Overview

  • Product name
  • Description
    Rabbit polyclonal to TMEM16A
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-P, ICC/IF, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Positive control
    • GIST

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer
    pH: 7.40
    Preservative: 0.1% Sodium azide
    Constituents: 1% BSA, 0.0268% PBS
  • Concentration information loading...
  • Purity
    Protein G purified
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab53212 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-P 1/100.
ICC/IF Use a concentration of 2 µg/ml. PubMed: 19819874
IHC-Fr Use at an assay dependent concentration. PubMed: 21084687

Target

  • Function
    Acts as a calcium-activated chloride channel. Required for normal tracheal development.
  • Tissue specificity
    Broadly expressed with higher levels in liver and skeletal muscle.
  • Sequence similarities
    Belongs to the anoctamin family.
  • Domain
    The region spanning the fifth and sixth transmembrane domains probably forms the pore-forming region.
  • Cellular localization
    Cell membrane. Cytoplasm.
  • Information by UniProt
  • Database links
  • Alternative names
    • ANO 1 antibody
    • ANO1 antibody
    • ANO1_HUMAN antibody
    • Anoctamin 1 antibody
    • Anoctamin 1 calcium activated chloride channel antibody
    • Anoctamin-1 antibody
    • Anoctamin1 antibody
    • Ca2+ activated Cl- channel antibody
    • Calcium Activated Chloride Channel antibody
    • Discovered on gastrointestinal stromal tumors protein 1 antibody
    • DOG 1 antibody
    • DOG1 antibody
    • FLJ10261 antibody
    • Membrane protein antibody
    • Oral cancer overexpressed 2 antibody
    • Oral cancer overexpressed protein 2 antibody
    • ORAOV 2 antibody
    • ORAOV2 antibody
    • TAOS 2 antibody
    • TAOS2 antibody
    • TMEM 16A antibody
    • TMEM16A antibody
    • Transmembrane protein 16A (eight membrane spanning domains) antibody
    • Transmembrane protein 16A antibody
    • Tumor amplified and overexpressed sequence 2 antibody
    • Tumor-amplified and overexpressed sequence 2 antibody
    see all

Images

  • Immunocytochemistry/ Immunofluorescence analysis of Wt1cre-YFP mouse interstitial cell of Cajal labelling TMEM16A with ab53212 at 1/20 dilution. Cells from the muscular layer, especially the circular one, show TMEM16A (ANO1 - red) immunoreactivity. This immunostaining becomes strong by E16.5. Colocalization of YFP with ANO1 is observed in many cells of both, the circular and the longitudinal layer.

  • ab53212 (2µg/ml) staining TMEM16A in human liver (left panel) using an automated system (DAKO Autostainer Plus). Using this protocol there is strong staining of hepatocyte cell membrane.
    Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
  • Analysis of TMEM16A expression was performed in frozen liver sections (10 µm) from normal and 7-day bile duct-ligated rats and mice.
    Sections were defrosted at room temperature and fixed in 4% paraformaldehyde (1× PBS), permeabilized in PBST (1× PBS with 0.2% Triton), and blocked in 4% BSA (in PBST) for 1 hour at room temperature. Samples were then incubated with ab53212 at a 1/100 dilution, diluted in 1% BSA. Sections were incubated overnight at 4 °C and washed three times for 10 minutes each with 1× PBST at room temperature. Sections were incubated with Dylight 488 conjugated donkey anti-rabbit secondary at a 1/600 dilution for 2 hours at room temperature protected from light. Following incubation, the slides were washed in PBST at room temperature and coverslipped with Antifade gold and DAPI. Images were visualized using an Olympus IX-71 confocal microscope.

    Scale bar, 10 µm.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human gastrointestinal stromal tumor labeling TMEM16A with ab53212.

  • ICC/IF image of ab53212 stained HepG2 cells. The cells were 4% formaldehyde 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 (ab53212, 1µg/ml) 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.

