Rabbit Anti-Sheep IgG H&L (FITC) (ab6743)

Reviews (1)Q&A (5)References (10)

Key features and details

  • Rabbit Anti-Sheep IgG H&L (FITC)
  • Conjugation: FITC. Ex: 493nm, Em: 528nm
  • Host species: Rabbit
  • Suitable for: Immunomicroscopy, Flow Cyt, ICC/IF, IHC-P, IHC-Fr, ELISA

製品の概要

  • 製品名

    Rabbit Anti-Sheep IgG H&L (FITC)
    IgG 二次抗体 製品一覧

  • 由来種

    Rabbit

  • ターゲット生物種

    Sheep

  • アプリケーション

    適用あり: Immunomicroscopy, Flow Cyt, ICC/IF, IHC-P, IHC-Fr, ELISAmore details

  • 免疫原

    Sheep whole molecule

  • 標識

    FITC. Ex: 493nm, Em: 528nm

製品の特性

  • 製品の状態

    Liquid

  • 保存方法

    Shipped at 4°C. Store at +4°C.

  • バッファー

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 0.878% Sodium chloride, 1% BSA, 0.424% Potassium phosphate
  • Concentration information loading...

  • 精製度

    Affinity purified

  • 特記事項（精製）

    This product was prepared from monospecific antiserum by immunoaffinity chromatography using Sheep IgG coupled to agarose beads followed by solid phase adsorption(s) to remove any unwanted reactivities.

  • 特記事項（標識）

    This product was prepared from monospecific antiserum by immunoaffinity chromatography using Sheep IgG coupled to agarose beads followed by solid phase adsorption(s) to remove any unwanted reactivities.

  • ポリ/モノ

    ポリクローナル

  • 特記事項

    Diaminotriazinylaminofluorescein (DTAF) (Molecular Weight 530 daltons) Absorption Wavelength: 495 nm Emission Wavelength: 528 nm Fluorochrome/Protein Ratio: 2.3 moles DTAF per mole of Rabbit IgG.

  • 研究分野

アプリケーション

Our Abpromise guarantee covers the use of ab6743 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
Immunomicroscopy Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent dilution.
ICC/IF 1/1000 - 1/5000.
IHC-P Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
ELISA 1/10000 - 1/50000.

プロトコール

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

参考文献 (10)

ab6743 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab6743 は 10 報の論文で使用されています。

  • Zhou QL  et al. FPR1 gene silencing suppresses cardiomyocyte apoptosis and ventricular remodeling in rats with ischemia/reperfusion injury through the inhibition of MAPK signaling pathway. Exp Cell Res 370:506-518 (2018). PubMed: 30031130
  • Mendoza JL  et al. The IFN-?-IFN-?R1-IL-10Rß Complex Reveals Structural Features Underlying Type III IFN Functional Plasticity. Immunity 46:379-392 (2017). PubMed: 28329704
  • Tang J  et al. Construction of a general albumin promoter reporter system for real-time monitoring of the differentiation status of functional hepatocytes from stem cells in mouse, rat and human. Biomed Rep 6:627-632 (2017). PubMed: 28584633
  • Wilding C  et al. ?-Synuclein antibodies have neuroprotective potential on neuroretinal cells via proteins of the mitochondrial apoptosis pathway. PLoS One 9:e90737 (2014). PubMed: 24595072
  • Nguyen MD  et al. Cardiac cell culture model as a left ventricle mimic for cardiac tissue generation. Anal Chem 85:8773-9 (2013). PubMed: 23952579
  • Pippin JW  et al. Cells of renin lineage are progenitors of podocytes and parietal epithelial cells in experimental glomerular disease. Am J Pathol 183:542-57 (2013). PubMed: 23769837
  • Li G  et al. Beneficial effects of a novel RAGE inhibitor on early diabetic retinopathy and tactile allodynia. Mol Vis 17:3156-65 (2011). PubMed: 22171162
  • Hauser PV  et al. Novel siRNA delivery system to target podocytes in vivo. PLoS One 5:e9463 (2010). PubMed: 20209128
  • Chiriva-Internati M  et al. AKAP-4: a novel cancer testis antigen for multiple myeloma. Br J Haematol 140:465-8 (2008). PubMed: 18217892
  • Shah MD  et al. Flow cytometric measurement of microparticles: pitfalls and protocol modifications. Platelets 19:365-72 (2008). PubMed: 18791943

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1-6 of 6 Abreviews or Q&A

For IHC-P

 Excellent
Abreviews
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
I tried 1:100 and 1:500 dilutions for this antibody. 1:500 works well with an hour incubation at room temperature.

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Verified customer

投稿 Aug 20 2014

Question

Please let me know the molecular weight of those antibodies.

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Answer

Thank you for your inquiry.

Unfortunately, we do not determine the exact molecular weight for our antibodies.

https://www.abcam.com/ab6743: asshown on the datasheet,the fluorochrome/Protein Ratio is2.3 moles DTAF per mole of Rabbit IgG.

