Overview

  • Product name
    Anti-SCD1 antibody [CD.E10]
    See all SCD1 primary antibodies
  • Description
    Mouse monoclonal [CD.E10] to SCD1
  • Host species
    Mouse
  • Tested applications
    Suitable for: IHC-Fr, IHC-P, ICC/IF, Flow Cyt, WB, IP, ELISAmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant full length protein (Human).

  • Positive control
    • NIH 3T3 cells
  • General notes

    SCD1 is known to undergo post-translational modifications and the sizes differ in different cell lines so the observed band size can be different than predicted band size. As positive control we recommend using SCD1 over-expressed 293 transfected cell lysates for western blot.

    This product was changed from ascites to tissue culture supernatant on 25th May 2018. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

Properties

Applications

Our Abpromise guarantee covers the use of ab19862 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-Fr Use at an assay dependent concentration.
IHC-P Use a concentration of 4 µg/ml.
ICC/IF Use at an assay dependent concentration. PubMed: 20530718
Flow Cyt Use 1µg for 106 cells.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

 

WB 1/1000. Predicted molecular weight: 41.5 kDa. PubMed: 18697866
IP Use at an assay dependent concentration.
ELISA Use at an assay dependent concentration.

Target

  • Function
    Terminal component of the liver microsomal stearyl-CoA desaturase system, that utilizes O(2) and electrons from reduced cytochrome b5 to catalyze the insertion of a double bond into a spectrum of fatty acyl-CoA substrates including palmitoyl-CoA and stearoyl-CoA.
  • Sequence similarities
    Belongs to the fatty acid desaturase family.
  • Domain
    The histidine box domains may contain the active site and/or be involved in metal ion binding.
  • Cellular localization
    Endoplasmic reticulum membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • ACOD_HUMAN antibody
    • Acyl-CoA desaturase antibody
    • Delta(9)-desaturase antibody
    • Delta-9 desaturase antibody
    • Delta-9-Desaturase antibody
    • FADS5 antibody
    • Fatty acid desaturase antibody
    • PRO0998 antibody
    • SCD 1 antibody
    • SCD antibody
    • SCD1 antibody
    • Stearoyl-CoA desaturase antibody
    see all

Images

  • Western blot using ab19862 on 293 cells. Western blot using ab19862 on 293 cells.
  • SCD was immunoprecipitated using 0.5mg Hek293 whole cell extract, 10µg of Mouse monoclonal to SCD and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, Hek293 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab19862.
    Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution.
    Band: 32kDa; SCD.
  • ab19862 staining SCD in human skin.
    Left panel: with primary antibody at 4 ug/ml. Right panel: isotype control.
    Sections were stained using an automated system (DAKO Autostainer Plus), at room temperature: sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffers EDTA pH 9.0. 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 we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
  • ICC/IF image of ab19862 stained HeLa cells. The cells were 100% methanol fixed (5 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 (ab19862, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse 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.

  • Overlay histogram showing HepG2 cells stained with ab19862 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab19862, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HepG2 cells fixed with 80% methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

References

This product has been referenced in:
  • Sanchez DJ  et al. PPAR? is dispensable for clear cell renal cell carcinoma progression. Mol Metab N/A:N/A (2018). WB ; Human . Read more (PubMed: 29866440) »
  • Xie H  et al. IRE1a RNase-dependent lipid homeostasis promotes survival in Myc-transformed cancers. J Clin Invest 128:1300-1316 (2018). Read more (PubMed: 29381485) »
See all 31 Publications for this product

Customer reviews and Q&As

1-10 of 17 Abreviews or Q&A

Application
Flow Cytometry
Fixation
Formaldehyde
Permeabilization
Yes - 0.1% PBS-Tween
Sample
Rat Cell (liver)
Specification
liver
Gating Strategy
CD24 CELLS
Preparation
Cell harvesting/tissue preparation method: cell dissociation buffer
Sample buffer: 0.1% PBS BSA

