Overview

  • Product name
    Anti-gamma Synuclein antibody
    See all gamma Synuclein primary antibodies
  • Description
    Rabbit polyclonal to gamma Synuclein
  • Host species
    Rabbit
  • Specificity
    The immunizing peptide was chosen to avoid cross-reactivity of the antiserum to other synucleins. Immunohistochemical and western blot analysis of human brain indicates a high level of specificity for this antiserum.
  • Tested applications
    Suitable for: ICC/IF, IP, IHC-P, IHC-Fr, WBmore details
  • Species reactivity
    Reacts with: Rat, Human
  • Immunogen

    Synthetic peptide

    EKEEVAEEAQSGGD

    corresponds to amino acids 114-127 of the human gamma synuclein protein. The conjugate was cross-linked by maleimidocaproyl-N-hydroxy succinimide (MSC) and coupled to diphtheria toxoid.

Properties

Applications

Our Abpromise guarantee covers the use of ab6169 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
IP
IHC-P
IHC-Fr
WB
  • Application notes
    ICC/IF: 1/100 - 1/150.
    IHC-P: 1/100.
    IHC-Fr: Use at an assay dependent dilution.
    WB: 1/500 - 1/1000.
    1 site ELISA: Not tested.

    Not yet tested in other applications.
    Optimal dilutions/concentrations should be determined by the end user.
  • Target

    • Function
      Plays a role in neurofilament network integrity. May be involved in modulating axonal architecture during development and in the adult. In vitro, increases the susceptibility of neurofilament-H to calcium-dependent proteases (By similarity). May also function in modulating the keratin network in skin. Activates the MAPK and Elk-1 signal transduction pathway.
    • Tissue specificity
      Highly expressed in brain, particularly in the substantia nigra. Also expressed in the corpus callosum, heart, skeletal muscle, ovary, testis, colon and spleen. Weak expression in pancreas, kidney and lung.
    • Sequence similarities
      Belongs to the synuclein family.
    • Post-translational
      modifications
      Phosphorylated. Phosphorylation by GRK5 appears to occur on residues distinct from the residue phosphorylated by other kinases.
    • Cellular localization
      Cytoplasm > perinuclear region. Cytoplasm > cytoskeleton > centrosome. Cytoplasm > cytoskeleton > spindle. Associated with centrosomes in several interphase cells. In mitotic cells, localized to the poles of the spindle.
    • Information by UniProt
    • Database links
    • Alternative names
      • BCSG1 antibody
      • Breast cancer specific gene 1 protein antibody
      • Breast cancer specific protein 1 antibody
      • Breast cancer-specific gene 1 protein antibody
      • Gamma synuclein antibody
      • Gamma-synuclein antibody
      • Persyn antibody
      • PRSN antibody
      • SNCG antibody
      • SR antibody
      • Synoretin antibody
      • Synuclein gamma antibody
      • synuclein, gamma (breast cancer-specific protein 1) antibody
      • SYUG_HUMAN antibody
      see all

    Images

    • ab6169 was tested on recombinant protein only, 100ng recombinant protein each lane. 1= alpha-synuclein, 2 =  beta-synuclein, 3 = gamma-synuclein. The primary was 1/2000, secondary was alkaline phosphatase conjugated anti-rabbit from Sigma. We did not try it on crude samples such as brain or other tissue homogenates.

      ab6169 was tested on recombinant protein only, 100ng recombinant protein each lane. 1= alpha-synuclein, 2 =  beta-synuclein, 3 = gamma-synuclein. The primary was 1/2000, secondary was alkaline phosphatase conjugated anti-rabbit from Sigma. We did not try it on crude samples such as brain or other tissue homogenates.

    • ICC/IF image of ab6169 stained MCF7 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 (ab6169, 5µg/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.

