Overview

  • Product name
    Anti-Ly6g antibody [RB6-8C5] (FITC)
    See all Ly6g primary antibodies
  • Description
    Rat monoclonal [RB6-8C5] to Ly6g (FITC)
  • Host species
    Rat
  • Conjugation
    FITC. Ex: 493nm, Em: 528nm
  • Specificity
    ab25024 recognises mouse Mouse Ly-6G.
  • Tested applications
    Suitable for: Flow Cyt, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Monkey
  • Immunogen

    The details of the immunogen for this antibody are not available.

  • Positive control
    • mouse spleen cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab25024 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Flow Cyt Use at an assay dependent concentration.

ab136125 - Rat monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

 

IHC-Fr Use at an assay dependent concentration.

Target

  • Relevance
    Ly6G is a GPI-anchored protein, that is also known as the myeloid differentiation antigen Gr1. The antigen is transiently expressed on monocytes in the bone marrow. The level of antigen expression in the bone marrow directly correlates with granulocyte differentiation and maturation. Ly6G is expressed predominantly on neutrophils, also in a subset of eosinophils, differentiating pre-monocytes, and plasmacytoid dendritic cells.
  • Cellular localization
    Attached to the membrane by a GPI-anchor.
  • Database links
  • Alternative names
    • Gr-1 antibody
    • Gr1 antibody
    • Ly-6G.1 antibody
    • Ly6G antibody
    • Lymphocyte antigen 6 complex locus G antibody
    • Lymphocyte antigen 6G antibody
    see all

Images

  • Flow Cytometry analysis of C57Bl/6 bone marrow cells labeling Ly6g with ab25024 0.5µg (right panel), Rat IgG2b (FITC) isotype control at 0.5µg (left panel).

  • BALB/c bone marrow cells were stained with either rat IgG2b-FITC (as an isotype control) or rat anti-mouse Ly-6G-FITC (ab25024) at a concentration of 1ug/106 cells.  Large cells were then gated and analyzed on a flow cytometer.

  • Flow Cytometry analysis of C57Bl/6 bone marrow cells labeling Ly6g with ab25024 at 0.5µg.

  • ab25024 at a 1/100 dilution detecting Ly6g in LPS activated monocytes by Flow Cytometry.
    Gating strategy was for alive cells.

    See Abreview

  • ab25024 at a 1/100 dilution detecting Ly6g (Gr1) in murine blood cells by Flow Cytometry.
    Gating strategy was for monocytes.

    See Abreview

  • ab25024 staining Ly6g in Cynomolgus Monkey whole blood by Flow Cytometry. Red blood cells were lysed in PBS + 1% BSA and 0.01% sodium azide and fixed in paraformaldehyde. The sample was incubated with the primary antibody (1/10 in PBS + 1% BSA and 0.01% sodium azide) for 1 hour at 4°C.
    Gating Strategy: Live Monocytes.

    See Abreview

  • ab25024 detecting Ly6g in rat spleenocytes by Flow Cytometry. Spleen tissue was homogenized and red blood cells were lysed in PBS + 1% BSA and 0.01% sodium azide. The sample was incubated with the primary antibody (1/100 in PBS + 1% BSA and 0.01% sodium azide) for 1 hour at 4°C.
    Gating Strategy: Live macrophages.

    See Abreview

References

This product has been referenced in:
  • Lautz T  et al. Midkine Controls Arteriogenesis by Regulating the Bioavailability of Vascular Endothelial Growth Factor A and the Expression of Nitric Oxide Synthase 1 and 3. EBioMedicine 27:237-246 (2018). Read more (PubMed: 29233575) »
  • Wen G  et al. Genetic and Pharmacologic Inhibition of the Neutrophil Elastase Inhibits Experimental Atherosclerosis. J Am Heart Assoc 7:N/A (2018). Read more (PubMed: 29437605) »
See all 5 Publications for this product

Customer reviews and Q&As

1-4 of 4 Q&A

Answer

Having reviewed this case, I would like to offer some suggestions to help optimize the results from ab25024. I would also appreciate if you can confirm some further details: 1) I see in your details that you are using 0.75ul (0.375ug) of antibody for your stain. We recommend use of 1ug (2ul) of antibody per 1x10^6 cells, have you attempted to use a higher concentration (2ul) per sample? 2) Longer incubation may be necessary for optimal binding of the antibody in your samples. We would recommend trying a 45 minute of 1 hour incubation to give this item more time to bind the expressed Ly6g. 3) Wash the cells 3 X by centrifugation in FACS buffer at 400 g for 5 minutes and resuspend them in 500 µl to 1 ml of ice cold FACS buffer and store cold in dark until analysis. 4) The light fixation may also be interfering with this product, have you have the chance to run this without using 1% pfa?

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Answer

Thank you for submitting an Abreview for ab25024. As you may have noticed, your review has now been published on our website.
Since you obtained poor results using the antibody in an untested species (dog), we would like to follow up on this to see if we can possibly improve the results you are seeing with this antibody.
Based on the information in your review, I feel that altering the following step in your protocol may help improve your results:
- fix with PFA after incubation with the primary antibody - if you haven't done that already.
- incubate longer with the primary antibody (e.g. 2-3 h at RT or overnight at 4 degree)
It could also just be the case that the antibody does not work for dog samples due to potential low homology.
If you decide to follow these suggestions, please let me know if they are helpful.
I wish you good luck with your research.

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Question
Answer

Thank you for your patience.


I have now had the Rat IgG1 (FITC) isotype control added to our catalog, ab128477.


I hope this information is helpful. Please do not hesitate to contact us if you have any additional questions.

Read More

Question
Answer

Thank you for contacting Abcam regarding ab25024.


I am working to have the appropriate isotype control added to our catalog for you to purchase. I will provide the ordering details as soon as they are available.


I hope this information is helpful. Please do not hesitate to contact me if you have any additional questions.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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