製品の概要

  • 製品名
    Human IGFBP2 ELISA Kit
    IGFBP2 キット 製品一覧
  • 検出方法
    Colorimetric
  • サンプルの種類
    Cell culture supernatant, Serum, Plasma
  • アッセイタイプ
    Sandwich (quantitative)
  • 検出感度
    < 20 pg/ml
  • 検出範囲
    8.23 pg/ml - 6000 pg/ml
  • 添加回収試験

    97 %

    特定サンプルでの回収試験
    サンプルの種類 平均 % 測定範囲
    Cell culture supernatant 100.25 86% - 105%
    Serum 95.77 84% - 103%
    Plasma 97.59 85% - 104%

  • ステップ
    Multiple steps standard assay
  • 種交差性
    交差種: Human
  • 製品の概要

    Abcam’s IGFBP2 Human ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of Human IGFBP2 in serum, plasma and cell culture supernatants.

    This assay employs an antibody specific for Human IGFBP2 coated on a 96-well plate. Standards and samples are pipetted into the wells and IGFBP2 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-Human IGFBP2 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 IGFBP2 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

  • 特記事項

    Optimization may be required with urine samples

  • アプリケーション
    適用あり: Sandwich ELISAmore details
  • 試験プラットフォーム
    Microplate

製品の特性

  • 保存方法
    Store at -20°C. Please refer to protocols.
  • 内容 1 x 96 tests
    20X Wash Buffer Concentrate 1 x 25ml
    300X HRP-Streptavidin Concentrate 1 x 200µl
    5X Assay Diluent B 1 x 15ml
    Assay Diluent A 1 x 30ml
    Biotinylated anti-Human IGFBP2 (lyophilized) 2 vials
    IGFBP2 Microplate (12 x 8 wells) 1 unit
    Recombinant Human IGFBP2 Standard (lyophilized) 2 vials
    Stop Solution 1 x 8ml
    TMB One-Step Substrate Reagent 1 x 12ml
  • 研究分野
  • 機能
    Inhibits IGF-mediated growth and developmental rates. IGF-binding proteins prolong the half-life of the IGFs and have been shown to either inhibit or stimulate the growth promoting effects of the IGFs on cell culture. They alter the interaction of IGFs with their cell surface receptors.
  • 配列類似性
    Contains 1 IGFBP N-terminal domain.
    Contains 1 thyroglobulin type-1 domain.
  • ドメイン
    The C-terminus is required for IGF-binding and growth inhibition.
  • 翻訳後修飾
    O-glycosylated.
  • 細胞内局在
    Secreted.
  • Information by UniProt
  • 別名
    • BP 2
    • BP2
    • IBP 2
    • IBP-2
    • IBP2
    • IBP2_HUMAN
    • IGF binding protein 2
    • IGF BP53
    • IGF-binding protein 2
    • IGF-BP53
    • IGFBP-2
    • IGFBP2
    • IGFBP53
    • Insulin Like Growth Factor Binding Protein 2
    • Insulin like growth factor binding protein 2 36kDa
    • Insulin like growth factor-binding protein 2 precursor
    • Insulin-like growth factor-binding protein 2
    see all
  • 参照データベース

アプリケーション

Our Abpromise guarantee covers the use of ab100540 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
Sandwich ELISA Use at an assay dependent concentration.

画像

  • Representative standard curve using ab100540

  • Representative standard curve using ab100540

プロトコール

参考文献

This product has been referenced in:
  • Chang YC  et al. The generation of induced pluripotent stem cells for macular degeneration as a drug screening platform: identification of curcumin as a protective agent for retinal pigment epithelial cells against oxidative stress. Front Aging Neurosci 6:191 (2014). Sandwich ELISA ; Human . Read more (PubMed: 25136316) »
  • Sunderic M  et al. Protein molecular forms of insulin-like growth factor binding protein-2 change with aging. Exp Gerontol 58C:154-158 (2014). Read more (PubMed: 25106097) »
See all 3 Publications for this product

レビューと Q&A

1-8 of 8 Abreviews or Q&A

Answer

Thank you and your customer for your inquiry.

Although we have not tested the Human ab100540 and ab100544ELISA kits with ascite fluid,these kits are very accommodating to many different samples types so the ascites will most likely work.

Please let your customer know that they needto use Assay Diluent B for this sample type.

I hope this information is helpful and wish your customer good luck with their experiments.

Read More

Answer

Thank you for contacting Abcam.

For both of these ELISA kits we do recommend that you make the wash buffer up fresh before every use and we haven't tested the long term stability of the buffer when diluted to 1X. It should be good if stored overnight at 4C, but any longer and we cannot guarantee that it will be useable.

If there is anything else I can help you with, please let me know.

Read More

Answer

Thank you for contacting us.

Because we carry over 80,000 products, it isn't feasible for us to keep small sample sizes of our products. Therefore, I am afraid I cannot send you a free sample of the IGFBP2 Human ELISA Kit ab100540.

