製品の概要

  • 製品名
    Human TGF beta 1 ELISA Kit
    TGF beta 1 キット 製品一覧
  • 検出方法
    Colorimetric
  • 再現性
    Intra-Assay(同時再現性)
    サンプル N 平均値 SD CV%
    Overall < 10%
    Inter-Assay(日差再現性)
    サンプル N 平均値 SD CV%
    Overall < 12%
  • 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
    < 80 pg/ml
  • 検出範囲
    0.082 ng/ml - 60 ng/ml
  • 添加回収試験

    96 %

    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 97.87 85% - 104%
    Serum 94.46 82% - 102%
    Plasma 95.78 93% - 103%

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

    Human TGF beta 1 ELISA (Enzyme-Linked Immunosorbent Assay) kit (ab100647) is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of Human TGF beta 1 in serum, plasma and cell culture supernatants.


    This assay employs an antibody specific for Human TGF beta 1 coated on a 96-well plate. Standards and samples are pipetted into the wells and TGF beta 1 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-Human TGF beta 1 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 TGF beta 1 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.


    Ab100647 was reformulated on 31st May 2018 with new capture and detector antibodies that allows to increase the sensitivity to human TGF beta 1.


    Please note that as a consequence of this change, the procedure and protocol have slightly changed and we encourage you to review the protocol steps before starting your experiments

  • 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
    500X HRP-Streptavidin Concentrate 1 x 200µl
    5X Assay Diluent 1 x 15ml
    Biotinylated anti-Human TGF beta 1 (lyophilized) 2 vials
    Recombinant Human TGF beta 1 Standard (lyophilized) 2 vials
    Stop Solution 1 x 8ml
    TGF beta 1 Microplate (12 x 8 wells) 1 unit
    TMB One-Step Substrate Reagent 1 x 12ml
  • 研究分野
  • 機能
    Multifunctional protein that controls proliferation, differentiation and other functions in many cell types. Many cells synthesize TGFB1 and have specific receptors for it. It positively and negatively regulates many other growth factors. It plays an important role in bone remodeling as it is a potent stimulator of osteoblastic bone formation, causing chemotaxis, proliferation and differentiation in committed osteoblasts.
  • 組織特異性
    Highly expressed in bone. Abundantly expressed in articular cartilage and chondrocytes and is increased in osteoarthritis (OA). Co-localizes with ASPN in chondrocytes within OA lesions of articular cartilage.
  • 関連疾患
    Defects in TGFB1 are the cause of Camurati-Engelmann disease (CE) [MIM:131300]; also known as progressive diaphyseal dysplasia 1 (DPD1). CE is an autosomal dominant disorder characterized by hyperostosis and sclerosis of the diaphyses of long bones. The disease typically presents in early childhood with pain, muscular weakness and waddling gait, and in some cases other features such as exophthalmos, facial paralysis, hearing difficulties and loss of vision.
  • 配列類似性
    Belongs to the TGF-beta family.
  • 翻訳後修飾
    Glycosylated.
    The precursor is cleaved into mature TGF-beta-1 and LAP, which remains non-covalently linked to mature TGF-beta-1 rendering it inactive.
  • 細胞内局在
    Secreted > extracellular space > extracellular matrix.
  • Information by UniProt
  • 別名
    • Camurati-Engelmann disease
    • CED
    • DPD1
    • LAP
    • Latency-associated peptide
    • TGF beta 1 protein
    • TGF-beta-1
    • TGFB
    • tgfb1
    • TGFB1_HUMAN
    • TGFbeta
    • Transforming growth factor beta 1
    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 ab100647 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.