References

This product has been referenced in:
  • Wang P  et al. Inflammatory mediators mediate airway smooth muscle contraction through a G protein-coupled receptor-transmembrane protein 16A-voltage-dependent Ca2+ channel axis and contribute to bronchial hyperresponsiveness in asthma. J Allergy Clin Immunol 141:1259-1268.e11 (2018). WB, ICC/IF . Read more (PubMed: 28754608) »
  • Song Y  et al. Inhibition of ANO1/TMEM16A induces apoptosis in human prostate carcinoma cells by activating TNF-a signaling. Cell Death Dis 9:703 (2018). Read more (PubMed: 29899325) »
See all 36 Publications for this product

Customer reviews and Q&As

1-10 of 12 Abreviews or Q&A

Abcam has not validated the combination of species/application used in this Abreview.
Application
IHC - Wholemount
Sample
Guinea pig Tissue (intestine myenteric plexus)
Specification
intestine myenteric plexus

Abcam user community

Verified customer

Submitted Nov 29 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (skin)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Citrate pH6
Permeabilization
No
Specification
skin
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Jul 11 2016

Answer

Thank you for your reply and for kindly providing the word document. This has made conducting the alignments much more straight forward.

I am sorry to confirm that as far as we are aware,none of our TMEM16A antibodieshas been tested with samples from Zebrafish. All tested species covered by the 6 month guarantee are stated on our datasheets, and these are updated as soon as any new information is brought to our attention.

I have obtained the immungoen sequences for all our TMEM16A antibodies and checked the alignment of these with the 3 Zebrafish sequences you provided.

Regrettably, for most of these the alignments are too low for us to suggest they may detect in Zebrafish. However, there is one with reasonable alignment that you may like to consider trying. I can recmomend to review the online datasheet for further information.

ab84115
https://www.abcam.com/index.html?datasheet=84115 (or use the following: https://www.abcam.com/index.html?datasheet=84115).

Immunogen: ################

Alignment with Zebrafish sequences provided:
Sequence 1: 68%
Sequence 2: 68%
Sequence 3: 72%

We recommend alignment should be over 85% to predict that an antibody will detect. It isstill possible that with 68-72% alignment therecould be some crossreactivity. However, even when alignmentis very highwe cannot guarantee this without further testing.


If you would like to test the ab84115 in Zebrafish, please contact me again by replying to this message prior to the purchase as you may be eligible for our testing discount program.

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. 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 me for any further advice or information.

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Question
Answer

Thank you for your call last week and for your patience while I have been in touch with the lab regarding your enquiry.

Unfortunately we are not able to provide the blocking peptide for ab53212 at this time. I am very sorry that this product is not available, but if there is anything else that we can do for you please let me know and I'll be happy to help.

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Answer

Thank you for your inquiry.

I can confirm that the IHC-Fr image was not generated by our lab but is taken from the following publication:

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3013035/?tool=pubmed

Please follow the above link to find out more about the protocol they used.

I also would like to point out that this publication used aDylight 488 conjugated donkey anti rabbit antibody that was purchased from a competitor and I can therefore not comment on the dilution that would be suitable.

But I am happy to confirm that antibodies from our catalog are also Dylight 488 conjugated donkey anti rabbit antibodies and suitable for your customers purpose. They can be used at a dilution (of the stock in the delivered vial) of 1;50 to 1:500.

ab98488 and ab96919 weresuccessfully used in a 1:200 dilution. To review the images please also follow the link to the respective datasheets:

ab98488

https://www.abcam.com/index.html?datasheet=98488 (or use the following: https://www.abcam.com/index.html?datasheet=98488).

ab96919

https://www.abcam.com/index.html?datasheet=96919 (or use the following: https://www.abcam.com/index.html?datasheet=96919).

I hope this information is helpful for your customer and wish them good luck with their research.

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Answer

Thank you for contacting us.
This image is from publication; Dutta AK et al, J Biol Chem. 2011 Jan 7;286(1):766-76. Epub 2010 Nov 1, Fig 1. DOI 10.1074/jbc.M110.164970. Customer can ask this to Author of the paper.
We also have DAPI mounting media in catalogue. The catalogue numbers are ab104139, ab10140
https://www.abcam.com/Fluoroshield-Mounting-Medium-With-DAPI-20ml-ab104139.html
https://www.abcam.com/Fluoroshield-Mounting-Medium-With-DAPI-ab104140.html
I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.
Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

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Answer

Thanks for your reply.