Although I agree that on page 9 of the protocol the instruction says ' Add 100 µl of each standard [...] and sample into appropriate wells....', it also clearly refers to the Preparation of Reagents step 2 (page 7). Also, in the assay summary on page 4 it is also recommended to 'Prepare all reagents, samples and standards as instructed'.

This Preparation of Reagents step 2 ('Sample dilution: If your samples need to be diluted, Assay Diluent A (Item D) is used for dilution of serum/plasma samples...') refers to the common scientific practise to determine the optimal sample dilution. This dilution is required to ensure that the readings are within the standard curve range. Also, the table on page 12 (D. Linearity) indicates the linearity of dilution which refers to the predictability of sample recovery for known dilution factors in the desired assay range. This might give you an indication to what dilutions might be useful for your particular sample type.

I am sorry that I cannot be of more assistance on this occasion, but I hope this information is nevertheless helpful for you. Please do not hesitate to contact us again with any other questions.

Read More

Answer

I apologize for the delay in getting back to you in regards to this issue, and I appreciate your patience as I have been discussing this situation with my colleagues. I can understand your frustration and disappointment as the results have been unexpected with these kits, and we certainly want you to be satisfied with them. After looking through the data and discussing with scientists in the lab, we do have some additional suggestions, and though I realize you have previously troubleshooted we would greatly appreciate if you could review the information below and let me know if you would be willing to try again. We really want to get these kits to work for you, as the standard curves seem to be fine for each of the kits, and therefore the plasma samples themselves need further optimization. For the IGFBP2 kit ab100540, the increase in signal with dilution indicates a matrix effect with the sample. The dilutions that the lab recommends for this particular kit are between 1:200 and 1:2000, so 1:27 is probably not dilute enough in order to have accurate results. We would recommend trying a few dilutions between 1:200 and 1:2000 until a linear response is seen. CNTF can be a fairly low abundant protein, and so the readings might be pretty low. The lab recommends a 1:2 dilution for plasma used with this kit (don't use undiluted plasma as matrix effects can interfere with reaction), and so the current dilutions seem to be much too high. We would recommend trying a 1:2 dilution and some protocol alterations to increase the signal, such as leaving the incubation steps overnight at 4C with gentle rocking, and increasing the amount of biotinylated antibody or HRP-streptavidin if the signal is still weak (increase by 1.5 fold or so). You should still expect that the signal might be in the lower range of the standard curve, if the protein is being expressed at low levels. For the IGF1 kit ab100545, the average readings were around 6 ng/mL, which is not far from the 10-1000 ng/mL range that is generally found in normal healthy plasma. To reduce the volume that the plasma samples are diluted, you can alter the extraction protocol in step 3 by titrating the supernatant to pH 7-8 with NaOH solution instead of adding the 200 uL of Tris buffer. This should bring up the sample readings. Again, I understand that you have tried the kits a few times, but it does seem like the sample preparation can be optimized in order to produce the desired results. The standard curves are linear which indicates that the components are working correctly, but plasma samples often need to be optimized especially for these low-level molecules. I would appreciate if you could give me a call tomorrow so that we can further discuss the data and the next steps in this process. I look forward to hearing from you.

Read More

Answer

Thank you very much for the data. I am working with some of my colleagues to resolve this as quickly as possible, and I appreciate your patience and understanding! Please let me know if there is anything you need in the meantime.

Read More

Answer

Thank you for your reply. I have found the order with the additional kits that you mentioned. In order to process this refund, I will need a little bit more technical information. Have you tried all of these kits, or did you test 1 from each set and the remaining 7 are unused? Could you please send some data for the CNTF and IGFBP2 kits? I look forward to hearing from you. As this is a rather large refund, it may take a couple of days to complete your request, but we will try to finish this as soon as possible. Please let me know if you have any questions and I'll be happy to help.

Read More

Answer

Thank you for your reply, and I apologize that these kits did not work well for you. I did not find the attached summary of the test results, could you please send these again? We do fully guarantee these kits to work, so I have issued credit note *** for this order. The full purchase price will be refunded back to your credit card pending approval from our accounting department. If you have any questions about this, please contact our accounting department at us.credits@abcam.com. Again, I am sorry to hear about the poor performance of these kits. We have extensively troubleshooted for ab100545 so I am familar with that situation, but if you have any additional details about the trouble with ab100540 and ab99994 that may be beneficial for our investigation, please forward this information to me. I look forward to hearing from you. If there is anything else that we can do for you, please do let me know.

Read More

Answer

Thank you for your patient. I just had a response from my lab colleagues. They confirmed that all three kits were designed against the mature form of the protein (which is the full amino acid sequence minus the signalling peptide). As the epitope is located somewhere on the mature form, the detection antibodies will both recognise uncleaved and cleaved forms of the protein. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

登録