Anti-Metallothionein antibody [UC1MT] (ab12228)

Reviews (6)Q&A (25)References (45)
Western blot - Anti-Metallothionein antibody [UC1MT] (ab12228)
  • Immunocytochemistry/ Immunofluorescence - Anti-Metallothionein antibody [UC1MT] (ab12228)
  • Flow Cytometry - Anti-Metallothionein antibody [UC1MT] (ab12228)
  • Western blot - Anti-Metallothionein antibody [UC1MT] (ab12228)
  • Immunocytochemistry/ Immunofluorescence - Anti-Metallothionein antibody [UC1MT] (ab12228)

Key features and details

  • Mouse monoclonal [UC1MT] to Metallothionein
  • Suitable for: ICC/IF, WB, Flow Cyt
  • Reacts with: Rabbit, Human
  • Isotype: IgG1

Overview

  • Product name

    Anti-Metallothionein antibody [UC1MT]
    See all Metallothionein primary antibodies

  • Description

    Mouse monoclonal [UC1MT] to Metallothionein

  • Host species

    Mouse

  • Tested applications

    Suitable for: ICC/IF, WB, Flow Cytmore details

  • Species reactivity

    Reacts with: Rabbit, Human

  • Immunogen

    Full length protein corresponding to Rabbit Metallothionein. Cross-linked rabbit liver Metallothionein I and II.

  • Positive control

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

  • General notes

    This product was changed from ascites to tissue culture supernatant on 22nd May 2019. 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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

Applications

The Abpromise guarantee

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
ICC/IF
Use at an assay dependent concentration.
WB (3)
Use at an assay dependent concentration. 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 at an assay dependent concentration.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
Notes
ICC/IF
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. 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 at an assay dependent concentration.

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

Target

  • Function

    Metallothioneins have a high content of cysteine residues that bind various heavy metals; these proteins are transcriptionally regulated by both heavy metals and glucocorticoids.

  • Sequence similarities

    Belongs to the metallothionein superfamily. Type 1 family.

  • Domain

    Class I metallothioneins contain 2 metal-binding domains: four divalent ions are chelated within cluster A of the alpha domain and are coordinated via cysteinyl thiolate bridges to 11 cysteine ligands. Cluster B, the corresponding region within the beta domain, can ligate three divalent ions to 9 cysteines.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • CES 1 antibody
    • CES1 antibody
    • Metallothionein 1A antibody
    • Metallothionein 1S antibody
    • Metallothionein 2 antibody
    • Metallothionein 2A antibody
    • Metallothionein IA antibody
    • Metallothionein II antibody
    • Metallothionein-1A antibody
    • Metallothionein-IA antibody
    • Metallothionein2 antibody
    • MGC32848 antibody
    • MT 1A antibody
    • MT 2 antibody
    • MT 2A antibody
    • MT IA antibody
    • MT II antibody
    • MT-1A antibody
    • MT-IA antibody
    • MT1 antibody
    • MT1A antibody
    • MT1A_HUMAN antibody
    • MT1S antibody
    • MT2 antibody
    • MT2A antibody
    • MTC antibody
    see all

Images

  • Western blot - Anti-Metallothionein antibody [UC1MT] (ab12228)
    Western blot - Anti-Metallothionein antibody [UC1MT] (ab12228)
    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


    This image was generated using the ascites version of the product.

  • Immunocytochemistry/ Immunofluorescence - Anti-Metallothionein antibody [UC1MT] (ab12228)
    Immunocytochemistry/ Immunofluorescence - Anti-Metallothionein antibody [UC1MT] (ab12228)

    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.

    This image was generated using the ascites version of the product.

  • Flow Cytometry - Anti-Metallothionein antibody [UC1MT] (ab12228)
    Flow Cytometry - Anti-Metallothionein antibody [UC1MT] (ab12228)

    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.

    This image was generated using the ascites version of the product.

  • Western blot - Anti-Metallothionein antibody [UC1MT] (ab12228)
    Western blot - Anti-Metallothionein antibody [UC1MT] (ab12228)
    Anti-Metallothionein antibody [UC1MT] (ab12228) at 1/1000 dilution + Rabbit liver lysates

    Predicted band size: 6 kDa



    This image was generated using the ascites version of the product.

