製品の概要

  • 製品名Anti-Calnexin antibody
    Calnexin 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to Calnexin
  • 特異性Recognizes ER membrane, mitochondria and cis-Golgi
  • アプリケーション適用あり: WB, ICC/IF, IHC-P, IP, IHC-Frmore details
  • 種交差性
    交差種: Mouse, Rat, Human, Marmoset (common)
    交差が予測される動物種: Dog
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 550 to the C-terminus of Human Calnexin.

    (Peptide available as ab23379.)

  • ポジティブ・コントロール
    • ab22595 gave a positive result in the following Human Whole Cell Lysates: Hela, MCF-7 Mouse Whole Cell Lystates: NIH 3T3, MEF1 Mouse Tissue Lysates: Brain, Liver, Heart, Kidney, Pancreas, Testis, Skeletal muscle, Spinal cord, Ovary Rat Whole Cell Lysates: PC12, Rat Tissue Lysates: Brain, Liver, Heart, Kidney

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. Store In the Dark.
  • バッファーPreservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • 精製度Immunogen affinity purified
  • ポリ/モノポリクローナル
  • アイソタイプIgG
  • 研究分野

アプリケーション

Our Abpromise guarantee covers the use of ab22595 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
WB Use a concentration of 1 µg/ml. Detects a band of approximately 75 kDa (predicted molecular weight: 90 kDa).
ICC/IF Use a concentration of 5 µg/ml.
IHC-P Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.

ターゲット情報

Anti-Calnexin antibody 画像

  • All lanes : Anti-Calnexin antibody (ab22595) at 1/250 dilution

    Lane 1 : NIH 3T3 whole cell lysate (ab7179)
    Lane 2 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate (ab46770)
    Lane 3 : Brain (Mouse) Tissue Lysate (ab27253)
    Lane 4 : Liver (Mouse) Tissue Lysate (ab7935)
    Lane 5 : Heart (Mouse) Tissue Lysate (ab27255)
    Lane 6 : Kidney (Mouse) Tissue Lysate (ab27254)
    Lane 7 : Mouse pancreas tissue lysate - total protein (ab29363)
    Lane 8 : Testis (Mouse) Tissue Lysate - normal tissue (ab4027)
    Lane 9 : Mouse skeletal muscle tissue lysate - total protein (ab29711)
    Lane 10 : Spinal Cord (Mouse) Tissue Lysate (ab50253)
    Lane 11 : Ovary (Mouse) Tissue Lysate (ab35808)
    Lane 12 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate (ab50957)
    Lane 13 : Brain (Rat) Tissue Lysate (ab7942)
    Lane 14 : Liver (Rat) Tissue Lysate (ab27256)
    Lane 15 : Heart (Rat) Tissue Lysate (ab7940)
    Lane 16 : Kidney (Rat) Whole Cell Lysate - normal tissue (ab29480)

    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 : 90 kDa
    Observed band size : 80 kDa (why is the actual band size different from the predicted?)
  • ICC/IF image of ab22595 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab22595, 5µg/ml) for 1h at room temperature. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h (green). Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 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).

  • Kidney cortex using ab22595 shows clear cytoplasmic staining patterns. The visceral cells of the Glomerular tuft ( podocytes ) are strongly stained (indicated by red arrowheads). Distal convoluted tubular cells are generally moderately positive (with exceptions that are strongly positive). However, most of the cells that line the Proximal Convoluted Tubules (indicated by green arrowheads) are strongly positive.

    See Abreview

  • All lanes : Anti-Calnexin antibody (ab22595) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line)
    Lane 2 : U2OS Whole Cell Lysate
    Lane 3 : MCF-7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
    Lane 4 : HeLa (Human epithelial carcinoma cell line) with Human Calnexin - ER membrane marker peptide (ab23379) at 1 µg/ml
    Lane 5 : U2OS Whole Cell Lysate with Human Calnexin - ER membrane marker peptide (ab23379) at 1 µg/ml
    Lane 6 : MCF-7 (Human breast adenocarcinoma cell line) Whole Cell Lysate with Human Calnexin - ER membrane marker peptide (ab23379) at 1 µg/ml

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG (Alexa Fluor® 680) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size : 90 kDa
    Observed band size : 75 kDa (why is the actual band size different from the predicted?)