Abcam user community

Verified customer

Submitted Dec 10 2013

Application
Flow Cytometry
Fixation
Formaldehyde
Permeabilization
Yes - BD PERMEABILISATION BUFFER
Sample
Human Cell (HEP G2)
Specification
HEP G2
Gating Strategy
CD24 CELLS
Preparation
Cell harvesting/tissue preparation method: single cell suspension
Sample buffer: 0.1% PBS BSA

Abcam user community

Verified customer

Submitted Dec 06 2013

Application
Western blot
Loading amount
5 µg
Gel Running Conditions
Reduced Denaturing (8)
Sample
Human Cell lysate - whole cell (LIVER)
Specification
LIVER
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C

Abcam user community

Verified customer

Submitted Dec 05 2013

Question
Answer

Gracias por contactarnos.

Espero que no te importe que te conteste en castellano. Recientemente hemos establecido la Oficina Virtual Española y podéis contactarnos en este idioma siempre que lo deseéis. Si preferís que os conteste en inglés, por favor, no dudéis en comunicármelo.

Atendiendo al protocolo, es posible que la membrana se haya bloqueado en exceso al haber diluido ambos anticuerpos en buffer de bloqueo. Además, se ha usado leche para diluir el secundario y BSA para el primario, lo cual añade dos pasos extra de bloqueo al ya realizado antes de incubar con anticuerpo.

Yo os sugeriría diluir los anticuerpos en TBST o PBS en lugar de buffer de bloqueo. Y observar si se obtiene un mejor resultado.

Espero que este consejo permita obtener mejores resultados del anticuerpo. En caso de que no sea así, por favor, no dudéis en contactarnos de nuevo para que podamos ayudaros a resolver el problema lo más rápido y eficazmente posible.

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Answer

Thank you for contacting us.

I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

In order to better understand the problem, I would like to send you a questionnaire with some questions about the protocol used, as well as the order details. All this information is crucial to help us understand the possible causes of the problem. Any images are highly appreciated.

Once we receive the completed questionnaire, we will look at the protocol and see if there are any suggestions we can make that may improve the results. If no suggestions can be made to the protocol, and the antibody was purchased within the last 6 months, it is covered by our Abpromise guarantee, and therefore I can offer you a free of charge replacement or a credit note for it.

I look forward to receiving your reply. Please do not hesitate to contact us if you need any more advice or information.

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Rat Tissue lysate - whole (Rat kidney tissue)
Loading amount
140 µg
Specification
Rat kidney tissue
Gel Running Conditions
Reduced Denaturing (12%)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Aug 30 2012

Answer

Thank you for contacting us. I am sorry that this antibody is detecting multiple bands in WB on rat samples.
Is the secondary antibody you are using pre-adsorbed against rat? It is possible that the anti-mouse secondary may be detecting the heavy and light chains of endogenous IgGs. The heavy and light chains tend to run around 50 and 25 kDa respectively, but variations in buffer system and sample preparation could raise the observed molecular weights and this may account for the bands around 60 and 27 kDa. I would recommend using a preadsorbed secondary antibody, or a secondary antibody that is designed to only detect native IgGs, such as our new VeriBlot range (ab131368 for example).
I would also recommend loading no more than 50 ug of protein to ensure the maximum specificity possible, especially when using a very sensitive ECL reagent.
I hope this helps to improve your results, if not, I will be happy to offer you a free of charge replacement, credit, or refund as long as you have purchased the antibody in the past six months. Please let me know your original order number and how you would like to proceed and I will be happy to help you further.

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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 xxx with ab23331.

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 notcovered by our Abpromise guarantee. Therefore, I am issuing you a testing discount code for testing this replacement antibody in mouse samples. Please see below for details.

I wish you the best of luck with your research.

===== TESTING DISCOUNT CODE =====

DISCOUNT CODE: xxx
Expiration date: xxx

I am very pleased to hear you would like to accept our offer and test ab23331 in mouse. This code will give you 1 free PRIMARY ANTIBODY before the expiration date. To redeem this offer, please submit an Abreview for mouseand 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: https://www.abcam.com/collaborationdiscount.