    • IHC image of ab6169 staining in human breast carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab6169, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    References

    This product has been referenced in:
    • He X  et al. Long non-coding RNA AK058003, as a precursor of miR-15a, interacts with HuR to inhibit the expression of ?-synuclein in hepatocellular carcinoma cells. Oncotarget 8:9451-9465 (2017). WB, ICC/IF, Other ; Human . Read more (PubMed: 28035067) »
    • Newman AJ  et al. A New Method for Quantitative Immunoblotting of Endogenous a-Synuclein. PLoS One 8:e81314 (2013). WB ; Human . Read more (PubMed: 24278419) »
    See all 10 Publications for this product

    Customer reviews and Q&As

    1-10 of 10 Abreviews or Q&A

    Abcam has not validated the combination of species/application used in this Abreview.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Retina)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate Buffer
    Permeabilization
    Yes - Tween-20
    Specification
    Retina
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
    Fixative
    Davidson

    Abcam user community

    Verified customer

    Submitted Sep 20 2018

    Question

    Dear technical support team:

    This customer has purchased ab6169 (Anti-gamma Synuclein antibody) and has conducted the wb several times with rat sample. The results show no band, therefore, she would like to ask for your help to modify her experiment step, could you please offer any suggestion to improve her result?

    Her experiment step as follow:



    1. Order details:


    Batch number:

    Po: 988946

    Abcam product code: ab6169

    Antibody storage conditions (temperature/reconstitution etc) : -20 Celsius




    2. Please describe the problem (high background, wrong band size, more bands, no band etc). I can’t get the correct band at the predict size.



    3. On what material are you testing the antibody in WB?

    · Species:

    · What’s cell line or tissue : rat tendon fibroblast

    · Cell extract or Nuclear extract: cell extract

    · Purified protein or Recombinant protein: purified



    3. The lysate


    How much protein was loaded: 20 uM

    What lysis buffer was used: NE lysis



    What protease inhibitors were used: no

    What loading buffer was used: 2X sample dye

    Phosphatase inhibitors :no

    Did you heat the samples: temperature and time: yes I did heat at 95 Celsius




    4. Electrophoresis/Gel conditions/ Transfer conditions


    Reducing or non reducing gel:

    Reducing agent:

    Gel percentage : 15%

    Transfer conditions: (Type of membrane, Protein transfer verified): 55 V , 2hours




    5. Blocking conditions


    Buffer: 1X PBS

    Blocking agent: milk, BSA, serum, what percentage: no fat milk 5%

    Incubation time: 30mins

    Incubation temperature: RT




    6. Primary Antibody


    Species: rabbit polyclonal

    Reacts against: Hu , Rat


    · At what dilution(s) have you tested this antibody: 1:500

    · What dilution buffer was used: 1XPBS

    · Incubation time: 2 hours

    · Incubation temperature: RT

    · What washing steps were done: 1XPBS , 15mins two time



    7. Secondary Antibody


    Species: anti rabbit

    Reacts against:

    At what dilution(s) have you tested this antibody: 1:2000

    Incubation time: 1hour

    Wash steps: 1XPBS , 15mins two time

    Fluorochrome or enzyme conjugate:

    Do you know whether the problems you are experiencing come from the secondary? No




    8. Detection method
    ECl, ECl+, other detection method: ECL +



    9. Did you apply positive and negative controls along with the samples? Please specify. No I don’t

    10. Optimization attempts

    · How many times have you tried the Western? At least 8 times

    · Have you eliminated the possibility that any background bands could be due to the secondary antibody? (Run a “No primary” control): no, I don’t think so

    · Do you obtain the same results every time e.g. are background bands always in the same place? yes

    · What steps have you altered? Protein loading and dilution of primary antibody

    ·

    Could you please help this customer to solve the problem?

    Thanks for your kindly help

    Best regards

    Read More
    Answer

    Thank you and your customer for taking the time to complete our questionnaire and contact us. I am sorry to hear your customer has had difficulty obtaining satisfactory results from this antibody.

    The details your customer has kindly provided will enable us to investigate this case for your customer and this is also helpful in our records for monitoring of quality.

    Reviewing this case, I would like to offer some suggestions to help optimise the results from ab6169:

    1.) I suggest to use positive control. Wehave positive data with MCF7 cells. I could not find any evidence that gamma Synuclein is expressed in rat tendons or fibroblasts.
    It is described as specifically expressed in the peripheral nervous system with high expression in motoneurons of the brainstem and it lso found in neurons of many other brain regions including the cerebellar cortex, thalmus, hypothalamus and CA1, CA2, CA3 and CA4 regions of the hippocampus.

    http://www.uniprot.org/uniprot/Q63544

    http://biogps.org/#goto=genereport&id=6623(choose species rat)

    2.) General suggestion would include to incubate the primary antibody over night at 4C and to use BSA as blocking agent. This would ensure a stronger signal.