    Recent batches of ab22595 (AP217379 and AP151845) detect a band of ~ 75 kDa in Hela, U2OS and MCF-7 lysates. This band is completely blocked by the immunizing peptide so we believe this represents Calnexin. Moreoever, a band of the same size is detected by other Calnexin antibodies tested.

  • ICC/IF image of ab22595 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 (ab22595, 5µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-rabbit IgG - H&L, pre-adsorbed (ab96899) 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.

  • Calnexin - ER membrane marker was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to Calnexin - ER membrane marker and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab22595.
    Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution.
    Band: 80kDa: Calnexin - ER membrane marker.

Anti-Calnexin antibody (ab22595) 使用論文

This product has been referenced in:
  • Shen J  et al. NMDA receptors participate in the progression of diabetic kidney disease by decreasing Cdc42-GTP activation in podocytes. J Pathol 240:149-60 (2016). Read more (PubMed: 27338016) »
  • Dametto P  et al. Neurodegeneration and unfolded-protein response in mice expressing a membrane-tethered flexible tail of PrP. PLoS One 10:e0117412 (2015). IF ; Mouse . Read more (PubMed: 25658480) »

See all 29 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (primary keratinocyte)
Permeabilization Yes - 0.4% TritonX100
Specification primary keratinocyte
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 12% · Temperature: 25°C
Fixative Paraformaldehyde
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投稿 Sep 05 2016

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Application Immunohistochemistry (Frozen sections)
Sample Human Tissue sections (face skin)
Permeabilization Yes - 0.4% TritonX100
Specification face skin
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative Acetone
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投稿 Sep 01 2016

Application Immunocytochemistry
Sample Rat Cultured Cells (primary Hippocampal neuron)
Permeabilization Yes - 0.2% Triton X-100 in PBS
Specification primary Hippocampal neuron
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 23°C
Fixative Paraformaldehyde
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投稿 Mar 11 2016

Application Western blot
Sample Rat Cell lysate - whole cell (RBL-2H3 cell)
Gel Running Conditions Reduced Denaturing
Loading amount 10 µg
Specification RBL-2H3 cell
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
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投稿 Sep 11 2015

Application Western blot
Sample Human Cell lysate - whole cell (HEK293T cells)
Gel Running Conditions Reduced Denaturing
Loading amount 10 µg
Specification HEK293T cells
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
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投稿 Jun 19 2015

Application Western blot
Loading amount 10000 cells
Gel Running Conditions Reduced Denaturing (4-12% gradient)
Sample Human Cell lysate - whole cell (Huh7 liver)
Specification Huh7 liver
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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投稿 Feb 05 2015

Application Western blot
Loading amount 10 µg
Gel Running Conditions Reduced Denaturing (16%)
Sample Mouse Cell lysate - whole cell (Primary bone marrow derived macrophage)
Specification Primary bone marrow derived macrophage
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 20°C
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投稿 Oct 16 2014

Application Immunocytochemistry/ Immunofluorescence
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Sample Human Cell (ovarian)
Specification ovarian
Permeabilization Yes - 0.1% triton X for 5 min at RT
Fixative Paraformaldehyde
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投稿 Aug 25 2014

Application Western blot
Loading amount 40 µg
Gel Running Conditions Reduced Non-Denaturing (Native) (12)
Sample Rat Tissue lysate - other (membrane fraction)
Specification membrane fraction
Blocking step Milk as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 5% · Temperature: 23°C
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投稿 Nov 18 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Loading amount 3 µg
Gel Running Conditions Reduced Denaturing (10%)
Sample Mouse Tissue lysate - other (microsomes)
Specification microsomes
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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投稿 Aug 29 2013

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