Anti-Metallothionein antibody [UC1MT] (ab12228)

Overview

  • Product nameAnti-Metallothionein antibody [UC1MT]See all Metallothionein primary antibodies ...
  • Description
    Mouse monoclonal [UC1MT] to Metallothionein
  • SpecificityCross reacts with MT1 and MT2.
  • Tested applicationsIHC-Fr, ICC/IF, WB, Flow Cyt, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Rabbit, Horse, Dog, Human, Mummichug fish
  • Immunogen

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

  • Positive control
    • HeLa cell lysate treated with 100uM CdCl2 Rehydrated rabbit liver MTI/MTII

Properties

Applications

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.

Application Abreviews Notes
IHC-Fr Use at an assay dependent dilution.
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.
IHC-P Use at an assay dependent dilution.

Target

Anti-Metallothionein antibody [UC1MT] images

  • 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 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

  • 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, 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.

  • 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.

  • 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.

  • ab12228 staining human uterus tissue at 10 ug/ml using IHC-P.
  • 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

References for Anti-Metallothionein antibody [UC1MT] (ab12228)

This product has been referenced in:
  • Dai S  et al. Quantification of metallothionein on the liver and kidney of rats by subchronic lead and cadmium in combination. Environ Toxicol Pharmacol 36:1207-16 (2013). IHC-P ; Rat . Read more (PubMed: 24184865) »
  • Hoelscher M  et al. SELDI-TOF analysis of glioblastoma cyst fluid is an approach for assessing cellular protein expression. Neurol Res 35:993-1001 (2013). IHC-P ; Human . Read more (PubMed: 24225180) »

See all 12 Publications for this product

Product Wall

We do not have any free samples or small samples of any of our antibodies but both ab12228 and ab3125 will be covered by our guarantee for one year if you plan on using them for flow cytometry with samples that are one of the species listed on the data...

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Thank you for contacting us. Because we carry over 90,000 products, it isn't feasible for us to keep small sample sizes of our products.

We are happy to reassure our customers that all of our products are covered by our Abpromise, which guar...

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Thank you for your new contact.

My colleague is out of office today. I have attached the Western blot Questionnaire to this e-mail and would be grateful if you could complete it and send it back to us.

Thank you for your understandi...

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Thank you for your email. I am sorry to hear that you have been experiencing problems with this antibody.

I have read the details you have kindly provided and will be very happy to investigate the antibody further.

- Please fill the ...

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Thank you for getting back to me.
I think that sounds like a good idea. I would say that trying a 15% or 20% gel would probably perform better. Otherwise, make sure that the protein is not run off the gel (stop it well before the dye front reached...

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Thank you for getting back to me with this information and sorry for the delay in my reply.
The reason I asked about the ladder is because the Metallothionein protein is very small and it is therefore important to make sure that the lower end of t...

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Thank you for your reply.
I'd like to clarify a few details if you wouldn't mind in order to understand the problems encountered with the Anti-Metallothionein antibody [UC1MT] (ab12228) more fully.
1. You mentioned that with 10 ug loaded ther...

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Thank you for contacting us and reporting the problems you have been experiencing with the Anti-Metallothionein antibody [UC1MT] (ab12228).
I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints v...

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Thank you for enquiry regarding ab12228.
I apologize for the long delay in my answer. Indeed, there is some confusion about utility of the addition to the thimerosal to the Western blot.
To summarize, you do not necessarily need to add it. Read More

Thank you for your inquiry.

Unfortunately, we are not able to release the immunogen sequence for this particular antibody as it is proprietary information.

We aim to provide as much information as possible to our customers, so I am so...

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