Overview

  • Product name
  • Description
    Rabbit polyclonal to Syntenin
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-P, IHC-FoFr, ICC/IF, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Xenopus laevis
    Predicted to work with: Monkey, Zebrafish
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 250 to the C-terminus of Mouse Syntenin.

    Read Abcam's proprietary immunogen policy

  • Positive control
    • HEK 293 cells, human, rat and mouse heart lysate

Properties

Applications

Our Abpromise guarantee covers the use of ab19903 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. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IHC-FoFr 1/1000.
ICC/IF Use a concentration of 1 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 32 kDa (predicted molecular weight: 32 kDa).

Target

  • Function
    Seems to function as an adapter protein. In adherens junctions may function to couple syndecans to cytoskeletal proteins or signaling components. Seems to couple transcription factor SOX4 to the IL-5 receptor (IL5RA). May also play a role in vesicular trafficking. Seems to be required for the targeting of TGFA to the cell surface in the early secretory pathway.
  • Tissue specificity
    Widely expressed. Expressed in fetal kidney, liver, lung and brain. In adult highest expression in heart and placenta.
  • Sequence similarities
    Contains 2 PDZ (DHR) domains.
  • Post-translational
    modifications
    Phosphorylated on tyrosine residues.
  • Cellular localization
    Cell junction > focal adhesion. Cell junction > adherens junction. Cell membrane. Endoplasmic reticulum membrane. Nucleus. Melanosome. Cytoplasm > cytosol. Cytoplasm > cytoskeleton. Mainly membrane-associated. Localized to adherens junctions, focal adhesions and endoplasmic reticulum. Colocalized with actin stress fibers. Also found in the nucleus. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
  • Information by UniProt
  • Database links
  • Alternative names
    • MDA-9 antibody
    • MDA9 antibody
    • Melanoma differentiation-associated protein 9 antibody
    • Pro-TGF-alpha cytoplasmic domain-interacting protein 18 antibody
    • Scaffold protein Pbp1 antibody
    • SDCB1_HUMAN antibody
    • SDCBP antibody
    • ST1 antibody
    • SYCL antibody
    • Syndecan binding protein (syntenin) antibody
    • Syndecan binding protein 1 antibody
    • Syndecan-binding protein 1 antibody
    • Syntenin 1 antibody
    • Syntenin-1 antibody
    • TACIP18 antibody
    see all

Images

  • Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
    Lane 2: Syntenin knockout HAP1 whole cell lysate (20 µg)
    Lane 3: A549 whole cell lysate (20 µg)
    Lane 4: HeLa whole cell lysate (20 µg)

    Lanes 1-4: Merged signal (red and green). Green - ab19903 observed at 32 kDa. Red - loading control, ab130007, observed at 130 kDa.

    ab19903 was shown to recognize Syntenin in wild-type HAP1 cells as signal was lost at the expected MW in Syntenin knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and Syntenin knockout samples were subjected to SDS-PAGE. ab19903 and ab130007 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1 μg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • All lanes : Anti-Syntenin antibody (ab19903) at 1 µg/ml

    Lane 1 : Mouse heart lysate
    Lane 2 : Rat heart lysate
    Lane 3 : Mouse heart lysate with Mouse Syntenin peptide (ab20431) at 1 µg/ml
    Lane 4 : Rat heart lysate with Mouse Syntenin peptide (ab20431) at 1 µg/ml

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/50000 dilution

    Predicted band size: 32 kDa
    Observed band size: 32 kDa
    Additional bands at: 50 kDa. We are unsure as to the identity of these extra bands.



    ab19903 detects a band of the expected size (32kDa) in both mouse and rat heart lysate. This band is quenched completely by the addition of the immunizing peptide in the mouse lysate and is partially quenched by the addition of ab20431 in the rat heart lysate.
  • ICC/IF image of ab19903 stained human HEK 293 cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab19903, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. 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). DAPI was used to stain the cell nuclei (blue).

