製品の概要

  • 製品名Anti-Calnexin antibody [AF18]
    Calnexin 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [AF18] to Calnexin
  • 特異性Recognizes ER membrane, mitochondria and cis-Golgi
  • アプリケーション適用あり: Flow Cyt, ICC/IF, IP, WB, IHC-Pmore details
  • 種交差性
    交差種: Mouse, Human
  • 免疫原

    Focus cells (human hepatoma cell line)

  • ポジティブ・コントロール
    • This antibody gave a positive signal in the following, in WB - HeLa whole cell lysate, A431, HEK293; in IF - HeLa cells; in IHC - human liver carcinoma; in FC - HeLa.

製品の特性

  • 製品の状態Liquid
  • 保存方法Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • バッファーpH: 7.4
    Preservative: 0.02% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • 精製度IgG fraction
  • ポリ/モノモノクローナル
  • クローン名AF18
  • アイソタイプIgG1
  • 研究分野

アプリケーション

Our Abpromise guarantee covers the use of ab31290 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
Flow Cyt Use 1µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
ICC/IF Use a concentration of 1 µg/ml.
IP Use at an assay dependent concentration.
WB Use a concentration of 5 µg/ml. Detects a band of approximately 90 kDa (predicted molecular weight: 67 kDa).
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

ターゲット情報

  • 機能Calcium-binding protein that interacts with newly synthesized glycoproteins in the endoplasmic reticulum. It may act in assisting protein assembly and/or in the retention within the ER of unassembled protein subunits. It seems to play a major role in the quality control apparatus of the ER by the retention of incorrectly folded proteins.
  • 配列類似性Belongs to the calreticulin family.
  • 細胞内局在Endoplasmic reticulum membrane. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
  • Information by UniProt
  • 参照データベース
  • 別名
    • Calnexin antibody
    • CALX_HUMAN antibody
    • CANX antibody
    • CNX antibody
    • FLJ26570 antibody
    • Histocompatibility complex class I antigen binding protein p88 antibody
    • IP90 antibody
    • Major histocompatibility complex class I antigen-binding protein p88 antibody
    • p90 antibody
    see all

Anti-Calnexin antibody [AF18] 画像

  • Immunofluorescence staining of calnexin in HUVEC cells with ab31290. The samples were blocked with 1% BSA in PBS and the nuclei are stained with DAPI. FAM5C was also immuostained (red). The fluorescent signals were visualized with a laser scanning confocal microscope.

  • IHC image of Calnexin staining in human liver carcinoma FFPE section, performed on a 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 ab31290, 1µg/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.
  • All lanes : Anti-Calnexin antibody [AF18] (ab31290) at 5 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate (ab27252)
    Lane 2 : A431 (Human) Whole Cell Lysate (ab52255)
    Lane 3 : HEK293 (Human) Whole Cell Lysate (ab52256)

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/50000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 67 kDa
    Observed band size : 90 kDa (why is the actual band size different from the predicted?)


    Exposure time : 12 minutes

    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab31290 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406

  • ICC/IF image of ab31290 stained human HeLa cells. The cells were PFA fixed (10 min) and incubated with the antibody (ab31290, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used to quench autofluorescence.  5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue).  Alexa Fluor® 594 WGA was used to label plasma membranes (red).

  • Overlay histogram showing HeLa cells stained with ab31290 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab31290, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

Anti-Calnexin antibody [AF18] (ab31290) 使用論文

This product has been referenced in:
  • Noy PJ  et al. TspanC8 Tetraspanins and A Disintegrin and Metalloprotease 10 (ADAM10) Interact via Their Extracellular Regions: EVIDENCE FOR DISTINCT BINDING MECHANISMS FOR DIFFERENT TspanC8 PROTEINS. J Biol Chem 291:3145-57 (2016). Read more (PubMed: 26668317) »
  • Michailidou I  et al. Complement C3 on microglial clusters in multiple sclerosis occur in chronic but not acute disease: Implication for disease pathogenesis. Glia N/A:N/A (2016). IHC-P ; Human . Read more (PubMed: 27778395) »

See all 33 Publications for this product

Product Wall

Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (Mouse Embyronic Fibroblast)
Permeabilization Yes - 0.5% TX-100
Specification Mouse Embyronic Fibroblast
Blocking step Serum as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 4°C
Fixative Paraformaldehyde
Username

Ms. Samantha O'Hara

Verified customer

投稿 Oct 30 2015

Application Immunocytochemistry/ Immunofluorescence
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Sample Human Cell (fibroblasts)
Specification fibroblasts
Permeabilization Yes - 0.3% Triton X-100 in blocking solution
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

投稿 Mar 24 2014

Thank you for your call today.

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I am sorry that this antibody did not perform as stated on the datasheet. If payment has already been made on the original order and you wish to receive a refund, please ask y...

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Thank you for contacting us and your interest in our products. I have found a reference to Calnexin being used as a loading control of microsomal fractions (link below) and although i have not found reference to it being used in thyroid tissue specifi...

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Thanks for your email. I trust you received my email yesterday which noted I have requested a free vial of ab75801 be shipped to you for your customer. I have requested a confirmation be sent as soon as possible. Please let me know if you need anything...

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Sorry for the late reply. Our office was closed on Friday celebrate the Thanksgiving Day holiday. I have initiated a request for one vial of ab75801 to be sent as a replacement. Please let me know if you have any further questions.

Thanks for your reply. I would be happy to set a replacement of ab75801 for you. I'm sorry but I forgot to mention that I will need to locate your order for ab31290 so I can process your request. When you have a chance, please forward me the purchase o...

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Thanks for your reply and for sending the image. I agree that there are differencees between the 2 images of ER.  Interestingly, there is an ICC/IF Abreview for ab31290 which shows staining of H1299 cells which looks more similar to the staining you ar...

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Thank you for your enquiry and for including your protocol and image. Do you have an image of ER staining that you expect to see that you could forward to me? Thanks for the additional information!

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (H1299)
Specification H1299
Fixative Paraformaldehyde
Permeabilization Yes - 0.1% Triton X100
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Username

Abcam user community

Verified customer

投稿 Nov 17 2010

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