Overview

  • Product name
  • Description
    Goat polyclonal to SNAIL
  • Host species
    Goat
  • Tested applications
    Suitable for: WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide:

    C-RKPSDPNRKPNY

    , corresponding to N terminal amino acids 8-19 of Human SNAIL

  • Positive control
    • This antibody gave a positive signal when tested against the SNAIL recombinant protein.

Properties

Applications

Our Abpromise guarantee covers the use of ab53519 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
WB Use a concentration of 0.1 - 3 µg/ml.
IHC-P Use at an assay dependent dilution.

Target

  • Function
    Involved in induction of the epithelial to mesenchymal transition (EMT), formation and maintenance of embryonic mesoderm, growth arrest, survival and cell migration. Binds to 3 E-boxes of the E-cadherin/CDH1 gene promoter and to the promoters of CLDN7 and KRT8 and, in association with histone demethylase KDM1A which it recruits to the promoters, causes a decrease in dimethylated H3K4 levels and represses transcription. Associates with EGR1 and SP1 to mediate tetradecanoyl phorbol acetate (TPA)-induced up-regulation of CDKN2B, possibly by binding to the CDKN2B promoter region 5'-TCACA-3. In addition, may also activate the CDKN2B promoter by itself.
  • Tissue specificity
    Expressed in a variety of tissues with the highest expression in kidney. Expressed in mesenchymal and epithelial cell lines.
  • Sequence similarities
    Belongs to the snail C2H2-type zinc-finger protein family.
    Contains 4 C2H2-type zinc fingers.
  • Post-translational
    modifications
    Phosphorylated by GSK3B. Once phosphorylated, it becomes a target for BTRC ubiquitination. Phosphorylation by CSNK1E, probably at Ser-104, provides the priming site for the subsequent phosphorylation by GSK3B, probably at Ser-100 and Ser-96. Phosphorylation by PAK1 may modulate its transcriptional activity by promoting increased accumulation in the nucleus. Phosphorylation at Ser-11 and Ser-92 positively regulates its functions in induction of EMT and cell survival, respectively. Phosphorylation by LATS2, upon mitotic stress, oncogenic stress or Hippo pathway activation, occurs in the nucleus and promotes nuclear retention and stabilization of total cellular protein level.
    Ubiquitinated on Lys-98, Lys-137 and Lys-146 by FBXL14 and BTRC leading to degradation. BTRC-triggered ubiquitination requires previous GSK3B-mediated SNAI1 phosphorylation. Ubiquitination induced upon interaction with NOTCH1 or TP53/p53 is mediated by MDM2.
    O-GlcNAcylation at Ser-112 is enhanced in hyperglycaemic conditions, it opposes phosphorylation by GSK3B, and stabilizes the protein.
    ADP-ribosylation by PARP1 increases protein half-life and may be involved in TGFB-induced SNAI1 up-regulation.
  • Cellular localization
    Nucleus. Cytoplasm. Once phosphorylated (probably on Ser-107, Ser-111, Ser-115 and Ser-119) it is exported from the nucleus to the cytoplasm where subsequent phosphorylation of the destruction motif and ubiquitination involving BTRC occurs.
  • Information by UniProt
  • Database links
  • Alternative names
    • dJ710H13.1 antibody
    • Protein sna antibody
    • Protein snail homolog 1 antibody
    • Protein snail homolog antibody
    • SLUGH2 antibody
    • SNA antibody
    • Sna protein antibody
    • SNAH antibody
    • SNAI antibody
    • snai1 antibody
    • SNAI1_HUMAN antibody
    • Snail 1 homolog antibody
    • Snail 1 zinc finger protein antibody
    • SNAIL antibody
    • Snail family transcriptional repressor 1 antibody
    • Snail homolog 1 (Drosophila) antibody
    • SNAIL, Drosophila, homolog of, 1 antibody
    • SNAIL1 antibody
    • Zinc finger protein SNAI1 antibody
    see all

Images

  • All lanes : Anti-SNAIL antibody (ab53519) at 1/500 dilution

    Lane 1 : SNAIL (SNAI1) Human Recombinant Protein
    Lane 2 : SLUG (SNAI2) Human Recombinant Protein

    Lysates/proteins at 0.1 µg per lane.

    Secondary
    All lanes : Rabbit polyclonal to Goat IgG - H&L - Pre-Adsorbed (HRP) (ab65486) at 1/3000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Observed band size: 68 kDa
    why is the actual band size different from the predicted?