プロトコール

参考文献

This product has been referenced in:
  • Corcelli M  et al. Neuroprotection of the hypoxic-ischemic mouse brain by human CD117+CD90+CD105+amniotic fluid stem cells. Sci Rep 8:2425 (2018). Read more (PubMed: 29402914) »
  • Wang X  et al. Cancer-associated fibroblasts induce epithelial-mesenchymal transition through secreted cytokines in endometrial cancer cells. Oncol Lett 15:5694-5702 (2018). Read more (PubMed: 29563996) »
See all 9 Publications for this product

レビューと Q&A

1-10 of 15 Abreviews or Q&A

TGFb human macrophage and DC secretion

Excellent Excellent 5/5 (Ease of Use)
Abreviews
TGFb human macrophage & DC secretion in culture for 3, isolated by negative selection.
The assay is easy to use and takes about 5.30hr.
Good specificity as shown by TGFb secretion by different macrophages subtyes & DC.
M0 = MCSF M1 = MCSF+IFNg M2 = MCSF+IL4 DC = GMCSF+IL4.

Abcam user community

Verified customer

投稿 May 02 2014

human TGF beta 1 detection in M6 cells

Excellent Good 4/5 (Ease of Use)
Abreviews
The Kit works without any remarkable problem.
It isn't a very consuming time protocol.
As control we use mouse cells supernatant and the result was very low.

Abcam user community

Verified customer

投稿 Oct 23 2013

Question
Answer

Thank you for contacting us. Yes, you can store the HCl and the Urea/Acetic acid for later use, you do not have to use the diluted versions all at once. Please let me know if you have any further questions.

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Answer



Avec ab100647, nous recommandons d’activer vos échantillons en faisant une extraction à l'acide, voir page 9 -10 du protocole.

Il est fortement recommandé d’effectuer cette l'extraction à l'acide car la TGFb actif naturellement sécrétée par les cellules est très peu abondante dans des échantillons normaux.

Selon nos données sur des échantillons normaux de sérum humain, la concentration apparente de TGF bêta 1 sans activation était d'environ 100 pg / ml. La valeur de la DO était assez faible et proche de la valeur DO pour les échantillons blancs. Cependant, après extraction, nous avons eu environ 2000 pg / ml.

Ce kit peut détecter entre 0.082 ng/ml - 60 ng/ml de TGF beta 1 mais il semblerait que sans l’activation du TGFb, les valeurs seraient plutôt vers l'extrémité inférieure de cette gamme. Je vous recommanderais plutôt d’utiliser TGF beta 1 Human ELISA Kit (ab108912). Ce kit ne suggère aucune extraction à l'acide et à une meilleure sensibilité de 0.031 ng/ml - 2 ng/ml.

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Answer

Both antibodies are produced in Rat and monoclonal.

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Answer

The laboratory that developed the ELISA did not send data showing results +/- acid treatment but did send the following summary. Please let me know if you still have questions or concerns.

It is strongly recommended that the customer performs the acid extraction as free/active TGF-beta 1 is likely innately very low in abundance in most normal samples. If however the customer’s samples are somehow treated and/or diseased with expected elevated levels of active TGF-beta 1, then she would not necessarily have to. In brief:

No Extraction – the kit will only detect the innately active TGF-beta 1. (i.e. TGF-beta 1 which is already free, not bound). The bound latent complex form will not be detected.

Extraction – the kit will detect both the innately active TGF-beta 1 and the now released active TGF-beta 1 (so total TGF-beta 1).

According to our data on normal human serum samples, the apparent concentration of TGF beta 1 without activation is around 100 pg/ml. The OD value was pretty low with a 2-fold dilution – close to background. After extraction, we got around 2000 pg/ml. So there is a significant difference.

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Answer

Thank you for contacting Abcam regarding ab100647.

I have received some further information from the lab regarding this issue and I wanted to share this with you.

The appearance of Assay Diluent B can vary by batch but a little cloudiness or precipitate is not something to be concerned with as it does contain protein and other solutes.While the solution appearance may look visibly unappealing, there is nothing based on our testing and customer feedback that says the assay or more importantly the data would be negatively affected. I would encourage the customer to bring the solution up to room temperature, mix thoroughly and dilute to the 1X working solution to help clarify the solution and incorporate any protein aggregates.

