Overview

  • Product name

    Anti-CD62P antibody [Psel.KO.2.5]
    See all CD62P primary antibodies
  • Description

    Mouse monoclonal [Psel.KO.2.5] to CD62P
  • Host species

    Mouse
  • Tested applications

    Suitable for: IP, IHC-P, Flow Cytmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Human CD62P transfected 300.19 cells (NP_002996.2).

  • Positive control

    • Human Spleen and Tonsil tissues; Thrombin activated Human platelets.

Properties

Applications

Our Abpromise guarantee covers the use of ab118522 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
IP Use at an assay dependent concentration.
IHC-P Use a concentration of 10 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cyt 1/1 - 1/50.

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

 

Target

  • Function

    Ca(2+)-dependent receptor for myeloid cells that binds to carbohydrates on neutrophils and monocytes. Mediates the interaction of activated endothelial cells or platelets with leukocytes. The ligand recognized is sialyl-Lewis X. Mediates rapid rolling of leukocyte rolling over vascular surfaces during the initial steps in inflammation through interaction with PSGL1.
  • Tissue specificity

    Stored in the alpha-granules of platelets and Weibel-Palade bodies of endothelial cells. Upon cell activation by agonists, P-selectin is transported rapidly to the cell surface.
  • Involvement in disease

    Defects in SELP may be a cause of susceptibility to ischemic stroke (ISCHSTR) [MIM:601367]; also known as cerebrovascular accident or cerebral infarction. A stroke is an acute neurologic event leading to death of neural tissue of the brain and resulting in loss of motor, sensory and/or cognitive function. Ischemic strokes, resulting from vascular occlusion, is considered to be a highly complex disease consisting of a group of heterogeneous disorders with multiple genetic and environmental risk factors.
  • Sequence similarities

    Belongs to the selectin/LECAM family.
    Contains 1 C-type lectin domain.
    Contains 1 EGF-like domain.
    Contains 9 Sushi (CCP/SCR) domains.
  • Cellular localization

    Membrane.
  • Information by UniProt
  • Database links

  • Alternative names

    • Antigen CD62 antibody
    • CD62 antibody
    • CD62 antigen-like family member P antibody
    • CD62P antibody
    • GMP 140 antibody
    • GMP-140 antibody
    • GMP140 antibody
    • GMRP antibody
    • Granule membrane protein 140 antibody
    • Granulocyte membrane protein antibody
    • GRMP antibody
    • LECAM 3 antibody
    • LECAM3 antibody
    • Leukocyte endothelial cell adhesion molecule 3 antibody
    • Leukocyte-endothelial cell adhesion molecule 3 antibody
    • LYAM3_HUMAN antibody
    • P Selectin antibody
    • P-selectin antibody
    • PADGEM antibody
    • Platelet activation dependent granule-external membrane protein antibody
    • Platelet alpha granule membrane protein antibody
    • PSEL antibody
    • Selectin P (granule membrane protein 140kDa, antigen CD62) antibody
    • SELP antibody
    see all

Images

  • Immunohistochemical analysis of CD62P expression in Formalin-Fixed, Paraffin-Embedded Human Spleen using 10 µg/ml ab118522 followed by biotinylated secondary antibody, alkaline phosphatase-streptavidin and chromogen.
  • Immunohistochemical analysis of CD62P expression in Formalin-Fixed, Paraffin-Embedded Human Tonsil using 10 µg/ml ab118522 followed by biotinylated secondary antibody, alkaline phosphatase-streptavidin and chromogen.
  • ab118522 at 10µl/106 cells in 100µl staining CD62P in Thrombin activated Human platelets by Flow cytometry.

References

This product has been referenced in:

  • Preidl RHM  et al. Long-term endothelial dysfunction in irradiated vessels: an immunohistochemical analysis. Strahlenther Onkol 195:52-61 (2019). Read more (PubMed: 30324290) »
See 1 Publication for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Question

Our customer has answered your questions below in blue. Also they tried the antigen retrieval steps and did not get any improvement.

Could you advise on how to proceed?

Best regards,


I have read the details customer have kindly provided. However I have few other questions;

- Does both GR62917-2 AND GR62917-3 lot numbers were negative?Yes, both lot numbers were negative, no staining at all.
- What is the species of secondary antibody?As secondary antibody we use Envision reagent from DAKO.The EnVision™ reagent of this kit is a peroxidase-conjugated polymer backbone, which, in addition, also carries secondary antibody molecules directed against rabbit and mouse immunoglobulins. The combination of several peroxidase molecules and several secondary antibody molecules on the same polymer provides a simple, yet sensitive, visualization system.
- Does negative mean no staining at all or high background? We see no staining at all.
- Have you tried optimizing of antigen retrieval step; the optimization is necessary when antibodies gives no staining or high background? No, weonly did antigen retrieval for 15 minutes in Citrate buffer inmicrowave.
- Have you tried incubating the sections overnight? No, we did not.
- For how long the tissues were fixed in formalin? The time of fixation in formalin was about 4 hours.