To calculate the 89% score, did you use the canis lupus sequence that I sent or a different dog sequence? I ran the BLAST again and the Identity score came up as 94%, so I'm not sure if we're looking at the same canine sequence.

Another option would be ab84115, which gave an Identity score of 98% with the canis lupus sequence-

https://www.abcam.com/TMEM16A-antibody-ab84115.html

If you would like to try either of these antibodies in canine samples, please let me know because we do have a testing discount program that you will be eligible for. In this program, you would purchase the TMEM16A antibody, try it in canine samples, submit an Abreview with your data, and in exchange we will give you one free primary antibody on a future order. Please let me know if this is something you're interested in and I'll send more information.

Please let me know if you have any questions or if there is anything else that I can do for you.

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Answer

Thank you for your enquiry. I am pleased to provide the following suggested positive controls for these antibodies. I have mainly chosen tissue that has been tested succesfully and shown in an image on the datasheets: 1.Anti-ErbB 2 antibody (ab2428) Human breast cancer 2.Anti-Progesterone Receptor antibody [SP2] (ab16661) Human breast carcinoma 3.Anti-Estrogen Receptor antibody [SP1] (ab16660) Human breast carcinoma 4.Anti-Myeloperoxidase antibody (ab45977) Human spleen tissue 5.Anti-TdT antibody (ab85148) Human testis 6.Anti-TMEM16A antibody (ab53212) Human liver 7.Anti-NSE antibody [EPR3377] (ab79757) Human brain tissue 8.Anti-CD23 antibody [EPR3617(IHC)] (ab92495) Human tonsil 9.Anti-CD10 antibody [56C6] (ab951) Lymph node tissue or human breast microepithelium 10.Anti-Melanoma antibody [HMB45 + DT101 + BC199] (ab732) Murine melanoma metastasis tissue 11.Anti-CD79a antibody [HM47/A9] (ab3121) Human Lymphoid tissue, tonsil 12.Anti-CD45RO antibody [UCHL-1] (ab86080) Human Tonsil, Thymus 13.Anti-CD45 antibody (ab10559) Human Tonsil tissue 14.Anti-CD30 antibody [CON6D/ C2] - BSA and Azide free (ab76518) Human Hodgekins lymphoma tissue 15.Anti-CD15 antibody [(Workshop IV) (BRA-4F1) our designation 238)] (ab115993) Human Hodgekins lymphoma tissue Human spleen I am providing a list ofslides that we sell which I hope will be helpful to you. I can recommend to review the online datasheet for more details. Human brain: ab4305Brain (human): hippocampus normal tissue slides https://www.abcam.com/index.html?datasheet=4305 (or use the following: https://www.abcam.com/index.html?datasheet=4305). Human lymph node: ab4350Lymph node (human) normal tissue slides IHC-P, ISH https://www.abcam.com/index.html?datasheet=4350 (or use the following: https://www.abcam.com/index.html?datasheet=4350). Human testis: ab4373Testis (human) normal tissue slides IHC-P, ISH https://www.abcam.com/index.html?datasheet=4373 (or use the following: https://www.abcam.com/index.html?datasheet=4373). Human liver: ab4348Liver (human) normal tissue slides IHC-P, ISH https://www.abcam.com/index.html?datasheet=4348 (or use the following: https://www.abcam.com/index.html?datasheet=4348). Human spleen: ab4372Spleen (human) normal tissue slides IHC-P, ISH https://www.abcam.com/index.html?datasheet=4372 (or use the following: https://www.abcam.com/index.html?datasheet=4372). I am sorry we do not have slides forthe following: Human tonsil Hodgekins lymphoma tissue Mouse melenoma tissue I hope this will be helpful to you. If you have any further questions,please do not hesitate to contact us.

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Answer

Thank you for contacting us. Unfortunately, we do not currently have a blocking peptide for ab53212 available for purchase. The immunogen for this antibody is located in the range between amino acids 100- 450 of human TMEM16A. You may be able to have a peptide synthesized for this region through a company such as 21st Century Biochemicals. We do have an antibody/ peptide pair available to TMEM16A as ab84915 and ab97423. I hope this helps, please let me know if you need any additional information.

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1-10 of 12 Abreviews or Q&A

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