Overview

  • Product name

    Anti-SOX6 antibody - ChIP Grade
    See all SOX6 primary antibodies
  • Description

    Rabbit polyclonal to SOX6 - ChIP Grade
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, IP, WB, IHC - Wholemount, ChIP, IHC-FoFr, CHIPseqmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 800 to the C-terminus of Mouse SOX6.

    Read Abcam's proprietary immunogen policy (Peptide available as ab30530.)

  • Positive control

    • IHC (PFA-fixed): on perfusion fixed wildtype P18 mouse dentate gyrus.

Properties

Applications

Our Abpromise guarantee covers the use of ab30455 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 1/500 - 1/1000. Antigen retrieval is not essential but may optimise staining.
IP Use at an assay dependent concentration.
WB Use at an assay dependent concentration.
IHC - Wholemount Use at an assay dependent concentration.
ChIP Use at an assay dependent concentration. PubMed: 21985497
IHC-FoFr 1/300.
CHIPseq Use at an assay dependent concentration. PubMed: 21985497

Target

  • Function

    Transcriptional activator. Binds specifically to the DNA sequence 5'-AACAAT-3'. Plays a key role in several developmental processes, including neurogenesis and skeleton formation.
  • Tissue specificity

    Expressed in a wide variety of tissues, most abundantly in skeletal muscle.
  • Sequence similarities

    Contains 1 HMG box DNA-binding domain.
  • Post-translational
    modifications

    Sumoylation inhibits the transcriptional activity.
  • Cellular localization

    Nucleus.
  • Information by UniProt
  • Database links

  • Alternative names

    • HSSOX 6 antibody
    • HSSOX6 antibody
    • SOX 6 antibody
    • sox6 antibody
    • SOX6_HUMAN antibody
    • SOXD antibody
    • SRY (sex determining region Y) box 6 antibody
    • SRY box 6 antibody
    • SRY box containing gene 6 antibody
    • Transcription factor SOX 6 antibody
    • Transcription factor Sox-6 antibody
    • Transcription factor SOX6 antibody
    see all

Images

  • IHC (PFA-fixed) image of SOX6 (ab30455) on sections of P18 mouse dentate gyrus. The mice were perfusion fixed with 4% PFA and then post-fixed in 4% PFA for a few hours. The blocks were prepared in agarose/sucrose and cut using a vibratome (50 µm). The sections were permeabilized with 1% Triton X-100 for 30 mins and blocked with 10% FBS for 2 hours at room temperature, then incubated overnight at 4°C with the primary antibody. The primary antibody was used at a 1/100 dilution and detected with donkey anti-rabbit 488 secondary at 1/1000 dilution.

  • ab30455 staining Sox6 in post-natal mouse brain sections. PFA-fixed post-natal mouse brain sections were incubated for 30 minutes in blocking solution followed by an overnight incubation with ab30455 (1/500) at 4°C. No staining was observed in sections obtained from Sox6 knockout mice.
  • IHC-FoFr image of Sox6 staining in Rat hippocampus sections using ab30455.The sections used came from animals perfused fixed with Paraformaldehyde 4%, in phosphate buffer 0.2M. Following postfixation in the same fixative overnight, the brains were cryoprotected in sucrose 30% overnight. immunostainings were performed using the ‘free floating’ technique.

    See Abreview

References

This product has been referenced in:

  • Simmons SC  et al. Determination of circuit-specific morphological adaptations in ventral tegmental area dopamine neurons by chronic morphine. Mol Brain 12:10 (2019). Read more (PubMed: 30736837) »
  • Takimoto A  et al. Differential transactivation of the upstream aggrecan enhancer regulated by PAX1/9 depends on SOX9-driven transactivation. Sci Rep 9:4605 (2019). Read more (PubMed: 30872687) »
See all 29 Publications for this product

Customer reviews and Q&As

1-6 of 6 Q&A

Answer

Thank you very much for dealing with this customer. I totally understand your concerns.

Please refund the money to customer. The credit note I have created is 22966.

Many thanks!

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Answer

Thank you for contacting us.
The current lots of this antibody are indeed 1mg/ml. Please ask for lot GR78156-2 when ordering.
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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Question
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 1049271.

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.

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Answer

Thank you for your email.

We unfortunately haveno plan, forproduction of new batch until the current stock sold out. This may happen later this year or early next year. We however are OK to send a new vial of same lot.

Let me know about your preferences.

