Overview

  • Product name

    Human PF4 ELISA Kit (CXCL4)
    See all PF4 kits
  • Detection method

    Colorimetric
  • Sample type

    Cell culture supernatant, Serum, Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    < 20 pg/ml
  • Range

    20.58 pg/ml - 15000 pg/ml
  • Recovery

    100 %

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 108.2 101% - 116%
    Serum 122.4 110% - 130%
    Plasma 119.6 111% - 127%

  • Assay duration

    Multiple steps standard assay
  • Species reactivity

    Reacts with: Human
  • Product overview

    Abcam’s PF4 (CXCL4) Human ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of Human PF4 in serum, plasma, and cell culture supernatants.


    This assay employs an antibody specific for Human PF4 coated on a 96-well plate. Standards and samples are pipetted into the wells and PF4 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-Human PF4 antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of PF4 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.


    Get results in 90 minutes with Human PF4 ELISA Kit (CXCL4) (ab189573) from our SimpleStep ELISA® range.

  • Notes

    Optimization may be required with urine samples.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Microplate

Properties

Associated products

Applications

Our Abpromise guarantee covers the use of ab100628 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
Sandwich ELISA Use at an assay dependent concentration.

Images

  • Human PF4 measured in biological fluids showing quantity (pg) per mL of tested sample. Samples were diluted 200-800 fold.

  • Representative Standard Curve using ab100628.

Protocols

References

This product has been referenced in:

  • Yang J  et al. Peptidome Analysis Reveals Novel Serum Biomarkers for Children with Autism Spectrum Disorder in China. Proteomics Clin Appl 12:e1700164 (2018). Read more (PubMed: 29754444) »
See 1 Publication for this product

Customer reviews and Q&As

1-9 of 9 Abreviews or Q&A

Question
Answer

One of the two vials of the PF4 (CXCL4) standard should be reconstituted in 400 uL Assay diluent C (for serum/plasma samples) or 1X Assay Diluent B (for cell culture medium and urine). This will yield a 140 ng/ml stock standard solution. The second vial is an extra vial should you need to run the assay again, so you can prepare a fresh standard dilution.

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Question
Answer

The 2nd PF4 Standard vial is just an extra vial. It is there to minimize freeze-thaw cycles in case you want to perform multiple experiments with the kit on different days.

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Answer

Thank you for your inquiry regarding ab100628.

Unfortunately, we do not know whether this PF4 Human ELISA kit will also recognize the bound PF4-heparin complex form. All we know is that the antibody pair is binding on the PF4 protein somewhere between aa 32-101 (which is the full length protein minus the signal peptide). Therefore, whether the kit will also detect the bound PF4-heparin complex form would need to be determined by the customer.

If you need any further assistance in the future, please do not hesitate to contact me.

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Answer

Thank you for contacting us this afternoon and for your interest in Abcam.

Following on from our phone call I have discussed with my colleague xxxxx in marketing whether we have any start-up offers available. He informed me that this is something that is not routinely done but he would be happy to speak with you in further detail as to what you are looking for. I have CC'd him in to this email and he should be in contact next week. If you'd prefer to be contacted by telephone could you please provide us with a telephone number.

I hope this has been of some help. If I can be of any further assistance, please do not hesitate to let me know.

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Answer

Thank you for your inquiry.

I have to confirm that we have not tested this kit with bone marrow aspirate. If the bone marrow aspirate cell plasma is prepared cell free as a clear liquid we believe could be suitable for this sample type. But this is not covered by our guarantee since this was not yet tested.

We would recommend a starting dilution of 1:200 to 1:2000 for these kind of samples. Please note that this recommendation should only be used as a starting point and the optimal dilution will need to be determined empirically by you based on researched literature and your knowledge of these particular samples.

I am sorry I could not be of more help on this occasion and hope this information is nevertheless helpful.

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Answer

This kit has been successfully tested and validated with EDTA, citrate, and heparin plasma. Below are some general tips for sample preparation that will hopefully be useful for you:



TIPS FOR SAMPLE PREPARATION

How do I make conditioned media for a sample?

