重组
RabMAb

Anti-Calmodulin抗体[EP799Y] (ab45689)

概述

  • 产品名称
    Anti-Calmodulin抗体[EP799Y]
    参阅全部 Calmodulin 一抗
  • 描述
    兔单克隆抗体[EP799Y] to Calmodulin
  • 经测试应用
    适用于: WB, IHC-P, ICC/IF, Flow Cyt, IP, IHC-Frmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) (C terminal)

  • 阳性对照
    • WB: C6, RAW264.7, NIH/3T3, HeLa, HCT 116 cell lysate. IHC-P: Human urinary bladder carcinoma and testis tissues. ICC/IF: MCF7 cells. Flow Cyt: MCF7 and NIH/3T3 cells. IP: Human skeletal muscle tissue lysate.
  • 常规说明

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

性能

应用

Our Abpromise guarantee covers the use of ab45689 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
WB 1/1000 - 1/10000. Detects a band of approximately 16 kDa (predicted molecular weight: 17 kDa).
IHC-P 1/2000.

Mouse and rat species reactivity is recommended based on WB results, we do not guarantee IHC for mouse or rat.

ICC/IF 1/100 - 1/250.
Flow Cyt 1/50 - 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IP 1/20 - 1/50.
IHC-Fr Use at an assay dependent concentration.

Mouse and rat species reactivity is recommended based on WB results, we do not guarantee IHC for mouse or rat.

靶标

图片

  • All lanes : Anti-Calmodulin antibody [EP799Y] (ab45689) at 1/10000 dilution

    Lane 1 : HeLa (human cervix adenocarcinoma) whole cell lysate
    Lane 2 : HCT 116 (human colorectal carcinoma) whole cell lysate

    Blocking peptides at 15 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size : 17 kDa

    Blocking and Diluting buffer: 5% NFDM/TBST

  • ab45689 immunoprecipitating Calmodulin. 10µg of cell lysate was incubated with primary antibody at a dilution of 1/20 and VeriBlot for IP secondary antibody (HRP) (ab131366) at a dilution of 1/10000.

    Lane 1: Human skeletal muscle lysate (10ug)
    Lane 2: Human skeletal muscle lysate
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab45689 in Human skeletal muscle lysate

  • ab45689 staining Calmodulin in human testis tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/2000. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

    Negative control 1: PBS in place of primary antibody.

  • ab45689 staining Calmodulin in NIH/3T3 (mouse embryo) cells by flow cytometry. Cells were fixed with 4% paraformaldehyde and the sample was incubated with the primary antibody at a dilution of 1/100. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody.

    Isoytype control: Rabbit monoclonal IgG (Black)

    Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)

  • ICC/IF image of ab45689 stained MCF7 cells. The cells were 100% methanol fixed (5 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 (ab45689, 1/1000 dilution) overnight at +4°C. 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) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • All lanes : Anti-Calmodulin antibody [EP799Y] (ab45689) at 1/10000 dilution

    Lane 1 : RAW264.7 (mouse abelson murine leukemia virus-induced tumor) whole cell lysate
    Lane 2 : C6 (rat glioma) whole cell lysate

    Lysates/proteins at 15 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size : 17 kDa

    Blocking and diluting buffer: 5% NFDM/TBST

  • ab45689 staining Calmodulin in Mouse brain tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with acetone and blocked with 10% Serum for 60 minutes at 4°C. Samples were incubated with primary antibody (1/100 in PBS + 0.1% Triton) for 12 hours at 4°C. A Alexa Fluor® 555-conjugated Goat anti-rabbit IgG polyclonal (1/400) was used as the secondary antibody.

    See Abreview

  • Ab45689 (1:250) staining human Calmodulin in human urinary bladder carcinoma by immunohistochemistry in paraffin embedded tissue.
  • Anti-Calmodulin antibody [EP799Y] (ab45689) at 1/5000 dilution + NIH 3T3 cell lysate (10ug/lane)

    Predicted band size : 17 kDa
    Observed band size : 16 kDa (why is the actual band size different from the predicted?)
  • Overlay histogram showing MCF-7 cells stained with ab45689 (red line). The cells were fixed with 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 (ab45689, 1/50 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (0.5µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a decreased signal in MCF-7 cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

文献

This product has been referenced in:
  • Burkard C  et al. Precision engineering for PRRSV resistance in pigs: Macrophages from genome edited pigs lacking CD163 SRCR5 domain are fully resistant to both PRRSV genotypes while maintaining biological function. PLoS Pathog 13:e1006206 (2017). Read more (PubMed: 28231264) »
  • Pehar M  et al. Changes in Protein Expression and Lysine Acetylation Induced by Decreased Glutathione Levels in Astrocytes. Mol Cell Proteomics 15:493-505 (2016). Read more (PubMed: 26486419) »

See all 20 Publications for this product

客户评价及客户问答

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (liver, hepatocytes)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
liver, hepatocytes
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

Abcam user community

Verified customer

提交于 Apr 26 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Drosophila melanogaster Tissue lysate - other (Head)
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
50 µg
Specification
Head
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

提交于 Mar 24 2017

Application
ELISA
Sample
Human Recombinant protein (Human Calmodulin full length protein)
Specification
Human Calmodulin full length protein
Type
Sandwich (Capture)
Username

Mr. Chang Su Jeon

Verified customer

提交于 Jul 27 2015

Application
Western blot
Loading amount
50 µg
Gel Running Conditions
Reduced Denaturing (4-12%)
Sample
Mouse Tissue lysate - whole (heart)
Specification
heart
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

提交于 Nov 13 2014

Application
Immunohistochemistry (Frozen sections)
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 22°C
Sample
Mouse Tissue sections (Brain)
Specification
Brain
Permeabilization
No
Fixative
Acetone
Username

Abcam user community

Verified customer

提交于 Oct 31 2014

Application
Immunohistochemistry (Frozen sections)
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 22°C
Sample
Rat Tissue sections (Muscle)
Specification
Muscle
Permeabilization
No
Fixative
Acetone
Username

Abcam user community

Verified customer

提交于 Oct 31 2014

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Sample
Human Cell (cardiomyocytes)
Specification
cardiomyocytes
Permeabilization
No
Fixative
Acetone
Username

Abcam user community

Verified customer

提交于 Oct 31 2014

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Sample
Mouse Cell (cardiomyocytes)
Specification
cardiomyocytes
Permeabilization
No
Fixative
Acetone
Username

Abcam user community

Verified customer

提交于 Oct 31 2014

Application
Western blot
Loading amount
50 µg
Gel Running Conditions
Reduced Denaturing (4-12%)
Sample
Human Cell lysate - whole cell (HeLa)
Specification
HeLa
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

提交于 Oct 06 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - nuclear (HeLa)
Loading amount
20 µg
Specification
HeLa
Gel Running Conditions
Reduced Denaturing (4-12% Bis-Tris)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 4°C
Username

Abcam user community

Verified customer

提交于 Mar 18 2013

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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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