Overview

  • Product name
    Anti-Ly6g antibody [RB6-8C5]
    See all Ly6g primary antibodies
  • Description
    Rat monoclonal [RB6-8C5] to Ly6g
  • Host species
    Rat
  • Tested applications
    Suitable for: IHC-Fr, IP, , Functional Studies, IHC-P, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse
  • Immunogen

    Tissue/ cell preparation of Normal murine bone marrow (Mouse)

  • General notes

    This product was changed from ascites to tissue culture supernatant on 3rd September 2018. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

Properties

Applications

Our Abpromise guarantee covers the use of ab25377 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
IHC-Fr Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
Depletion Use at an assay dependent concentration.

In vivo depletion of Ly-6G+ cells.

Functional Studies Use at an assay dependent concentration.

In vitro complement-dependent cytotoxicity assays.

IHC-P Use at an assay dependent concentration. PubMed: 25126943
Flow Cyt 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

  • ab25377 staining Ly6g in Mouse placenta tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 5% BSA for 30 minutes at 25°C; antigen retrieval was by heat mediation. Samples were incubated with primary antibody (1/100 in 1% BSA in PBS) for 12 hours at 4°C. ab6734,  a HRP-conjugated Rabbit anti-rat IgG polyclonal (1/200) was used as the secondary antibody.

     

    See Abreview

  • Immunohistochemistry (Frozen sections) analysis of Mouse frozen spleen tissue labeling Ly6g with ab25377. Right: Rat Anti-Mouse Ly-6G-UNLB followed by Mouse Anti-Rat IgG2b-FITC. Left: Isotype Control, Rat IgG2b-UNLB followed by Mouse Anti-Rat IgG2b-FITC. Blue - DAPI.

  • ab25377 staining Ly6C in Mouse heart tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 0.5% BSA; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/100) for 1 hour at room temperature. A Peroxidase-conjugated anti-rat Ig polyclonal was used as the secondary antibody.

  • ab25377 staining Ly6g in Mouse bone tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with PBS, BSA 1% + NGS5% for 30 minutes at 20°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/200 in PBS + BSA 1%) for 9 hours at 4°C. A Biotin-conjugated Goat anti-rat polyclonal (1/500) was used as the secondary antibody.

    See Abreview

References

This product has been referenced in:
  • Thapa D  et al. The Cannabinoids ?8THC, CBD, and HU-308 Act via Distinct Receptors to Reduce Corneal Pain and Inflammation. Cannabis Cannabinoid Res 3:11-20 (2018). Read more (PubMed: 29450258) »
  • Li Z  et al. FOXO3-dependent apoptosis limits alcohol-induced liver inflammation by promoting infiltrating macrophage differentiation. Cell Death Discov 4:16 (2018). Read more (PubMed: 29531813) »
See all 45 Publications for this product

Customer reviews and Q&As

1-10 of 10 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
PBS BSA 1% NGS 5% as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 20°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate
Sample
Mouse Tissue sections (bone)
Specification
bone
Permeabilization
No
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Mar 24 2014

Answer

We state to perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol as done in PubMed: 21713436. This publication used a citrate buffer pH6 so we would recommend you try this buffer also.

Read More
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Antigen retrieval step
Heat mediated
Sample
Mouse Tissue sections (placenta)
Specification
placenta
Permeabilization
No
Fixative
Formaldehyde

Dr. Varkha Agrawal

Verified customer

Submitted Nov 11 2013

Answer

Thank you for your inquiry.

I am happy to confirm that ab25377 is tested and guaranteed for mouse samples and IHC-P. WE do not know of a special protocol for distal colon from mouse and have not tested this specifically. Maybe the literature (https://www.abcam.com/Ly6g-antibody-RB6-8C5-ab25377-references.html) can provide more information.

For general detailed IHC protocol I can recommend our Protocol pages:

https://www.abcam.com/index.html?pageconfig=popular_protocols

https://www.abcam.com/index.html?pageconfig=resource&rid=11384

I send you a quote for 100ug (1 vial) of ab25377 in a separate email.

I hoe this information is helpful and wish you good luck with your research.

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Answer

Thank you for taking the time to contact us. I am sorry to hear you have had difficulty obtaining satisfactory results with the Anti-Ly6g antibody [RB6-8C5] (ab25377).