  • Immunocytochemistry/ Immunofluorescence - Anti-Metallothionein antibody [UC1MT] (ab12228)
    Immunocytochemistry/ Immunofluorescence - Anti-Metallothionein antibody [UC1MT] (ab12228)

    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.

    This image was generated using the ascites version of the product.

References (45)

Publishing research using ab12228? Please let us know so that we can cite the reference in this datasheet.

ab12228 has been referenced in 45 publications.

  • Al-Dalahmah O  et al. Single-nucleus RNA-seq identifies Huntington disease astrocyte states. Acta Neuropathol Commun 8:19 (2020). PubMed: 32070434
  • O'Doherty C  et al. Characterisation and proteomic profiling of continuously exposed Cu-resistant variants of the Caco-2 cell line. Toxicol In Vitro 65:104773 (2020). PubMed: 31981602
  • Wang B  et al. D609 protects retinal pigmented epithelium as a potential therapy for age-related macular degeneration. Signal Transduct Target Ther 5:20 (2020). PubMed: 32296021
  • Zhao Z  et al. MT2A Promotes Oxaliplatin Resistance in Colorectal Cancer Cells. Cell Biochem Biophys 78:475-482 (2020). PubMed: 32638210
  • Wu R  et al. Zn(II)-curcumin solid dispersion impairs hepatocellular carcinoma growth and enhances chemotherapy by modulating gut microbiota-mediated zinc homeostasis. Pharmacol Res 150:104454 (2019). PubMed: 31526871
View all Publications for this product

Customer reviews and Q&As

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1-10 of 31 Abreviews or Q&A

IHC - Wholemount abreview for Anti-Metallothionein antibody [UC1MT]

 Excellent
Abreviews
abreview image
Application
IHC - Wholemount
Sample
Mouse Tissue (Cornea)
Specification
Cornea
Read More

Abcam user community

Verified customer

Submitted Jan 12 2020

Western blot abreview for Anti-Metallothionein antibody [UC1MT]

 Good
Abreviews
abreview image
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
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Abcam user community

Verified customer

Submitted Jan 14 2015

Immunohistochemistry (Frozen sections) abreview for Anti-Metallothionein antibody [UC1MT]

 Good
Abreviews
abreview image
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
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Abcam user community

Verified customer

Submitted Oct 27 2014

Question


Inquiry: I was interested in looking at the western blot protocol for ab12228. However, the link leads to a page with an error message.

Read More
Answer

Thank you for contacting us.

The reason it comes as an error message is because there is no specific protocol for this antibody. The link to the general protocol is below:

https://www.abcam.com/index.html?pageconfig=resource&rid=11375


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Question

I was wondering if I could get a sample of two antibodies to test this method once before I purchase a large amount of antibody. The two antibodies for which I would like samples are as follows: Anti-Metallothionein antibody [UC1MT] (ab12228) Anti-p75 NGF Receptor antibody [NGFR5] (ab3125).

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Answer

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 datasheets. (Human, ferret, and monkey are listed on the ab3125 datasheet and human, mouse, rat and several others on the ab12228 datasheet).

Note that metallothionein is cytoplasmic, so you will not be able to sort live cells. Antibody ab3125 anti-p75 NGF Receptor, on the other hand, recognizes an extracellular epitope, so you should be able to sort live cells.

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Question

Inquiry: Hi there looking to enquire with regards to antibody MT1 ab12228. We wonder if you have a small aliquot of this that we can test for western blotting before we commit to buying in future. We have heard other groups having difficulty in getting this antibody to work hence reason for wanting to test it first. huge thanks.

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Answer

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 guarantees that the product will work in the applications and species specified on the datasheet, or we will offer a replacement, credit, or refund within 6 months of purchase.

If the product is to be used in an untested species or application, you may be eligible for our AbTrial program if the antibody has not yet been purchased. Please contact our Scientific Support team by replying to this email prior to purchase for more information. Or you can find more information about this program at the following link:

www.abcam.com/abtrial

Otherwise, we like to encourage all of our customers to submit an Abreview via the online product datasheet. We always appreciate customer feedback, whether positive or negative, and we make all product information available to researchers. Plus, each Abreview earns Abpoints that can be used for discounts on future purchases or rewards such as Amazon.com gift certificates.

