概述

  • 产品名称
  • 描述
    兔多克隆抗体to CETP
  • 宿主
    Rabbit
  • 经测试应用
    适用于: ICC/IF, IHC-P, WBmore details
  • 种属反应性
    与反应: Human
    预测可用于: Rabbit, Hamster, Non human primates
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 450 to the C-terminus of Human CETP.

    参阅Abcam的专有抗源政策 (Peptide available as ab71389.)

  • 阳性对照
    • Recombinant Human CETP protein (ab114408) can be used as a positive control in WB. This antibody gave a positive signal in the following whole cell lysates: HeLa, Jurkat, SK N BE, Y79 and HepG2.

性能

应用

Our Abpromise guarantee covers the use of ab51771 in the following tested applications.

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

应用 Ab评论 说明
ICC/IF Use a concentration of 5 µg/ml.
IHC-P Use a concentration of 5 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 70 kDa (predicted molecular weight: 55 kDa).

靶标

  • 功能
    Involved in the transfer of insoluble cholesteryl esters in the reverse transport of cholesterol.
  • 组织特异性
    Expressed by the liver and secreted in plasma.
  • 疾病相关
    Defects in CETP are a cause of hyperalphalipoproteinemia (HYPALIP) [MIM:143470]. Affected individuals show high levels of alpha-lipoprotein (high density lipoprotein/HDL).
    Defects in CETP are the cause of cholesteryl ester transfer protein deficiency (CETP deficiency) [MIM:607322]. This is an autosomal dominant condition associated with increased HDL cholesterol levels.
  • 序列相似性
    Belongs to the BPI/LBP/Plunc superfamily. BPI/LBP family.
  • 细胞定位
    Secreted > extracellular space.
  • Information by UniProt
  • 数据库链接
  • 别名
    • BPIFF antibody
    • CETP antibody
    • CETP_HUMAN antibody
    • Cholesteryl ester transfer protein antibody
    • Cholesteryl ester transfer protein plasma antibody
    • HDLCQ10 antibody
    • Lipid transfer protein I antibody
    see all

图片

  • All lanes : Anti-CETP antibody (ab51771) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 3 : SK N BE (Human neuroblastoma) Whole Cell Lysate
    Lane 4 : Y79 (Human retinoblastoma cell line) Whole Cell Lysate
    Lane 5 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size: 55 kDa
    Observed band size: 70 kDa
    why is the actual band size different from the predicted?


    Exposure time: 20 minutes


    CETP contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.
  • ICC/IF image of ab51771 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab51771, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899 Dylight 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • IHC image of ab51771 staining CETP in Human liver formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab51771, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

文献

This product has been referenced in:
  • van der Tuin SJL  et al. Lipopolysaccharide Lowers Cholesteryl Ester Transfer Protein by Activating F4/80+Clec4f+Vsig4+Ly6C- Kupffer Cell Subsets. J Am Heart Assoc 7:N/A (2018). Read more (PubMed: 29525783) »
See 1 Publication for this product

客户评价及客户问答

1-6 of 6 Abreviews or Q&A

Answer

Thank you for your email.
The protease inhibitors are added in the lysis buffer so when you do centrifugation there is no need of adding protease inhibitors in supernatant again.
Supernatant I mean the media when the cells are pelleted out. The same name (most importantly lysates) is also used when cells are lysed and debris are pelleted out.
Antibodies are recommended to be used at certain dilutions e.g. 1/100, 1/500 or 1/1000 dilutions however we recommend them storing them neat after aliquoting e.g. 10 ul per vial and storing them as recommended on the datasheet. Antibodies retains biological activity for longer duration when stored neat as compared to diluted one.
Our Hong Kong office deal with Indonesia orders. Please contact them at;
Unit 712, 7th Floor, Lakeside 1,
No. 8 Science Park West Avenue,
Hong Kong Science Park,
Pak Shek Kok,
New Territories,
Hong Kong
Orders:hk.orders@abcam.com
Technical:hk.technical@abcam.com
Tel: Tel: (852) 2603-6823
I hope this will help.

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Answer

Thank you very much for providing details.
Could you provide the details of distributor form whom this product was purchased? Are they official Abcam distributor? Please note, if they are not official then Abpromise guarantee will not be applied to this purchase which means you will not be entitled to refunds or free of charge replacements - we can however help with troubleshooting.
The further questions I have are
- Did you try a different antibody against the same target with same supernatant and lysates?
- Have you added loading buffer in supernatant as well as lysates?
- I would suggest adding protease inhibitors in the lysis buffer and trying the HepG2 lysates.
The antibody should have worked so the other reason of antibody being gone off is due to shipping temperature or due to delay in customs; this is the reason why we encourage customers to buy products from authorized distributors only because we assess their shipping and storage before making any distributorship deal.
I hope this information will nevertheless helpful. Should you have any other question please do not hesitate to contact me.

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Answer

Thank you for contacting us.
Could you clarify the problem? Are you concerned that antibody shows multiple bands? If yes could you answer the following questions;
- What is the order number and how the antibody was stored?
- Have you added any protease inhibitors in lysis buffer?
- What was the cell passage number and confluency percentage when lysates were made?
- Have you tried loading control and negative control?
- Could you provide an image with molecular weight ladder?
I will look forward to hearing from you soon.

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Answer

Thank you for contacting us.

This particular product ab51771 does not contain BSA when the antibody concentration is ≥1.00 mg/ml, which is the case for the lot we currently have in stock. BSA is added in the storage buffer of ab51771 when the concentration is below 1 mg/ml in order to increase the stability of the antibody.

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

Thank you for contacting us. We do not have any free samples of the liver tissue that was used to validate this antibody for IHC, but if you are interested in purchasing sections of other liver tissue, we have sections of paraffin-embedded human liver  (ab4348) and mouse liver (ab4607). Please do not hesitate to contact us if you need any more advice or information.  

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Answer

Thank you for contacting us. The conditions for the IHC validation are described briefly in the caption of the image on the online datasheet, which shows a stain of a formalin-fixed, paraffin-embedded human liver section. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab51771, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The other details of the protocol are standard. We recommend blocking non-specific staining with a 5% solution of normal serum from the species that the secondary antibody was derived from. The blocking step is typically 30-60 minutes at room temperature, followed by the primary antibody incubation. The wash solutions should contain 0.1% Triton X-100 or Tween-20. 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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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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