  • Image courtesy of Human Protein Atlas
    ab19903 staining Syntenin in human heart tissue. Paraffin-embedded tissue was cut into 4µm sections and incubated with ab19903 (1/100 dilution) for 30 minutes at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6. ab19903 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines. Further results for this antibody can be found at www.proteinatlas.org
  • IHC-FoFr image of Syntenin staining using ab19903 (1:1000) on Mouse cortex (A) and mouse paraventricular nucleus (B). The sections used came from animals perfused fixed with Paraformaldehyde 4% with 15% of a solution of saturated picric acid, in phosphate buffer 0.1M. Following postfixation in the same fixative overnight, the brains were cryoprotected in sucrose 30% overnight. Brains were then cut using a cryostat and the immunostainings were performed using the ‘free floating’ technique.

    See Abreview

References

This product has been referenced in:
  • Verweij FJ  et al. Quantifying exosome secretion from single cells reveals a modulatory role for GPCR signaling. J Cell Biol 217:1129-1142 (2018). Read more (PubMed: 29339438) »
  • Obata Y  et al. Adiponectin/T-cadherin system enhances exosome biogenesis and decreases cellular ceramides by exosomal release. JCI Insight 3:N/A (2018). Read more (PubMed: 29669945) »
See all 7 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Application
Immunocytochemistry
Sample
Rat Cultured Cells (Hippocampus)
Permeabilization
Yes - PBS with Triton 0.25%
Specification
Hippocampus
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 20°C
Fixative
Paraformaldehyde

Luis Martins

Verified customer

Submitted May 31 2017

Answer

Thank you for you reply.

Your credit note ID is XXXXX.

If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to contact our accounting department so that we may gather the correct information needed for the refund. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used.

Please refer to the credit note ID in any correspondence with our accounting department.

The credit note ID is for your reference only and does not automatically guarantee the credit.

I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further expert advice.

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Question

Dear technical support team:
This customer has purchased ab19903 (Anti-Syntenin antibody) and has conducted the IHC assay with human sample sever times. The results show weak signal, therefore, she wants to ask for your help to modify her experiment step, could you please offer any suggestion to help her?
I also attached her data in this letter and her experiment step as follow:
1) Order details:

˙Batch number (Lot number): XXXXXXX
Po: XXXXXX
˙Abcam product code: ab19903
˙Antibody storage conditions (temperature/reconstitution etc) : -20
2) Please describe the problem (high background, non-specific signal…etc).
Weak signal
3) On what material are you testing the antibody in IHC/ICC?

˙Species: human

˙What’s cell line or tissue: lung cancer tissue

4) Sample preparation:

˙Type of sample (Fresh frozen sections, perfusion fixed frozen sections, PFA/formalin fixed

˙paraffin embedded sections, cells in culture, other:____) : paraffin embedded sections

˙Sample preparation:

˙Positive control: Hearttissue

˙Negative control:

5) Fixation step

˙Yes or No : Yes

˙If yes, Fixative agent and concentration: 10% Formalin

˙Fixation time: 2 days

˙Fixation temperature : RT

6) Antigen retrieval method(including time, temperature etc.):

Antigen Retrieval AR-10 solution (BioGenex) 121℃ 10min ( autoclave )

7) Permeabilization method:

˙Did you do a permeabilization step (details please) or add permeabilizing agent in any dilution buffers?

˙Permeabilizing agent and concentration:

8) Blocking agent (eg BSA, serum…):

˙Concentration: BioGenex Power Block

˙Blocking time : 20min

˙Blocking temperature: RT

9)

˙Endogenous peroxidases blocked? 3% H2O2, 10min

˙Endogenous biotins blocked? No

10) Primary antibody (If more than one was used, describe in “additional notes”) :

˙Species: Rabbit

˙Reacts against: Mouse, Rat, Human, Xenopus laevis

˙At what dilution(s) have you tested this antibody: 1:20

˙Diluent buffer: Antibody Diluent buffer (Ventana)

˙Incubation time: 2hr

˙Incubation temperature: RT

˙What washing steps were done (which buffer, number of washes): PBST

11) Secondary antibody:

˙Species: Super Sensitive Non-Biotin Polymer HRP Detection System BioGenex

˙Reacts against which species:

˙At what dilution(s) have you tested this antibody:

˙Diluent buffer:

˙Incubation time:

˙What washing steps were done (which buffer, number of washes):

˙Fluorochrome or enzyme conjugate (eg: FITC, HRP, AP, biotin…etc):

˙Do you know whether the problems you are experiencing come from the secondary?