To find out more about our Abreview system, please see the following link:

https://www.abcam.com/abreviews

I hope this information is helpful. Please do not hesitate to contact us again with any other questions.

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Question

Hi,



Thanks for the mail. But i cannot find the attached questionaires. Can you re-send it to me? Many thanks,

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Answer

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 understanding and co-operation in this matter. I look forward to hearing from you and hope to solve this problem as soon as possible.

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Question

Thanks so much for helping replace another MT (ab36882) antibody for our experiments. I recently have tried this replaced antibody to do some ICC (Immunocytochemistry) study; however, the result doesn't look very well. The signal is very very week (almost dark). I've tried two different concentration (1:500 and 1:1000) of this primary antibody on mouse Astroglia cells, but the data still doesn't look good. The detailed protocol please see below.

1. Cells were fixed with 4% PFA (15 min), then washed with PBS X 2 times

2. Blocking with 10% goat serum in PBST for 1 hr

3. Incubation primary Ab (ab12228 - 1:500 and 1:1000) in PBS-Tween (0.3%), overnight at 4 degrees.

4. Then wash with PBS x2, incubate with secondary Ab (Dylight-Green)- goat anti mouse 2nd Ab (diltuion 1:2000) for 3 hr.

The image still look very bad. Almost cannot see any signal. I'm wondering if we can request to replace another vial or another alternative one? If possible, please let me know. I can also send the image to you if necessary.Many thanks,

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Answer

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 attached questionnaire and provide images?

Thank you very much for your cooperation. I will look forward to hearing from you soon.

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Question

Dear xxxxx,
Thanks for your answer.
Following your recommendation, I will try to use a 12% bis tris gel with a MES running buffer, load 30µg ( I already boiled the protein). I also will load an extract of Rabbit liver containing MT2 and other isoform as a positive control.
I didn't use Cd cl2 to treat my cells, I just add a treatment who is supposed to increase the production of MT.
I let you know as soon as I have the results.
Regards,

Read More
Answer

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 the bottom).
I look forward to hearing how you get on. Until then, I hope you have a nice weekend.

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Question

Could I just clarify one more thing. In the image supplied, there is a ladder on the left and on the right. The one on the left is measuring 4.6 kDa above the 10 kDa marker on the right hand side. Do you know why this is happening?
I don't know, it's happen sometimes, but I think it's not really link to my problem...
What samples have been loaded in each well? Which cell types?
hskMDC
Are you also using milk blocking with your alpha-tubulin staining?
yes
I look forward to receiving your reply.

Read More
Answer

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 the protein ladders are detecting the expected region and that the protein has been transfered as expected.
What I would suggest doing in order to make sure the protein is not just simply moving off the gel is to run a higher % gel. We have recommended using a 4-15% gel but you could just make a 15 or 20 % gel which should make the resolution at the lower scale better as well. Just make sure not to run the protein off the gel. If you would prefer, you could always perform a dot blot before to see if there is any positive signal in your sample at all.
The only other thing I noticed was that you are currently using your secondary antibody at a dilution of 1/10,000 but the manufacturer recommends a starting point of 1/2000. It may be worthwhile to increase the concentration of the secondary to this, just to make sure that the signal is as strong as it could be. I would also increase the amount of protein loaded to 30 ug/well and perform a boiling step for 5 minutes following addition of the loading buffer.
Am I correct in thinking that you used CdCl2 with your cells prior to performing the Western blot? This can help in removing the zinc from the MT, mentioned in the following reference:
http://www.ncbi.nlm.nih.gov/pubmed/1779092
Do you have any evidence to suggest your cells should be expressing significant amounts of metallothionein? Using the Gene Expression Atlas I have not found reference to skMDC showing expression and from the following study it seems that skeletal muscle shows quite low levels when not put under stress (exercise):
http://ep.physoc.org/content/90/4/477.full
Do you have access to any other positive controls, such as liver tissue?
I look forward to receiving your reply.

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1-10 of 31 Abreviews or Q&A

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