Recommendations for further optimizations

-Try antigen retrieval for 5,10,15,20 minutes by incubating the section in microwave (700 watt) with citrate buffer pH6.0. The time range will help getting the optimized retrieval time.
- Check the pH of buffer, antibodies don't work if the buffer pH is high or low.
- Incubated the primary overnight at 4C.
- The tissue should not be fixed for more than 24 hours.
- Add 0.2% Tween in the PBS for antibody dilution, blocking solution and washing solution.
- Block the sections with 10% normal serum or 5% BSA for 1-2 hours.

I hope these troubleshooting suggestions will help to get good staining of target protein. Please do not hesitate to contact me if the results do not improve.




.

Read More
Answer

Thank you for your response.
Could you please confirm the order details please.

Date of purchase,

- Abcam Order Number or

- Your PON

Question: Has the customer used the same detection system (peroxidase-conjugated polymer from EnVision™) successfully with a different primary antibody?

I look forward to hearing from you soon.

Read More

Question

We have forwarded your questions to our client and they have answered as follows:


I have read the details customer have kindly provided. However I have few other questions;

- Does both GR62917-2 AND GR62917-3 lot numbers were negative?Yes, both lot numbers were negative, no staining at all.
- What is the species of secondary antibody?As secondary antibody we use Envision reagent from DAKO.The EnVision™ reagent of this kit is a peroxidase-conjugated polymer backbone, which, in addition, also carries secondary antibody molecules directed against rabbit and mouse immunoglobulins. The combination of several peroxidase molecules and several secondary antibody molecules on the same polymer provides a simple, yet sensitive, visualization system.
- Does negative mean no staining at all or high background? We see no staining at all.
- Have you tried optimizing of antigen retrieval step; the optimization is necessary when antibodies gives no staining or high background? No, weonly did antigen retrieval for 15 minutes in Citrate buffer inmicrowave.
- Have you tried incubating the sections overnight? No, we did not.
- For how long the tissues were fixed in formalin? The time of fixation in formalin was about 4 hours.

Recommendations for further optimizations

-Try antigen retrieval for 5,10,15,20 minutes by incubating the section in microwave (700 watt) with citrate buffer pH6.0. The time range will help getting the optimized retrieval time.
- Check the pH of buffer, antibodies don't work if the buffer pH is high or low.
- Incubated the primary overnight at 4C.
- The tissue should not be fixed for more than 24 hours.
- Add 0.2% Tween in the PBS for antibody dilution, blocking solution and washing solution.
- Block the sections with 10% normal serum or 5% BSA for 1-2 hours.

I hope these troubleshooting suggestions will help to get good staining of target protein. Please do not hesitate to contact me if the results do not improve.

They are currently looking into optimizing their antigen retrieval steps and the other tips you have kindly provided.

We look forward to your response.

Best regards,

Read More
Answer

Thank you for your email. Please let me know how the staining go after following the troubleshooting suggestions.

Read More

Answer

Thank you for your email.

I have read the details customer have kindly provided. However I have few other questions;

- Does both GR62917-2 AND GR62917-3 lot numbers were negative?
- What is the species of secondary antibody?
- Does negative mean no staining at all or high background?
- Have you tried optimizing of antigen retrieval step; the optimization is necessary when antibodies gives no staining or high background?
- Have you tried incubating the sections overnight?
- For how long the tissues were fixed in formalin?

Recommendations for further optimizations

-Try antigen retrieval for 5,10,15,20 minutes by incubating the section in microwave (700 watt) with citrate buffer pH6.0. The time range will help getting the optimized retrieval time.
- Check the pH of buffer, antibodies don't work if the buffer pH is high or low.
- Incubated the primary overnight at 4C.
- The tissue should not be fixed for more than 24 hours.
- Add 0.2% Tween in the PBS for antibody dilution, blocking solution and washing solution.
- Block the sections with 10% normal serum or 5% BSA for 1-2 hours.

I hope these troubleshooting suggestions will help to get good staining of target protein. Please do not hesitate to contact me if the results do not improve.

Read More

Answer

Thank you for contacting us.

I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

I am attaching our questionnaire so that we can gather further information regarding the samples tested and the protocol used. Once we receive the completed questionnaire, we will look at the protocol and see if there are any suggestions we can make that may improve the results. This information will also allow us to investigate this case internally and initiate additional testing where necessary. If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody.

I look forward to receiving your reply.

Read More

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