Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)

製品の概要

  • 製品名
    Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed
    IgG 二次抗体 製品一覧
  • 製品の詳細
    Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed
  • 由来種
    Goat
  • ターゲット生物種
    Rabbit
  • 特異性

    By immunoelectrophoresis and ELISA this antibody reacts specifically with rabbit IgG and with light chains common to other rabbit immunoglobulins. No antibody was detected against non-immunoglobulin serum proteins. Reduced cross-reactivity to bovine, chicken, horse, human, mouse, pig, and rat IgG was detected. This antibody may cross react with IgG from other species.

     

  • アプリケーション
    適用あり: IHC-Fr, ICC/IF, Flow Cyt, IHC-P, ELISAmore details
  • 吸着処理血清

    Human, Mouse, Rat more details
  • 免疫原

    The details of the immunogen for this antibody are not available.

  • 標識
    Alexa Fluor® 488. Ex: 495nm, Em: 519nm

製品の特性

  • 製品の状態
    Liquid
  • 保存方法
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • バッファー
    Preservative: 0.02% Sodium azide
    Constituents: 1% BSA, 30% Glycerol, PBS
  • Concentration information loading...
  • 精製度
    Immunogen affinity purified
  • 特記事項(精製)
    Antiserum was cross adsorbed using a human, mouse and rat immunosorbents to remove cross reactive antibodies. This antibody was isolated by affinity chromatography using antigen coupled to agarose beads.
  • ポリ/モノ
    ポリクローナル
  • アイソタイプ
    IgG
  • 特記事項

    Alexa Fluor® is a registered trademark of Molecular Probes, Inc, a Thermo Fisher Scientific Company. The Alexa Fluor® dye included in this product is provided under an intellectual property license from Life Technologies Corporation. As this product contains the Alexa Fluor® dye, the purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). As this product contains the Alexa Fluor® dye the sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: in manufacturing; (ii) to provide a service, information, or data in return for payment (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are sold for use in research. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, 5781 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@thermofisher.com.

  • 研究分野

アプリケーション

Our Abpromise guarantee covers the use of ab150081 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
IHC-Fr Use at an assay dependent concentration.
ICC/IF 1/200 - 1/1000.
Flow Cyt 1/2000 - 1/4000.
IHC-P Use at an assay dependent concentration.
ELISA Use at an assay dependent concentration.

画像

  • ICC/IF image of ab8227 stained HeLa cells. The cells were 4% formaldehyde 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 (ab8227, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab150081 Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at 2µg/ml for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    The negative control (inset) is a secondary-only assay to demonstrate low non-specific binding of the secondary antibody.

  • Overlay histogram showing Jurkat cells stained with ab16669 (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 (ab16669, 1/1000 dilution) for 30 min at 22°C. The secondary antibody Goat anti-rabbit IgG H&L (Alexa Fluor® 488, pre-adsorbed) (ab150081) was used at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
  • ab92742 staining Ki67 in wild-type HAP1 cells (top panel) and Ki67 knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab92742 at 1µg/ml and ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 µg/ml (shown in green). Nuclear DNA was labeled in blue with DAPI.

  • Unpurified ab134175 staining Cyclin D1 in MCF7 (Human breast adenocarcinoma cell line) cells treated with KN-93 (ab120980).
    The cells were fixed with 100% methanol (5min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab134175 at 10µg/ml and ab7291 at 1µg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with an Goat anti-Rabbit Alexa 488 secondary (ab150081) at 2 µg/ml (shown in green) and Goat anti-Mouse Alexa 594 secondary (ab150120) at 2 µg/ml (shown in pseudo color red). Nuclear DNA was labeled in blue with DAPI.
    Negative controls: 1- Rabbit primary and anti-mouse secondary antibody; 2 - Mouse primary antibody and anti-rabbit secondary antibody. Controls 1 and 2 indicate that there is no unspecific reaction between primary and secondary antibodies used.