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Question

Product code: 30455
Lot number: GR 78156-1
Inquiry: 1)Antibody storage conditions (temperature/reconstitution etc): -20°C 2)Description of the problem: no band 3)Sample: mouse skeletal muscle cells (satellite cells) 4)Sample preparation: RIPA buffer/protein inhibitors cocktail 7x/PMSF 1:100/ 5 min at 95°C 5)Amount of protein loaded: 30 ug 6)Electrophoresis/Gel conditions: 10% acrylamide gel 7)Transfer and blocking conditions: 20%methanol buffer/ 1h at 300 mA// blocked in 5% Milk (TBST1x) for 1h at RT 8)Primary Antibody: ab30455 (1:400) in 5% Milk (TBST1x) over night at 4°C 9)Secondary Antibody: anti-rabbit HRP (1:10000) for 30 min at RT 10)Detection method: ECL 11)Positive and negative controls used: I’ve compared two different stocks of the same antibody (ab30455, lot GR78156-1) on two different WB membranes with the same samples and the same amount of total proteins. I don't have the old lot number. 12)How many times have you tried the Western?: 3 times 13)Have you run a "No Primary" control?: no 14)Do you obtain the same results every time?: yes e.g. are the background bands always in the same place?: no 15)What steps have you altered?: I tried with a double concentration of the bad stock, but obtained no bands. I also tried the primary ab 1:400 in 3% BSA instead of milk. In this case many aspecific bands (over 20) appeared. Additional Notes: I attached the pictures of two different WB membranes: with the “good” aliquot of ab30455 and with the “bad” one. I also attached the WB for beta-tubulin (for the “bad” one) to normalize the total amount of proteins. The four samples consist in (from left to right): -wt satellite cells (1 day in differentiation medium) -myod-/- satellite cells (1 day in DM) -wt satellite cells (8 days in DM) -myod-/- satellite cells (8 days in DM) I can't load the image, I will send it by email

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Answer

Thank you very much for contacting us. I am very sorry to hear to hear that you are having trouble using this antibody.

We haven't received many complaints for this antibody so I am surprised to hear that this antibody completely failed. I would like to ask few questions which will help me to understand the problem better;

- Could you provide the probable date when the previous batch was purchased? Is it last month, if yes then that batch and the current batch is same.

Please note we unfortunately have same lot in stock. So would you like to try a different vial, it may well be that the vialyou have receivedis faulty.

I will look forward to hearing from you soon.

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Question

LOT NUMBER 958128 ORDER NUMBER 2011/4563 DESCRIPTION OF THE PROBLEM this antibody gives me lot of background. My collaborators reordered the same reference (unfortunately, the same lot). I tested it also and got the same absence of specific staining. SAMPLE E10.5 mouse embryo section PRIMARY ANTIBODY Anti Sox6 abcam ab30455 (polyclonal from rabbit): I tried several dilutions, inlcuding the recommended one 1/400, diluted in the blocking solution and incubated in a humidified chamber over night at 4°C. I washed the next day 3 times in PBS or PBS-Tween20 0.1% at least 10 minutes, 3 times DETECTION METHOD IF confocal microscopy POSITIVE AND NEGATIVE CONTROLS USED I am working with embryo sections. I know that Sox6 is expressed on my samples because I did IF with another antibody and got very nice signal as expected in the ventral part of the neural tube. Unfortunately, I do not have this antibody anymore and I had to find another one... ANTIBODY STORAGE CONDITIONS the antibody is stored at 4°C and was shipped already as liquid FIXATION OF SAMPLE 4%PFA 60 minutes 4°C or 0.2% PFA over night 4°C, embedded in gelatin/sucrose and cryosectionned ANTIGEN RETRIEVAL I tried without or with the antigen retrieval kit from Dako PERMEABILIZATION STEP I tried with or without permeabilization 20' in PBS-Triton X100 0.05% BLOCKING CONDITIONS I tried different blocking solution: FCS 5% in PBS Tween 20 0.1% or BSA 3% - goat serum 1% in PBS Tween 20 0.1% both 20' at least at RT SECONDARY ANTIBODY I used different Alexa from invitrogen coupled with 488 or 546 fluorochromes and made in different species. The background is not due to the secondary antibodies since I have tried several ones and I am using these secondary antibodies without any problems HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 10 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? I tried to change the fixation conditions, the blocking conditions, the washing conditions, the dilution conditions and also I tested two samples from the same lot ADDITIONAL NOTES This abcam antibody has been succesfully used by others (An CI, Dong Y, Hagiwara N., BMC Dev Biol. 2011 Oct 1011:59), that is why I ordered it...

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Answer

Thank you for contacting us. I am sorry to hear that ab30455 is producing high background in your experiment. It would be very helpful for our quality investigation if you could please provide an image of your staining. You may be able to reduce the background staining by blocking longer (up to overnight at 4C) and using 10% normal serum from the host species of your secondary antibody as the blocking agent. If your signal is strong, it may also help to reduce the primary antibody incubation time to 1-4 hours at room temperature. If you are unable to improve your results, please let me know and I would be happy to offer you a free of charge replacement with one of our other SOX6 antibodies, such as ab66316 or a credit or refund. Let me know how you would like to proceed and I will gladly assist you further.

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