The molecular weight of DTAF is 495 g/mol and the molecular weight of an IgG moleculeis about 150 kDa. The molecular weight of one molecule of ab6743 is therefore about 151kDa.(=2.3x495+150000).

I hope this is helpful.

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Question

Finalmente como abes he comprado ab8940 anti-human serum albumin

y ab6743 anti sheep, la primera prueba no ha sido demasiado exitosa,

por lo que quería preguntar, si teneis algun protocolo de refencia.

Muchas gracias por vuestra atención

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Answer

Gracias por tu respuesta.

No tenemos un protocolo concreto para el uso de este anticuerpo mediante immunohistoquímica, sino protocolos generales que pueden servir de guía para los distintos ensayos.

Te adjunto el link del protocolo de immunohistoquímica para secciones incluidas en parafina:

https://www.abcam.com/index.html?pageconfig=resource&rid=11384

Así como el “recetario” de los distintos pasos a seguir:

https://www.abcam.com/index.html?pageconfig=resource&rid=13046

También te pueden resultar útiles los consejos para tratar los posibles problemas que pueden darse a lo largo del ensayo:

https://www.abcam.com/index.html?pageconfig=resource&rid=11467

Las únicas recomendaciones concretas para el uso de estos anticuerpos es utilizar como agente de bloqueo suero de conejo (especie de procedencia del secundario) y usar el primario a una dilución de 1/1000. Quizás te ayude echar un vistazo al Abreview publicado por otro usuario del anticuerpo ab8940 para comprobar en qué condiciones usó el anticuerpo.

Es importante también usar siempre controles positivos para ayudar a determinar el origen de los posibles problemas, así como llevar a cabo controles no primarios (incubando únicamente el secundario).

En cualquier caso, ambos anticuerpos están cubiertos por nuestra garantía Abpromise, y si siguieras teniendo problemas con ellos te ruego que nos contactes para ayudarte a resolverlos.

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Question

Muchas gracias por tus comentarios.
Como sistema de recuperación antigénica utilizo normalmente citrato sódico y microondas.
Me comentas lo de usar suero de mismo secundario, pero en este caso no seri sheep?

Muchas gracias

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Answer

Gracias por tu respuesta.
Te confirmo que ab53435 solo se ha testado en secciones congeladas. No tenemos, pues, ninguna evidencia de que vaya a detectar el epítopo en secciones incluidas en parafina.
Así pues, puedes optar por testarlo en esta aplicación, indicándomelo previamente para que yo te envíe el código canjeable por un anticuerpo gratuito, u optar por usar ab8940.
Si eliges la última opción te aconsejo que uses el método de recuperación antigénica que te suele funcionar. Yo particularmente aconsejo Tris EDTA en microondas, optimizando el tiempo de recuperación, es decir, probando a varios tiempos, a partir de un minuto, y no más de 20, para evitar dañar la morfología del tejido.
Como agente de bloqueo, la recomendación es usar suero de la misma especie en la que se generó el secundario, para evitar uniones inespecíficas de éste. Si utilizas el Rabbit anti-Sheep IgG H&L (FITC) secondary antibody (ab6743), lo ideal sería usar suero de conejo.
Espero haberte ayudado. Recuerda que si optas por probar ab53435 en IHC-P, el código debe emitirse en primer lugar, por lo que antes de hacer el pedido te pido que me lo indiques para enviarte dicho código.
Si tienes alguna otra consulta, no dudes en volver a contactarnos.

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Question

I have received ab6743 from you and have been using it as the secondary ab against a sheep anti-histone ab (Biogenesis catalog #4974-8060). I am testing N18 cells (an adherent mouse neuroblastoma cell line) to determine if histones are ever expressed on the cell surface. (Likely an artifact but the point of doing this is to confirm that it's not true). For my first attempt I used the Cytofix/Cytoperm kit by Pharmingen to permeabalize the cells. A fraction of a percent of the cells lit up. I have done this experiment2 more times using 0.4% Triton x-100 (after paraformaldehdye fixation) to permeabalize cells. None of the cells have lit up. What do you recommend for permeabilization of adherrent cells? Also, have you confirmed your ab works in this type of experiment? or with the Biogenesis ab?

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Answer

We have no experience using this secondary antibody in this technique, so I am afraid that I cannot offer any specific advice. However, this Rabbit polyclonal to sheep IgG will certainly work in conjunction with a sheep primary.

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Question

I have received your antibodies, ab6743 and ab7834, and would like to know if they are stable in buffers containing 0.4% Triton X-100. I need to permeabilize cells and the protocol I intend to use calls for fixation with 4% paraformaldehyde in PBS followed by permeabilization with PBS containing 0.4% Triton X-100, 1% BSA and 4% normal serum. Will your antibodies be ok in this buffer? Thanks, Ron Knight

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Answer

Antibodies are usually stable in buffers containing triton-X. I've personally used antibodies in such buffers with no problems. Also, several automated IHC staining systems routinely use such surfactants in their buffer solutions. We have never actually tested ab6743 and 7834 in Triton-X containing buffers, but the odds are that they will be OK. Try a small amount for starters to make sure.

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