    We are happy to offer this technical support. In the event that a product is not functioning in the applications cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund.

    I hope this information is helpful, thank you and your customer for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again.

    Read More

    Answer

    Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number xxxx. To check the status of the order please contact our Customer Service team and reference this number. Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know. I wish you the best of luck with your research.

    Read More

    Answer

    Thank you for contacting Abcam. At Abcam, providing quality service is just as important as providing quality products. Our Abpromise® ensures that you can trust our products to perform as stated on the datasheets, or we will offer a replacement, credit, or refund. For our records, could you please send me the protocol that you have used with ab47966. Including cell preparation, blocking solutions, times and temperature as well as antibody incubation times and any controls that you may have used. If possible, would you be able find the lot number of the previous vial or an order number from that purchase as well? The Abcam Abpromise® guarantee: • 100% Scientific and Customer Support for any product you buy from Abcam or one of our authorized distributors. • We guarantee our products work in the tested species and applications stated on the datasheet. • We will replace or refund products not performing as stated on the datasheet if reported within 6 months of purchase. • We investigate any quality concerns raised by customers to ensure our catalog contains products that perform to the highest standards. Please note these conditions to our Abpromise®: • Protocol information must be provided in order for the claim to be validated. • Antibodies tested in recombinant samples only are not guaranteed for use on endogenous samples. • “Predicted to react” information on the datasheets is provided for reference only and these species are not guaranteed. • Fast Track antibodies are covered for ELISA against the immunizing peptide only. I look forward to hearing back form you and working through this issue with you. I hope this information is helpful, but please do not hesitate to contact us with further questions.

    Read More

    Answer

    Thank you for your enquiry. The only gamma-synuclein antibody which we currently have is ab6169, which is not directed against mouse. To see if another company may have what you are looking for, I suggest that you click on the link to "The World's Antibody Gateway". The World's Antibody Gateway is a free search engine service provided by Abcam to help you to quickly find the antibodies that you are looking for. It is a free-text search engine developed by the Abcam team so that it searches the catalogs of all online antibody companies (currently 249).

    Read More

    Answer

    We tested the antibody on recombinant protein only 100ng recombinant protein each lane. The primary was 1/2000, secondary was alkaline phosphatase conjugated anti-rabbit from Sigma. We did not try it on crude samples such as brain or other tissue homogenates. I have attached the Western blot image to the datasheet so please refer to that.

    Read More

    Answer

    The antibody should work on paraffin embedded breast cancer tissue, as it works fine with paraffin embedded human brain tissue which contained abundant gamma-synuclein. However, we have not tried any non-neural tissues.

    Read More

    Answer

    For neurological tissue, the use of a scientific microwave is preferable, as it can be set to just below boiling to help minimise the risk of section dissociation. We have no experience of this particular antibody in non-neural tissue. For a general antigen retrieval and IHC protocol on formalin fixed paraffin embedded tissue, please see the links on the right hand side of the Abcam homepage.

    Read More

    Answer

    Try using this antibody at the recommended dilution of 1:1000, then adjust the concentration accordingly from the results. Make sure you perform antigen retrieval before staining (I'll send you my protocol). If doing immunofluorescence, use PBS for buffer washes and to make up reagents. If doing immunoperoxidase or phosphatase use TBS with 0.025% Triton for buffer washes (except after the application of the secondary, use TBS alone). If you're not using a kit for immunoperoxidase or phosphatase, make all reagents up in TBS with 1% BSA. (Leave the BSA out of the ABC, if you're using it, and the chromagen). Use a humidified incubation chamber to avoid slides drying out. Block in normal rabbit serum for 2 hours before adding the primary. Use at 1:10 Incubate the primary at 4 degrees centigrade over night. If using HRP, block endogenoue peroxidases with hydrogen peroxide after adition of the secondary. Likewise if using AP, but use Levamisole. If using AEC as a chromagen, use an aqueous mountant and do not dehydrate through alcohol. I hope that these tips are of use to you.

    Read More

    Question
    Answer

    If the centriole contains tubulin then this antibody will detect it.

    Read More

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