) and 1:1000 Hoestch, room temperature for 40 minutes,
4. Rinse with 1X buffer twice and mount.
Imaged in a Leica SP8, 40X magnification, 1024x1024.

Ms. Maria Velasco

Verified customer

Submitted Mar 17 2017

ab139482 for mouse tissue

Good Excellent 5/5 (Ease of Use)
Abreviews
This kit is good!

Abcam user community

Verified customer

Submitted Jun 18 2015

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up