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Answer

Thank you for calling Abcam. I have continued the search for the guinea pig sequence for stearyl-CoA desaturase, but I have not been able to locate it. I can find, rat, mouse, sheep, goat, bovine, hamster but not guinea pig. Therefore I cannot do an alignment between the human and g.pig sequences to determine likelihood of cross reactivity. If you are still interested in testing this antibody in guinea pig IHC, then I have put the testing discount details below. If there is anything else I can help with, please let me know. Testing Discount Details: As, ab19832 has not been tested in guinea pig I can offer a discount off a future purchase if you buy ab19832 now, test it in (UNTESTED APP/SPECIES) and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of 1 free (PRIMARY ANTIBODY/PROTEIN). If you are interested in this offer, please follow these steps: 1. Reply to this e-mail to let me know that you would like to proceed and test ab19832 in guinea pig. I will then send a discount code. This code must be issued before purchasing ab19832 so please wait for my reply before ordering. 2. Purchase ab19832 either by phone, fax, or online (www.abcam.com). 3. Test it in guinea pig. 4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews. 5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any primary antibody ordered and the discount code is valid for 4 months after issue. We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab19832 turns out to be unsuitable for guinea pig you will still receive the discount on your next purchase after your Abreview has been submitted. Please let me know if you have any questions about this offer and I would be happy to help you further. The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.  

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Question

Dear Sir/Madam, Thanks for your email. I have tried another WB according to your suggestions about the SCD antibody. Unfortunately, the SCD antibody did not work in our lab and I agree that another tube of antibody will not make any difference.  I ran one WB to test the replaced SCD antibody upon I received it, I did use the same lysates I used before and I saw the similar results as before. I ran another WB with your suggestion of using new lysates. However, I had to use the same lysate of NIH 3T3 cells, the positive control.  We don’t have this cell line in our lab and I got one cell pellet from another lab and so now I only have cell lysate. Nevertheless, I got the similar results as all other WB generated from the first SCD antibody or the replaced SCD antibody. Please take a look at the attached file for the results of WB. File SCD-2 is the result from the first SCD antibody with samples loaded onto lane 1-9 as NIH3T3, HepG2, Fibroblast, mouse embryonic fibroblast, mouse liver, rat liver, MCF7, neuroblastma, and Cos7.  File SCD-4 is the result from the replaced antibody with samples loaded onto lane 1-5 as NIH3T3, HepG2, mouse liver, rat liver, and fibroblast.  We normally make fresh lysates, then aliquot, snap-freeze, and kept them at -80oC for future analysis.  For WB analysis, frozen lysates work fine for us and we have been doing this for a long time. Although I can’t think of a reason that a fresh lysate will make the difference, I did re-run anther WB with SCD antibody. Unluckily, the result did not improve. However, for antibody ApoA4 I purchased from Abcam, it is another story. I didn’t saw any specific band for ApoA4 with antibody #ab108200 first. After I stripped the membrane and re-probed it with the replaced antibody of ApoA4 (#ab81616), it gave me the band at the right size and the background was clear.  After so many WB and such a long time trying, I have to say that it clearly does not work in our lab. I was wondering if it is possible that we could get a refund for this antibody. I would greatly appreciate if you could consider the possibility.  Best Regards,  

Read More
Answer

Thank you for your email. We highly appreciate your cooperation in this. I am sorry we are unable to resolve the problems you have been experiencing. It may be possible that these antibodies are not suitable for Typhoon method with Kpi lysis buffer. The ab19862 was cited in 6 publications as far as we know and in 2 publications PubmedID 19787047 and 18697866 in WB so I might suggest checking the protocol they have used. I have asked our account department to refund the money on the original order 912415. The refund ID is 17800. Please contact our customer service team by quoting this order in case of any question. You can also inform your Finance department to contact our Finance department at creditcontrol@abcam.com or by telephone using the information at the “Contact Us” link in the top right corner of our website. 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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1-10 of 17 Abreviews or Q&A

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