    Exposure time: 12 minutes


    Ab53519 recognizes the tagged recombinant SNAIL protein which has an expected molecular weight of 68 kDa
  • Immunohistochemical analysis of paraformaldehyde fixed mouse tissue sections, of an urethane induced lung adenocarcinoma, labelling SNAIL in the nuclei and cytoplasm. ab53519 was used at a 1/1000 dilution in 4% donkey serum for 16 hours at 4°C. Hear mediated antigen retrieval with Citrate pH6. Blocking with 4% serum for 2 hours at 25°C. Secondary antibody was a Biotinylated Anti-Goat IgG (H+L). 

    See Abreview

  • Western blot of Rat Kidney lysate (35µg protein in RIPA buffer) staining SNAIL with ab53519 at 0.1µg/ml. Primary incubation was 1 hour. Detected by chemiluminescence.

References

This product has been referenced in:
  • Zhang B  et al. Metastasis-associated protein 2 promotes the metastasis of non-small cell lung carcinoma by regulating the ERK/AKT and VEGF signaling pathways. Mol Med Rep 17:4899-4908 (2018). Read more (PubMed: 29393472) »
  • Devis L  et al. ALCAM shedding at the invasive front of the tumor is a marker of myometrial infiltration and promotes invasion in endometrioid endometrial cancer. Oncotarget 9:16648-16664 (2018). Read more (PubMed: 29682175) »
See all 44 Publications for this product

Customer reviews and Q&As

1-8 of 8 Abreviews or Q&A

Answer

Thank you for your patience while I investigated this with the lab.

They recommended not to incubate the primary overnight. They thought this may contribute tonon-specific background after overnight incubations. They recommend to incubate for 1h at room temp. This allows a higher signal/background ratio at better controlled conditions. They thought that your observations may show that the peptide gets recruited thus enhancing the nuclear staining and they have seen this before in WB with a different antibody to a different protein.

They also recommendedto tryantigen retrieval at pH9 (10mM Tris/EDTA) instead of citrate.
I hope this information helps. Please contact us with any other questions.

Read More

Answer

Thank you for your email. I apologize for the delay as I have to leave for a conference.

I have also received an answer from lab. They have suggested that the recombinant protein they have used is tagged with GST which is why the band size seen in image is 68 kDa rather than 29-30 kDa.

Looking at the provided image and details, the antibody ab53519 seems detecting the correct target however the protocol might need optimizations in case of ab63568.

I would suggest following updates for protocol
- use 5% BSA for antibody dilutions and blocking and block membrane for 1-2 hours at RT.
- Try no primary control
- Heat lysates with sample buffer for 10 minutes at 100C.

I am sure these tips will be helpful. Should you observe similar results please do not hesitate to contact me.

Read More

Answer

Thank you for contacting us.

I apologize for the delay in this reply as I was awaiting our Products Team to add the blocking peptide to our catalogue. We have added it as ab134217. We have to update the page to show the price before you can purchase it, but you can now see it online.

https://www.abcam.com/SNAIL-peptide-ab134217.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

Read More

Answer

Thank you for contacting us.

Could you provide the following details

- Cell type e.g. Human hela cells lysates
- Provide image
- Blocking conditions for both antibodies
- Dilutions of both primary antibodies
- Information about secondary antibody
- How long the antibodies were incubated?

we will then look into details why the observed band size is different than predicted.

Read More

Answer

DISCOUNT CODE: 4XXXXX
Expiration date: 28/04/2012

I am very pleased to hear you would like to accept our offer and testab53519 in WB and IHC on human samples. This code will give you: 1 free PRIMARY ANTIBODY OR VALUE OFF OF ORDER before the expiration date. To redeem this offer, please submit an Abreview forWB and IHC on human samples and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.


Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

Read More

Answer

Thank you for contacting us. We did not test the antibody for IHC but added the application to the datasheet based on a customer's feedback. Their protocol is contained in their review at the following link, and in the reference listed at the bottom of the review, PubMed ID 20870756: https://www.abcam.com/index.html?datasheet=53519&tab=abreviews&intabreviewid=20989 I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Answer

Thanks for contacting us with your question. The rabbit SNAIL antibody ab82846 was not produced with the same immunogen as ab85931. Ab82846 has been tested for cross-reactivity with recombinant SLUG in Western blot and there is no cross-reactivity (see the WB image toward the bottom of the datasheet). However, this antibody has not been tested in IHC. The WB images show that there is quite a bit of background in endogenous samples, which means there might be some non-specific binding in IHC that could be problematic. I'm sorry to hear that the goat SNAIL antibody ab53519 produced no staining in IHC. If you would like, feel free to send some details about the protocol and I'll see if there are any suggestions to improve these results. Alternatively I can issue a credit or refund for your original purchase of ab85931. I look forward to hearing from you. Please let me know if you have any questions or if there is anything that we can do for you.

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (lung adenocarcinoma induced by urethane)
Specification
lung adenocarcinoma induced by urethane
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate pH 6
Permeabilization
No
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 25°C

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Verified customer

Submitted Oct 21 2010

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