  • Postnatal day 6 mouse testes were fixed with 4% paraformaldehyde. Tissue was embedded in O.C.T. and frozen. 5 micron sections were cut and transferred to slides. Sections were permeabilized with 0.1% Triton X-100 in PBS, and blocked with 3% BSA in 0.1% Triton X-100 + PBS. Sections were incubated with either (A) no primary antibody or (B ) anti-DDX4 (ab13840) for 1 h at RT. Sections were then washed 3X with 0.1% Triton X-100 in PBS and Goat-Anti Rabbit 488 (ab150081) applied at a 1/500 dilution. Sections were then mounted after washing 3X with 0.1% Triton X-100 in PBS.

  • Image: Courtesy of Dr. Shaohua Li, UMDNJ-Robert Wood Johnson Medical School

    Sample: mouse embryonic stem cell-differentiated embryoid bodies (EBs)

    Preparation:

    Fix in 3%PFA in PBS for 30 min at RTIncubate in 7.5% sucrose-PBS for 3h at RTIncubate in 15% sucrose-PBS at 4 degree Celsius overnightEmbed the EBs in tissue-Tek OCT compoundCut frozen sections to 4-20 µm thickness

    Primary antibody 1: Rabbit anti cytokeratin 8 (ab53280), 1:100

    Primary antibody 2: Rat anti-perlecan, 1:100 
    Secondary antibody 1: Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) pre-adsorbed (ab150081), 1:200

    Secondary antibody 2: Goat polyclonal Secondary Antibody to Rat IgG - H&L (Cy5®) pre-adsorbed, 1:200
    Nuclei were counterstained with DAPI 

参考文献

This product has been referenced in:
See all 18 Publications for this product

レビューと Q&A

1-4 of 4 Abreviews or Q&A

Abreviews
Application
Immunocytochemistry/ Immunofluorescence
We used the antibody together with the ab38578 antibody (Siglec8) on cytospins with eosinophilic granulocytes.
The working concentration was 1/100 for 30 minutes at room temperature.

Abcam user community

Verified customer

投稿 Feb 14 2018

Abreviews
Application
Immunohistochemistry (Frozen sections)
I have used this antibody at a 1:500 dilution with many different primary antibodies. It works well and and gives a bright signal with no discernible nonspecific staining.
Postnatal day 6 mouse testes were fixed with 4% paraformaldehyde. Tissue was embedded in O.C.T. and frozen. 5 micron sections were cut and transferred to slides. Sections were permeabilized with 0.1% Triton X-100 in PBS, and blocked with 3% BSA in 0.1% Triton X-100 + PBS. Sections were incubated with either (A) no primary antibody or (B ) anti-DDX4 (Abcam ab13840) for 1 h at RT. Sections were then washed 3X with 0.1% Triton X-100 in PBS and Goat-Anti Rabbit 488 (Abcam ab150081) applied at a 1:500 dilution. Sections were then mounted after washing 3X with 0.1% Triton X-100 in PBS.

Mr. Bryan Niedenberger

Verified customer

投稿 Mar 09 2016

Application
Immunocytochemistry/ Immunofluorescence
This antibody has been used extensively with a number of different primary antibodies generated in rabbits. It performs extremely well at a 1:200 dilution.
Attached is an immunofluorescent image of MeOH-fixed HeLa cells that were immunofluorescently labeled with a Lamin A/C (nuclear membrane) antibody (ab133256). The secondary antibody has been pseudo-colored green in the merged image. DNA was counterstained with DAPI.

Dr. Kirk Mcmanus

Verified customer

投稿 Mar 28 2014

Application
Immunohistochemistry
Application: Immunohistochemistry (PFA perfusion fixed frozen sections) or IHC(FoFr)

I used this antibody on PFA fixed, sucrose cryoprotected frozen sections from the mouse brain (below is a brief description). The primary a.b. I used was anti-mGluR5 (ab53090), which was somewhat difficult to get working.

Full Description:
Transcardially perfused the mouse with 4% PFA. Dissected the brain and placed in PFA for a few hours later, placed in 30% sucrose overnight for cryoprotection. Then, snap/flash froze the brain and used a cryostat for sectioning into 20 micron coronal slices. Applied primary anti-mGluR5 (ab53090) antibody, followed by the secondary antibody (dilution: 1/200), according to standard protocols. No blocking, no antigen-retrieval, and no permeabilization was done.

Abcam user community

Verified customer

投稿 Jan 29 2014

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