I hope that this information is helpful. Please let me know if you have any questions or there are other ways that Abcam may help you meet your research goals.

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Answer

Thank you again for your call yesterday, and I just want to follow-up to make sure your experimentwent smoothly.

Please keep me updated about the results using the modified diluent, and let me know if you have any questions or if there is anything else that we can do for you.

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Answer

Thank you for calling Abcam earlier today.
I have talked to the lab and they have told me that Assay Diluent B does contain some protein and solutes so it would not be uncommon to see a little cloudiness or floating unsolubilized protein aggregates (especially after storing at low temperatures and/or for a long period of time). Most of the time bringing the solution up to room temperature, mixing it thoroughly and diluting to the 1X working concentration will help clarify the solution and solubilize any protein aggregates or precipitates. Centrifuging and filtering the solution can also help but usually doing so is not needed.
Please let me know if there is anything else I can help you with.

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Answer

Thank yhou for your enquiry.

I have copied some information belowproviding tips for sample preparation when using this kit, which includes human serum prepration.

I hope this will be helpful to you. if you have any further questions, please do not hesitate to contact me.


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 -80oC until experiment. The sample can be stored at this way for at least one year.

For plasma:

Collect blood into EDTA or Sodium heparin tube
Spin 10 minutes at 3000 rpm
Aliquot into small tubes and store at -80oC 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 –80oC until use.

How do I prepare urine samples?

Collect samples without adding stabilizers. Spin down the samples hard (eg. 10000 x g for 1 min or 5000 x g for 2 min). Aliquot, quick freeze in dry ice/methanol bath, and store at -80oC until use.


How do I prepare cell or tissue lysates for use on the cytokine arrays?

The cell or tissue lysates for use with this kit can be prepared using most conventional methods, e.g. homogenization of cell or tissue in Lysis Buffer. We supply a compatible lysis buffer in most of our kits. Other general low-salt lysis buffers can be used with the following caveats:

1) Avoid using SDS or other strongly denaturing detergents. In general, non-ionic
detergents, such as Triton X-100 or NP-40 are best, although zwitterionic detergents,
such as CHAPS, or mild ionic detergents, such as sodium deoxycholate will work.

2) Use no more than 2% v/v total detergent

3) Avoid the use of sodium azide

4) Avoid reducing agents, such as dithiothreitol or mercaptoethanols

In general, we strongly recommend that you add some type of protease inhibitor cocktail to the lysis buffer prior to homogenization. Most general biochemical supply companies stock a wide variety of these products. Since susceptibility to proteolytic cleavage and the type of proteases present in the lysate vary, we do not recommend a specific product. Instead, your choice of which combination of protease inhibitors to use should be based upon a literature search for your protein(s) of interest and/or tissue or cell type. Phosphatase inhibitors may be used, but are not necessary unless the antibodies used in the kit specifically recognize phosphorylated (activated) forms of the protein.


Choices of the method for lysis and homogenization include glass-bead smash douncing, freeze-thaw, sonication and crushing frozen tissue with a mortar and pestle, or even a combination of these. There is no one best method for all sample types, but some are better than others for some sample types. Your choice of method should be made following a brief search of the literature to see how samples similar to yours have been prepared in previous investigations.

After homogenization, spin down the lysates to remove cell/tissue debris (5 min @ 10000 x g or 10 min @ 5000 x g) and save the supernatant. Lysates should be frozen as soon as possible, and stored at -20oC (or -80oC, if possible). Spin them again before incubating with the antibody array. Determine the protein concentration of your lysates (for example the bicinchoninic acid (BCA) assay) and normalize the volume of each sample used to deliver the same amount of total protein for each assay.

Since different cells and tissues may contain different amounts of protein, as starting point, we suggest using 500 uL of lysis buffer per 1x106 cells or 10 mg tissue. You may have to adjust this based upon your results. Your target for total protein concentration of the homogenate should be at least 1000 ug/mL, but 2000 ug/mL would be better.

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

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