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
  • 宿主
    Mouse
  • 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
  • 常规说明

    This product was changed from ascites to tissue culture supernatant on 12th September 2018. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

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:
  • Lu J  et al. Single-cell RNA sequencing reveals metallothionein heterogeneity during hESC differentiation to definitive endoderm. Stem Cell Res 28:48-55 (2018). ICC/IF ; Human . Read more (PubMed: 29427839) »
  • Borchard S  et al. The exceptional sensitivity of brain mitochondria to copper. Toxicol In Vitro 51:11-22 (2018). Read more (PubMed: 29715505) »

See all 30 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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