Overview

  • Product name
  • Description
    Rabbit polyclonal to CX3CR1
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC/IF, Flow Cyt, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide corresponding to Human CX3CR1 aa 2-21 (N terminal).
    Database link: P49238
    (Peptide available as ab8125)

  • Positive control
    • Western Blot: Human spleen tissue lysate, A431, A549, Caco-2, Daudi, HeLa, THP-1 cell lysates. ICC/IF: Human heart cells. IHC-P: Mouse, rat and human heart tissue. Flow cytometry: THP-1 cells. IF: Mouse and rat heart tissue.

Properties

Applications

Our Abpromise guarantee covers the use of ab8021 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
ICC/IF Use a concentration of 10 µg/ml.
Flow Cyt Use a concentration of 0.1 µg/ml.

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

 

WB Use a concentration of 0.5 - 1 µg/ml. Can be blocked with CX3CR1 peptide (ab8125).
IHC-P Use a concentration of 2 µg/ml.

Target

  • Function
    Receptor for the CX3C chemokine fractalkine and mediates both its adhesive and migratory functions. Acts as coreceptor with CD4 for HIV-1 virus envelope protein (in vitro). Isoform 2 and isoform 3 seem to be more potent HIV-1 coreceptors than isoform 1.
  • Tissue specificity
    Expressed in lymphoid and neural tissues.
  • Sequence similarities
    Belongs to the G-protein coupled receptor 1 family.
  • Cellular localization
    Cell membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • Beta chemokine receptor-like 1 antibody
    • C X3 C CKR 1 antibody
    • C-X3-C CKR-1 antibody
    • CCRL1 antibody
    • Chemokine C X3 C motif receptor 1 antibody
    • CMK BRL 1 antibody
    • CMK-BRL-1 antibody
    • CMK-BRL1 antibody
    • CMKBLR1 antibody
    • CMKDR1 antibody
    • CX3C chemokine receptor 1 antibody
    • CX3C CKR1 antibody
    • CX3C1_HUMAN antibody
    • CX3CR1 antibody
    • Fractalkine receptor antibody
    • G-protein coupled receptor 13 antibody
    • GPR13 antibody
    • GPRV28 antibody
    • V28 antibody
    see all

Images

  • All lanes :

    Lane 1 : A431 cell lysates
    Lane 2 : A549 cell lysates
    Lane 3 : Caco-2 cell lysates
    Lane 4 : Daudi cell lysates
    Lane 5 : HeLa cell lysates

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Rabbit IgG antibody (HRP) at 1/10000 dilution

    Developed using the ECL technique.


    10% gel.

    Running conditions: 130v for 2 hours.

    Transfer conditions: wet, 250mA, 2 hrs (Nitrocellulose membrane).

    Blocking condition: 5% non-fat dry milk in TBS, 4C, overnight.

    Primary antibody incubation: Room temperature for 1 hour.

    Secondary antibody incubation: Room temperature for 1 hour. 

    Washing conditions: 15 mL TSBT, 3 x 10 minutes. 

    Exposure: ECL solution

  • Immunohistochemical analysis of paraffin-embedded rat heart tissue labeling CX3CR1 with ab8021 at 2 µg/mL.

  • Immunofluorescence of CX3CR1 in mouse heart tissue with ab8021 at 20 ug/ml. Green: ab8021 Blue: DAPI staining

  • All lanes :

    All lanes : THP-1 cell line

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Rabbit IgG antibody (HRP) at 1/10000 dilution

    Developed using the ECL technique.


    10% gel.

    Running conditions: 130v for 2 hours.

    Transfer conditions: wet, 250mA, 2 hrs (Nitrocellulose membrane).

    Blocking condition: 5% non-fat dry milk in TBS, 4C, overnight.

    Primary antibody incubation: Room temperature for 1 hour.

    Secondary antibody incubation: Room temperature for 1 hour. 

    Washing conditions: 15 mL TSBT, 3 x 10 minutes. 

    Exposure: ECL solution

  • All lanes : Anti-CX3CR1 antibody (ab8021) at 1 µg/ml

    Lane 1 : Human spleen lysate with absence of blocking peptide.
    Lane 2 : Human spleen lysate with presence of blocking peptide.