12) Signal amplification method (eg: ABC, LSAB, HRP polymer, TSE): HRP polymer

13) Detection method (eg: DAB, BCIP/NBT …etc): DAB

14)

˙ How many times have you run this staining? 2 times

˙Do you obtain the same results every time? Yes

˙What steps have you altered to try and optimize the use of this antibody?

Could you please help this customer to solve the problem?

Thanks for your kindly help

Best regards

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Answer

I am sorry to hear you have been experiencing problems with one of our antibodies. The quality of our products is important to us and I would like to reassure you that we investigate all customer complaints. I appreciate the time taken to submit protocol information as well as an image to us and I would like to make some suggestions to the protocol in an attempt to improve the results obtained.

-In order to check whether the antibody is working properly, I would encourage you to use it in a tissue with high levels of the protein. According to the Human Protein Atlas, this protein is highly expressed in brain, so it could serve as a good indicator of the antibodies performance.

http://www.proteinatlas.org/ENSG00000137575

-The optimal fixation process occurs between 12 to 24 hours. If the tissue is over fixed it may well be the antibody is unable to reach the epitope.

-We usually recommend optimisation of the Antigen Retrieval method, in order to adapt it to the specific antibodies and samples used. A control experiment is recommended beforehand, where slides of the same tissue section are retrieved for 5, 10, 15, 20, 25 and 30 minutes before being immunohistochemically stained to evaluate optimum antigen retrieval time for the particular antibody being used. In the following link you will find useful information for the different antigen retrieval methods:

https://www.abcam.com/index.html?pageconfig=resource&rid=11488#notes2

It is also useful trying different buffers. In our lab we tend to use tris EDTA as it often gives successful results.

-I am not aware of the blocking agent used, but in any case, this step may also be optimised if weak staining is observed. Try milder conditions (e.g., blocking for less time, decreasing the temperature, or different agents) in an attempt to improve the results.

-We also recommend incubating the primary antibody over night at 4C to get a better staining.

-Please make sure the primary antibody and the detection system used are compatible and work successfully in other assays.

In any case, all of our products are covered by our Abpromise guarantee which ensures that you can trust our products, and they should work in the tested species and applications stated on the datasheet, or we will offer a replacement, credit, or refund, if reported within 6 months of purchase.

I hope this information is helpful. Should the suggestions not improve the results, please do not hesitate to contact me again and I will try to provide further help.

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Answer

Thank you for your phone call. As we discussed I am issuing you a credit note (for refund) for ab19903. Your credit note ID is xxx. I am sorry that this antibody did not perform as stated on the datasheet. If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to contact our accounting department so that we may gather the correct information needed for the refund. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used. Our accounting department can be contacted by email at us.credits@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website. Please refer to the credit note ID in any correspondence with our accounting department. The credit note ID is for your reference only and does not automatically guarantee the credit. I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further expert advice. If you have any further questions, please just let me know.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Mouse Tissue sections (Brain)
Specification
Brain

Dr. Sophie Pezet

Verified customer

Submitted May 04 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Xenopus laevis Cell (meiotic egg extract)
Specification
meiotic egg extract
Fixative
pre-fixation in formaldehyde and Methanol fixation after extract spindown onto coverslips
Permeabilization
No
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Mar 03 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Xenopus laevis Cell lysate - other (meiotic egg extract)
Loading amount
25 µg
Specification
meiotic egg extract
Gel Running Conditions
Reduced Denaturing (10%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Mar 03 2010

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

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