Overview

  • Product name

    Anti-CD14 antibody [2D-15C, FMC-32]
    See all CD14 primary antibodies
  • Description

    Mouse monoclonal [2D-15C, FMC-32] to CD14
  • Host species

    Mouse
  • Specificity

    The clone number has been updated from (2Q1233) to (2D-15C, FMC-32) both clone numbers name the same antibody clone.
  • Tested applications

    Suitable for: Flow Cyt, ICC/IF, IHC-Frmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Human peripheral blood mononuclear cells.

  • Positive control

    • Peripheral blood monocytes.
  • General notes

    Though we have received a positive Abreview for this antibody in IHC-P, we have also received many negative reports in IHC-P and so we can no longer guarantee this antibody in this application. We appreciate any further feedback from users in this regard.

    ab63319 reacts with peripheral blood monocytes and weakly with granulocytes. It is negative to platelets, erythrocytes and lymphocytes. It is sometimes positive for chronic myeloid leukemia (CML) and acute myeloid leukemia (AML). It is negative to B-cell chronic lymphocytic leukemia (B-CLL), T-cell chronic lymphocytic leukemia (T-CLL) and common acute lymphoblastic leukemia (CALL).

     

Properties

Applications

Our Abpromise guarantee covers the use of ab63319 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 1/25.

Dilute with isotonic buffer. Use 50ul per 1x106 peripheral blood mononuclear cells (PBMC) in 100ul buffer.

 

 

 

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 

ICC/IF 1/25.
IHC-Fr 1/25. We recommend acetone fixation.

Target

  • Function

    Cooperates with MD-2 and TLR4 to mediate the innate immune response to bacterial lipopolysaccharide (LPS). Acts via MyD88, TIRAP and TRAF6, leading to NF-kappa-B activation, cytokine secretion and the inflammatory response. Up-regulates cell surface molecules, including adhesion molecules.
  • Tissue specificity

    Expressed strongly on the surface of monocytes and weakly on the surface of granulocytes; also expressed by most tissue macrophages.
  • Sequence similarities

    Contains 11 LRR (leucine-rich) repeats.
  • Post-translational
    modifications

    N- and O- glycosylated. O-glycosylated with a core 1 or possibly core 8 glycan.
  • Cellular localization

    Cell membrane.
  • Information by UniProt
  • Database links

  • Alternative names

    • CD 14 antibody
    • CD_antigen=CD14 antibody
    • CD14 antibody
    • CD14 antigen antibody
    • CD14 molecule antibody
    • CD14_HUMAN antibody
    • LPS-R antibody
    • Mo2 antibody
    • Monocyte differentiation antigen CD14 antibody
    • Monocyte differentiation antigen CD14 urinary form antibody
    • Monocyte differentiation antigen CD14, membrane-bound form antibody
    • Myeloid cell specific leucine rich glycoprotein antibody
    • Myeloid cell-specific leucine-rich glycoprotein antibody
    see all

References

This product has been referenced in:

  • Erikson E  et al. In vivo expression profile of the antiviral restriction factor and tumor-targeting antigen CD317/BST-2/HM1.24/tetherin in humans. Proc Natl Acad Sci U S A 108:13688-93 (2011). Flow Cyt ; Human . Read more (PubMed: 21808013) »
  • Alm JJ  et al. Circulating plastic adherent mesenchymal stem cells in aged hip fracture patients. J Orthop Res 28:1634-42 (2010). Read more (PubMed: 20540091) »
See all 2 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Application
Immunohistochemistry (Frozen sections)
Sample
Sheep Tissue sections (Cardiac tissue)
Permeabilization
No
Specification
Cardiac tissue
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
Acetone

Abcam user community

Verified customer

Submitted Jun 08 2017

Application
Flow Cytometry
Sample
Human Cell (Peripheral macrophages)
Specification
Peripheral macrophages
Preparation
Cell harvesting/tissue preparation method: Ficoll gradient separation/adhesion
Sample buffer: PBS
Permeabilization
No
Gating Strategy
FL1

Abcam user community

Verified customer

Submitted Mar 18 2013

Answer

According to our records, ab39256, ab63319, ab77258, ab86673, ab103674 was proving difficult to use in [APPL] and we were in contact in order to help resolve the issue.

Looking at our correspondence, it appears that we are awaiting more details in order to help us better understand the difficulties experienced. If the requested information has already been sent, it appears that it did not reach our Scientific Support team and we apologize for this inconvenience. In this case we would like to ask for the information again so that we can reach a resolution.

If the issue has already been settled, please let us know so that we can be assured that the problem has been solved to your satisfaction and update our records.

We wish you the best of luck with your research and look forward to a reply.

Read More

Answer

Thank you for your email.

We will look forward to receive the images. On the optimization note we strongly recommend to include the blocking step in protocol; please try 5%BSA or 5-10% normal serum for 1-2 hours at room temperature with species same as secondary antibody. I am sure this step would improve the staining with ab103674 and ab39256.

I may be that the tissue sections you are using does not express the targets GCET1, ERG, CD14 or express at very low level so could you provide the details tissue type and species of tissue section tested. We are interested in knowing e.g. human brain tissue sections or house heart tissue sections etc.

I would suggest trying range of incubation with microwave e.g. for 5, 10, 15 and 20 minutes. This range would help to get the optimized incubation at which the antigen is maximum exposed.

I hope these suggestions would be helpful. Should you have any further question please do not hesitate to contact us.

Read More

Answer

Thank you for your email. I am sorry to hear that you have been experiencing problems with this antibody.

I have read the details you have kindly provided and have following further questions for better understanding of the problem;

- I am very surprised that 5 antibodies failed in row? Could you please specify which other antibody worked?
- What is the type and species of tissue used with each antibody? Could you supply staining images?
- Are the tissues paraffin fixed as mentioned in the details or paraformaldehyde fixed and paraffin embedded sections?
- What was the pH of Tris and Citrate buffer?
- Did tyou permeabilize the tissue sections by incubating them with Triton X-100.
- Metanol/ H2O2 blocking used to block the endogenous peroxidases, which gives false staining when HRP conjugated secondary is used. Have you done the blocking with Normal serum or BSA.
- What dilution of primaries were used in each experiment?
- What is the catalogue number of DAKO envision secondary antibody use? Did you use same kit for all the antibodies? What were the other secondary antibodies used?
- Have you used this detection system successfully before?
- Did you tired optimizing the antigen retrieval method?

Please note more information you will provide-resolution will be achieved quickly.

Thank you very much for your cooperation. I will look forward to hearing from you soon.

Read More

Question
Answer

Thank you for your email.

I am sorry this product did not perform in IHC-P as originally 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 ab24590.

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
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (tonsil)
Specification
tonsil
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: EDTA, pH8, 100C, 20 minutes
Permeabilization
No

Abcam user community

Verified customer

Submitted Dec 29 2009

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