    Observed band size: 50 kDa
    why is the actual band size different from the predicted?

  • Mouse heart tissue stained for CX3CR1 using ab8021 at 2 µg/ml in immunohistochemical analysis.

  • Flow cytometry analysis of THP-1 cells using CX3CR1 antibody (ab8021) at 0.1 ug/ml. 

    Green: Isotype control

    Red: CX3CR1 antibody (ab8021) 

  • Immunohistochemical staining (paraffin embedded) of human heart tissue using anti-CX3CR1 at 2 µg/ml.
  • Immunofluorescence analysis of rat heart tissue labeling CX3CR1 with ab8021 at 2 µg/mL. DAPI stain in blue. 

  • Immunofluorescence of CX3CR1 in Human Heart cells using ab8021 at 10 ug/ml.

  • Anti-CX3CR1 antibody (ab8021) at 1 µg/ml + Recombinant Human CX3CR1 protein (ab112276) at 0.01 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Exposure time: 4 minutes

References

This product has been referenced in:
  • Tao T  et al. Modulating cAMP responsive element binding protein 1 attenuates functional and behavioural deficits in rat model of neuropathic pain. Eur Rev Med Pharmacol Sci 23:2602-2611 (2019). Read more (PubMed: 30964189) »
  • Wang J  et al. Ischemia-induced Neuronal Cell Death Is Mediated by Chemokine Receptor CX3CR1. Sci Rep 8:556 (2018). Read more (PubMed: 29323156) »
See all 40 Publications for this product

Customer reviews and Q&As

11-19 of 19 Abreviews or Q&A

Answer

Thank you for getting back to me, and I am happy to help resolve this issue. I look forward to hearing from you, and please let me know if you would like any assistance selecting replacements, or if you have any other questions.

Have a great day!

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Answer

Thank you for your enquiry and your interest in our products. All of these 4 antibodies (ab8021, ab7200, ab7201, ab51668) recognize CX3CR1 and are suitable for Western blot application. However, ab7200 has been tested and characterized for detecting rat proteinonly - and not human. The immunogen used to raise this antibody was a recombinant and natural rat CX3CR1 and we do not have any data yet if ab7200 would recognize the human form. It seems that ab8021 is the most popular amongst these antibodies and there are several Abreviews and specific publications listed on the product datasheet. You may wish to take a careful look at it before making the final decision: https://www.abcam.com/cx3cr1-antibody-ab8021.html As you may be aware, RT-PCR data should be treated carefully since, there is no clear correlation between mRNA and protein levels and sometimes it is not easy to predict the protein expression levels from quantitative mRNA data. There are several publications available which delineate the technical boundaries of the approaches for quantitative analysis of protein expression and reveal that simple deduction from mRNA transcript analysis is insufficient. For this reason, I would suggest running a good positive control along with the samples on the same gel; for this purpose tissue lysate from human spleen can be applied. I hope this helps and if I can assist further, please do not hesitate to contact me.

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Question
Answer

Thank you for contacting Abcam. We have a large number of secondary antibodies with fluorescent tags. Theeasiest way to find the one that best fits your needs is from the Abcam home page. The secondary antibodies drop down, located in the yellow banner, has a listing of our DyLight conjugated secondary antibodies. You may then choose the conjugation that is best for your needs. In order to assist your search, I have chosen two products that may work for you. Both are against Rabbit IgG and will work in flow cytometry and IHC. These particular antibodies are conjugated to our DyLight fluorochromes but other fluorochromes are available. The product that I have chosen are ab96891 :https://www.abcam.com/Donkey-polyclonal-Secondary-Antibody-to-Rabbit-IgG-H-L-DyLight-488-ab96891.html and ab96883: https://www.abcam.com/Goat-polyclonal-Secondary-Antibody-to-Rabbit-IgG-H-L-DyLight-488-ab96883.html In regards to your second question. Isotype controls are essential in flow cytometry and recommended in IHC.The appropriate isotype control for this product isab27478. This product has been tested in flow cytometry and IHC-P. I have included the link to a pdf which details our recommended controls for flow cytometry. I hope this information is useful. https://www.abcam.com/ps/pdf/protocols/flow_cytometry_controls.pdf

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Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (THP-1 cells)
Specification
THP-1 cells
Fixative
Acetone
Permeabilization
No

Abcam user community

Verified customer

Submitted Jul 27 2011

Answer

Thank you for contacting us. The CX3CR1 receptor antibody ab8021 will be suitable for flow cytometry analysis of mouse cells. We do not have a directly conjugated antibody against CX3CR1 receptor but we do have conjugation kits. Please see the page at the following link for more details: www.abcam.com/easylink Please do not hesitate to contact us if you need any more advice or information.

