Anti-Metallothionein抗体[UC1MT] (ab12228)

概述

  • 产品名称
    Anti-Metallothionein抗体[UC1MT]
    参阅全部 Metallothionein 一抗
  • 描述
    小鼠单克隆抗体[UC1MT] to Metallothionein
  • 特异性
    Cross reacts with MT1 and MT2.
  • 经测试应用
    适用于: IHC-Fr, ICC/IF, WB, Flow Cyt, IHC-Pmore details
  • 种属反应性
    与反应: Mouse, Rat, Rabbit, Horse, Dog, Human, Mummichug fish
  • 免疫原

    Full length protein (cross linked rabbit liver MT1 and MT2).

  • 阳性对照
    • HeLa cell lysate treated with 100uM CdCl2 Rehydrated rabbit liver MTI/MTII

性能

应用

Our Abpromise guarantee covers the use of ab12228 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-Fr Use at an assay dependent concentration.
ICC/IF Use a concentration of 1 - 5 µg/ml.
WB 1/1000. Detects a band of approximately 6-20 kDa (predicted molecular weight: 6 kDa). Please note: often Western blots done on cell lysates with this antibody produce many bands; we suspect that metallothionein binds to many other proteins, thus producing these results. As the predicted MW is around 6 kDa, use 12.5-20% gel and be sure the protein is not run off the gel during electrophoresis.
Flow Cyt Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

IHC-P Use at an assay dependent concentration.

靶标

图片

  • Anti-Metallothionein antibody [UC1MT] (ab12228) + Hela cell lysate

    Secondary
    HRP-conjugated antibody.
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 6 kDa


    Exposure time : 2 minutes
  • ab12228 staining human uterus tissue at 10 ug/ml using IHC-P.

  • ICC/IF image of ab12228 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 (ab12228, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse 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.

  • Overlay histogram showing HeLA cells stained with ab12228 (red line). The cells were fixed with 100% 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 (ab12228, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. 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-Metallothionein antibody [UC1MT] (ab12228) at 1/1000 dilution + Rabbit liver lysates

    Predicted band size : 6 kDa
  • ICC/IF image of ab12228 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 (ab12228, 10µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-mouse IgG - H&L, pre-adsorbed (ab96879) 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.

  • All lanes : Anti-Metallothionein antibody [UC1MT] (ab12228)

    Lane 1 : Marker
    Lane 2 : MTI
    Lane 3 : MTII
    Lane 4 : Mummichug CdCl2


    Predicted band size : 6 kDa
  • ab12228 staining catfish kidney tissue sections by IHC-Fr.  Sections were acetone fixed and blocked with a commercial blocking agent prior to incubation with the primary antibody, diluted 1/50, for 16 hours at 4°C.  An Alexa Fluor® 488 conjugated goat anti-mouse antibody, diluted 1/1000, was used as the secondary.

    See Abreview

文献

This product has been referenced in:
  • Kadota Y  et al. Metallothioneins regulate the adipogenic differentiation of 3T3-L1 cells via the insulin signaling pathway. PLoS One 12:e0176070 (2017). ICC/IF . Read more (PubMed: 28426713) »
  • Sullivan B  et al. On the nature of the Cu-rich aggregates in brain astrocytes. Redox Biol 11:231-239 (2017). Read more (PubMed: 28012438) »

See all 27 Publications for this product

客户评价及客户问答

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Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Reduced Denaturing (15% acrylamide)
Sample
Human Cell lysate - whole cell (osteosarcoma)
Specification
osteosarcoma
Blocking step
blocking buffer Sigma as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 20°C
Username

Abcam user community

Verified customer

提交于 Jan 14 2015

Application
Immunohistochemistry (Frozen sections)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10 · Temperature: 23°C
Sample
Rat Tissue sections (Brain)
Specification
Brain
Permeabilization
Yes - Methanol
Fixative
Methanol
Username

Abcam user community

Verified customer

提交于 Oct 27 2014

Application
Western blot
Sample
Human Cell lysate - whole cell (LNCaP (prostate cancer cell line))
Loading amount
10 µg
Specification
LNCaP (prostate cancer cell line)
Gel Running Conditions
Reduced Denaturing (7% tris-acetate gel)
Blocking step
Milk as blocking agent for 35 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Abcam user community

Verified customer

提交于 Jul 23 2008

Application
Immunohistochemistry (Frozen sections)
Sample
Catfish Tissue sections (kidney)
Specification
kidney
Fixative
Acetone
Permeabilization
No
Blocking step
Image-iT as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C
Username

Dr. David Matlack

Verified customer

提交于 Jun 09 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Rat Tissue lysate - whole (choroid plexus(CP) and kidney(K))
Loading amount
30 µg
Specification
choroid plexus(CP) and kidney(K)
Gel Running Conditions
Reduced Non-Denaturing (Native) (Gel 12,5%)
Blocking step
Casein as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 20°C
Username

Abcam user community

Verified customer

提交于 Mar 06 2008

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

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