I would like to reassure you that this antibody is tested and covered by our 6 month guarantee for mouse samples and IHC-P. In the event that a product is not functioning in the tested applications and species cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund.

I would like to investigate this particular case further for you, and also obtain more information for our quality monitoring records. I would appreciate if you could also provide an image which would help us to assess the results.

Reviewing the provided information, I would like to offer some suggestions. I would also appreciate if you can confirm some further details:

1. Could you please confirm the order number and date of purchase. Was it order 1103213 on the 07/07/2012?

2. Please provide information about how the antibody was stored?

3. Could you confirm what level of expression you would expect in inflamed mouse skin tissue? I was searching for literature but regrettably could not find any reference to confirm the expression level of Ly6g in mouse inflamed skin tissue. Do you have any information regarding the expression level of Ly6g in this sample type? I can suggest it would be beneficial to consider a positive control if not already performed? References we are aware of for this product are cited on our product web page https://www.abcam.com/Ly6g-antibody-RB6-8C5-ab25377-references.html. I hope they might be helpful to find a positive control which is suitable for the experiment performed.

4. Could you please confirm the isotype which is recognised by the secondary antibody that was used. Does the secondary antibody detect rat IgG2b?


Thank you for your time and cooperation. I look forward to hearing from you with the requested details. I hope we can resolve this case for you as soon as possible.

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Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number xxx with ab45977. The item is set for shipping on monday.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

Read More

Answer

I have performed a BLAST search with the Ly6g sequence, and it looks like the sequence of Ly6g and Ly6c are 60 to 64% identical, it is rather unlikely that this antibody will react withLy6c:We recommend that alignment should be over 85% to predict that an antibody will detect in a different species or different protein isoforms.

However, we would be unable to guarantee this without further testing.

Read More

Answer

Thank you for contacting us.

Yes, has been used successfully on mouse, and as well in IHC-P. therefore, you are covered by our Abpromise.

We haven't tested IHC-P ourselves, but the following reference used the antibody in IHC-P under the following conditions:

From Król M et al. Density of Gr1-positive myeloid precursor cells, p-STAT3 expression and gene expression pattern in canine mammary cancer metastasis. Vet Res Commun 35:409-23 (2011)."Five um sections from paraffin blocks containing tumor tissue were baked in 37°C overnight. After dewaxing in xylene and rehydration in ethanol, for antigen retrieval, the slides were placed in 0.02 M citrate buffer, pH 6.0 and boiled in the decloaking chamber."

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

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Answer

Thank you for contacting us.

For ab25377, I would recommend a starting WB concentration of 1ug/mL. The final concentration will need to be optimized to your specific protocol and samples.

This clone has been used in a number of references, which are listed in the "Specific references" section of our datasheet. Unfortunately, I do not have access to all of the journals so I am not sure which of these antibodies use RB6-8C5 in WB.

I hope this helps, please let me know if you have any additional questions.

Read More

Answer

Thank you for your call yesterday and for your patience while I have looked into your enquiry.

I have found a couple of potential markers: beta glucan receptor and neutrophil elastase. Beta glucan receptor is expressed by cells of the monocyte/macrophage and neutrophil lineages, but also at lower levels by dendritic cells and a subpopulation of T cells, so you may be able to optimize the dilution factor so that dendritic cells are not stained.

I didn't find conclusive evidence that neutrophil elastase is not expressed by dendritic cells, but I did find several articles about the affect of neutrophil elastase on dendritic cells, so I'm assuming this molecule is secreted by other cells.

We do have a beta glucan receptor antibody that will react with mouse, ab53427, however it will not work in formalin fixed paraffin embedded tissue, only in frozen tissue-

https://www.abcam.com/beta-Glucan-Receptor-antibody-2A11-ab53427.html

For neutrophil elastase, I would recommend ab68672, which will work in IHC-P or IHC-Fr-

https://www.abcam.com/Neutrophil-Elastase-antibody-ab68672.html

As I couldn't find conclusive evidence about these markers, I encourage you to look through the literature to ensure that these markers are suitable for your study.

I hope this information will be useful, but if you have any further questions or if there is anything else that we can do for you, please let me know and I'll be happy to help.

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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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