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Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Cardiac tissue)
Permeabilization
No
Specification
Cardiac tissue
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative
Acetone

Abcam user community

Verified customer

Submitted Nov 25 2010

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Skin wound)
Specification
Skin wound
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
None

Mr. Josh Meisner

Verified customer

Submitted Aug 19 2010

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (RAW 264.7 Macrophage Cell Line)
Permeabilization
Yes - PBS
Specification
RAW 264.7 Macrophage Cell Line
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 20°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Sep 26 2008

Answer

Thank you for your enquiry. I have attached the protocol below for your reference. Working Citrate Buffer 9mL of 0.1M Citric Acid (10.5g citric acid monohydrate to 500mL DI H2O) 41mL of 0.1M Sodium Citrate (14.7g sodium citrate dehydrate to 500mL DI H2O) 450mL of DI H2O Deparaffinization and Rehydration (Cover staining dishes with a lid in each step) 1. Dip slides in three (3) changes of xylene or a xylene substitute for 3 minutes each. 2. Dip slides in two (2) change of 100% alcohol for 3 minutes each. 3. Dip slides in one (1) change of 95% alcohol for 3 minutes. 4. Dip slides in one (1) change of 70% alcohol for 3 minutes. 5. Rinse slides twice (2x) in DI H2O for 5 minutes. Antigen Retrieval (Microwave Method) 6. Soak slides in 3% H2O2 for 5 minutes. 7. Rinse slides twice (2x) in DI H2O for 5 minutes. 8. Soak the slides in the working citrate buffer and cover with a lid. 9. Microwave until the liquid boils, about 1-5 minutes. 10. Remove from heat and let it stand at room temperature for 20 minutes. 11. Wash three (3) times for 5 minutes in DI H2O 12. Remove the liquid (do not touch the tissue!) and use a PAP pen to circle around the tissue. Blocking 13. Apply enough 5% BSA with a transfer pipette to cover the tissues. 14. Incubate the slides overnight at 4°C in a humid chamber. Primary Antibody 15. Dilute the primary antibody to the recommended concentration in 1% BSA/PBS diluent. 16. Remove the BSA, and incubate with primary antibody solution for 1 hour at room temperature. 17. Wash slides three (3) times 5 minutes each on the shaker. Secondary Antibody and Detection 18. Dilute the biotinylated secondary antibody to 1:200 in 1% BSA diluent. 19. Incubate with secondary antibody solution for 30 minutes at room temperature. 20. Wash slides in PBS three (3) times 5 minutes each on the shaker. 21. Add enough streptavidin HRP to cover the tissues. Incubate for 30 minutes at room temperature 22. Wash three (3) times 5 minutes each in PBS on the shaker. 23. Add enough DAB to cover the tissues. Once the cells start turning brown (inexperienced technicians may wish to observe this under a microscope), wash twice (2x) in PBS for 5 minutes each on the shaker. Optional Counterstain 24. Dip the slide rack with the slides into a staining dish of hematoxylin for 30 seconds. 25. Dip into an acetic bath (200mL DI H2O with one to three drops of acetic acid). Rinse with DI H2O. Dehydration (Cover staining dishes with a lid in each step) 26. Dip slides in 70% and 95% alcohol for 3 minutes each. 27. Dip slides in 2 changes of 100% alcohol for 3 minutes. 28. Dip slides in 3 changes of xylene or xylene substitute for 3 minutes. Cover Slips 29. Drizzle Paramount coverslip solution onto coverslips or slides. 30. Apply coverslip to slide. 31. Let the slides dry overnight.

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11-19 of 19 Abreviews or Q&A

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