使用敲除细胞株进行验证

Anti-Calreticulin抗体- ER Marker (ab39818)

概述

  • 产品名称Anti-Calreticulin抗体- ER Marker
    参阅全部 Calreticulin 一抗
  • 描述
    兔多克隆抗体to Calreticulin - ER Marker
  • 经测试应用适用于: IHC-P, WB, ICC/IFmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
    预测可用于: Rabbit, Chinese Hamster
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 250 - 350 of Human Calreticulin.

    (Peptide available as ab39895.)

  • 阳性对照
    • WB: Jurkat, HepG2, A431, PC12, NIH3T3 and MEF1 whole cell lysates.

性能

  • 形式Liquid
  • 存放说明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.
  • 存储溶液Preservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • 纯度Immunogen affinity purified
  • 克隆多克隆
  • 同种型IgG
  • 研究领域

相关产品

应用

Our Abpromise guarantee covers the use of ab39818 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
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 55 kDa (predicted molecular weight: 48 kDa).
ICC/IF Use a concentration of 1 µg/ml.

靶标

  • 功能Molecular calcium-binding chaperone promoting folding, oligomeric assembly and quality control in the ER via the calreticulin/calnexin cycle. This lectin interacts transiently with almost all of the monoglucosylated glycoproteins that are synthesized in the ER. Interacts with the DNA-binding domain of NR3C1 and mediates its nuclear export.
  • 序列相似性Belongs to the calreticulin family.
  • 结构域Can be divided into a N-terminal globular domain, a proline-rich P-domain forming an elongated arm-like structure and a C-terminal acidic domain. The P-domain binds one molecule of calcium with high affinity, whereas the acidic C-domain binds multiple calcium ions with low affinity.
    The interaction with glycans occurs through a binding site in the globular lectin domain.
    The zinc binding sites are localized to the N-domain.
    Associates with PDIA3 through the tip of the extended arm formed by the P-domain.
  • 细胞定位Endoplasmic reticulum lumen. Cytoplasm > cytosol. Secreted > extracellular space > extracellular matrix. Cell surface. Also found in cell surface (T cells), cytosol and extracellular matrix. Associated with the lytic granules in the cytolytic T-lymphocytes.
  • Information by UniProt
  • 数据库链接
  • 别名
    • Autoantigen RO antibody
    • CALR antibody
    • CALR protein antibody
    • CALR_HUMAN antibody
    • Calregulin antibody
    • Calreticulin antibody
    • cC1qR antibody
    • CRP55 antibody
    • CRT antibody
    • CRTC antibody
    • Endoplasmic reticulum resident protein 60 antibody
    • Epididymis secretory sperm binding protein Li 99n antibody
    • ERp60 antibody
    • FLJ26680 antibody
    • grp60 antibody
    • HACBP antibody
    • HEL S 99n antibody
    • RO antibody
    • Sicca syndrome antigen A antibody
    • SSA antibody
    see all

Anti-Calreticulin antibody - ER Marker 图像



  • Predicted band size : 48 kDa
    Additional bands at : 55 kDa. We are unsure as to the identity of these extra bands.

    Lane 1: Wild-type HAP1 cell lysate (20 µg)

    Lane 2: Calreticulin knockout HAP1 cell lysate (20 µg)

    Lane 3: HeLa cell lysate (20 µg)

    Lane 4: NIH3T3 cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab39818 observed at 55 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab39818 was shown to specifically react with Calreticulin when Calreticulin knockout samples were used. Wild-type and Calreticulin knockout samples were subjected to SDS-PAGE. ab39818 at a concentration of 1 μg/ml and ab8245 (loading control to GAPDH) diluted to 1/10000 were both incubated overnight at 4°C. Blots were developed with goat anti-rabbit IgG (H + L) and goat anti-mouse IgG (H + L) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • All lanes : Anti-Calreticulin antibody - ER Marker (ab39818) at 1 µg/ml

    Lane 1 : Jurkat whole cell lysate (ab7899)
    Lane 2 : HepG2 whole cell lysate (ab7900)
    Lane 3 : A431 whole cell lysate (ab7909)

    Lysates/proteins at 10 µg per lane.

    Secondary
    IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size : 48 kDa
    Observed band size : 55 kDa (why is the actual band size different from the predicted?)
    The predicted band size is 48kDa based on Swiss-prot data, however a band of 55kDa is consistent with that observed in other commercially available antibodies to Calreticulin. This difference in size is likely to be the result of post-translational modification.
    ICC/IF image of ab39818 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab39818, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).

  • IHC image of Calreticulin 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 ab39818, 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-Calreticulin antibody - ER Marker (ab39818) at 1 µg/ml

    Lane 1 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
    Lane 2 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 3 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Performed under reducing conditions.

    Predicted band size : 48 kDa
    Observed band size : 55 kDa (why is the actual band size different from the predicted?)


    Exposure time : 8 minutes

  • Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 48 kDa


    Exposure time : 30 seconds

Anti-Calreticulin antibody - ER Marker (ab39818)参考文献

ab39818 has not yet been referenced specifically in any publications.

Product Wall

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Fixative Methanol
Permeabilization No
Username

Dr. Kirk McManus

Verified customer

提交于 May 12 2008

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