For conditioned medium, it is better to prepare serum free or low serum medium since some sera also contains cytokines. At day 0, seed 1 million cells in 100 mm tissue culture plate with complete medium. At day 3, remove medium and replace medium with 6-8 ml of low serum medium (e.g. medium containing 0.2% calf serum). At day 5, medium is collected into 15 ml tube. Spin at 2,000 rpm in centrifuge at 4 C for 10 minutes. Save the supernatant. Transfer the supernatant into 1.5 ml eppendorf tubes. Store supernatant at –80 C until experiment. The sample can be stored at this way for at least one year.

How do I make preparation of blood and serum samples?

For plasma:

Collect blood into EDTA or Sodium heparin tube

Spin 10 minutes at 3000 rpm

Aliquot into small tubes and store at –800C until use.

For serum:

Collect blood into tube without additive

Keep at room temperature for 20 minutes

Spin 10 minutes at 3000 rpm

Aliquot into small tubes and store at –800C until use.



How do I prepare urine samples?

Collect samples without adding stabilizers. Spin down the samples hard (eg. 10000 x g for 1min or 5000 x g for 2 min). Aliquot, quick freeze in dry ice/methanol bath, and store at –800C until use.



Please do not hesitate to contact us for further assistance.

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Question
Answer

Thank you for contacting us. The lab let me know the following: Our recommended human serum dilution for ab100628 is 200-2,000 fold.  Please note that this recommendation should only be used as a starting point and the optimal dilution will need to be determined empirically by the customer based on researched literature and knowledge of their particular samples. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.  

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Answer

Désolé pour la confusion à propos des vitesses de centrifugation. La préparation des échantillons plasma ou sérum doit se faire à 3000g. Le nombre de rotation par minute sera déterminé en fonction de la taille du rotor de la centrifugeuse utilisée. J'espère que ces informations vous seront utiles. N'hésitez pas à nous contacter de nouveau si vous avez d'autres questions.

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Answer

Merci de nous avoir contactés. J'ai reçu les informations suivantes concernant les kits ELISA CD40L et PF4 : ab99991 doit être utilisé avec des échantillons de plasma traités EDTA ou héparine. L'utilisation d'autres anticoagulants n'est pas recommandée. ab100628 peut être utilisé avec des échantillons de plasma traités EDTA, héparine ou citrate. Concernant les conditions de stockage, le laboratoire n'a pas d'information spécifique pour CD40 et PF4 dans le plasma. En règle générale, les protéines contenues dans le sérum ou le plasma sont stables à -80°C pour une durée d'environ un an. Préparation recommandée des échantillons : - Plasma : Collecter le sang un tube d'EDTA ou d'héparine. Centrifuger à 3000g pendant 10 minutes. Aliquoter et stocker à -80°C - Sérum : Collecter le sang sans additif. Maintenir à température ambiante pendant 20 minutes. Centrifuger à 3000g pendant 10 minutes. Aliquoter et stocker à -80°C. ------------------------ ab99991 should only be used with EDTA-plasma or heparin-plasma. Use of other anti-coagulants is not recommended. ab100628 can be used with EDA-plasma, heparin-plasma or citrate-plasma. FYI, unless otherwise stated in the Introduction Section of the manual, if the kit is compatible with plasma, it will be compatible with heparin, EDTA, and citrate anti-coagulants. As far as plasma sample storage, I’m not aware of a stability study specifically for CD40 ligand and Platelet Factor 4 in plasma, but in general most protein should be stable in serum or plasma for at least a year if stored at -80ºC. For Plasma : Collect blood into EDTA or Sodium heparin tube Spin 10 minutes at 3000g Aliquot into small tubes and store at –80°C until use. For serum: Collect blood into tube without additive Keep at room temperature for 20 minutes Spin 10 minutes at 3000g Aliquot into small tubes and store at –80°C until use.

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