Overview

  • Product name
    Goat Anti-Rat IgG H&L (Alexa Fluor® 488) preadsorbed
    See all IgG secondary antibodies
  • Description
    Goat polyclonal Secondary Antibody to Rat IgG - H&L (Alexa Fluor® 488), pre-adsorbed
  • Host species
    Goat
  • Target species
    Rat
  • Specificity
    By immunoelectrophoresis and ELISA this antibody reacts specifically with rat IgG and with light chain common to other rat immunoglobulins. No antibody was detected against non-immunoglobulin serum proteins. Less than 1% cross reactivity to bovine, chicken, human, mouse, rabbit and sheep IgG was detected. This antibody may cross react with IgG from other species.
  • Tested applications
    Suitable for: ICC/IF, Flow Cyt, ELISA, IHC-P, IHC-Frmore details
  • Minimal
    cross-reactivity

    Chicken, Cow, Human, Mouse, Rabbit, Sheep more details
  • Immunogen

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

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

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • Storage buffer
    Preservative: 0.02% Sodium azide
    Constituents: 1% BSA, 30% Glycerol, PBS
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Purification notes
    Antiserum was cross adsorbed using bovine, chicken, human, mouse, rabbit and sheep immunosorbents to remove cross reactive antibodies. The antibody to rat IgG was isolated by affinity chromatography using antigen coupled to agarose beads.
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • General notes

    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.

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab150165 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 1/200 - 1/1000.
Flow Cyt 1/2000.
ELISA Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.

Images

  • ab6326 stained in Hela cells. Untreated and BrdU treated (10uM for 24 hours) cells were fixed with 100% methanol (5 min) and then subjected to acid hydrolysis using 2M HCL in 0.1% PBS-Tween for 30 minutes at room temperature to denature the DNA. They were then incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% triton for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab6326 at 5µg/ml and ab7291 (Mouse monoclonal to alpha tubulin) at 1ug/ml overnight at +4°C. The secondary antibodies were Goat Anti-Rat IgG H&L (Alexa Fluor® 488) preadsorbed (ab150165) (colored green) used at 2 ug/ml and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) (pseudo-colored red) used at 1/1000 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43µM for 1hour at room temperature.

  • ICC/IF image of ab6160 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 (ab6160, 2µg/ml) overnight at +4°C. The secondary antibody (green) was ab150165 Alexa Fluor® 488 goat anti-rat IgG (H&L) used at 2µg/ml for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
  • Overlay histogram showing HeLa cells stained with ab19136 (red line). The cells were fixed with 80% methanol (5 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 (ab19136, 1μg/1x10^6 cells) for 30 min at 22ºC. The secondary antibody used was Alexa Fluor® 488 goat anti-rat IgG (H&L) at 1/2000 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rat IgG2a [aRTK2758] (ab18450, 2µg/1x10^6 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.

  • Dot plot showing BrdU-treated HeLa cells stained with ab6326. Cells were incubated with 10 µM BrdU for 30 minutes prior to being harvested, washed twice in 1x PBS and fixed in 70% ethanol (4°C, added drop-wise) for at least 30 minutes on ice. Once fixed, pellets were acid denatured with 2M HCl for 30 minutes at 22°C and then neutralised with borate buffer (0.1M, pH8.5).
    Samples were washed and incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab6326, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was Goat Anti-Rat IgG H&L (Alexa Fluor® 488) preadsorbed (ab150165) at 1/2000 dilution for 30 min at 22°C.
    7-AAD was added to cells 20 min prior to data acquisition.
    Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) with 530/30 and 685/35 bandpass filters.

  • 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-KIT (ab65525) for 1 h at RT. Sections were then washed 3X with 0.1% Triton X-100 in PBS and Goat-Anti Rat 488 (ab150165) applied at a 1/500 dilution. Sections were then mounted after washing 3X with 0.1% Triton X-100 in PBS.

  • HeLa cells showing negative staining by ICC/IF using only secondary antibody. 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 secondary antibody (green) was ab150165 Alexa Fluor® 488 goat anti-rat IgG (H&L) used at 2µg/ml for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

References

This product has been referenced in:
  • McMahon D  et al. Angiogenic response of rat hippocampal vasculature to focused ultrasound-mediated increases in blood-brain barrier permeability. Sci Rep 8:12178 (2018). Read more (PubMed: 30111814) »
  • Nickel S  et al. Expression and Activity of Breast Cancer Resistance Protein (BCRP/ABCG2) in Human Distal Lung Epithelial Cells In Vitro. Pharm Res N/A:N/A (2017). Read more (PubMed: 28470471) »
See all 2 Publications for this product

Customer reviews and Q&As

Abreviews
Application
Immunohistochemistry (Frozen sections)
I have used this antibody at a 1:500 dilution with rat anti-KIT as a primary antibody. It works well and and gives a produces a bright signal with low backround.
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-KIT (Abcam ab65525) for 1 h at RT. Sections were then washed 3X with 0.1% Triton X-100 in PBS and Goat-Anti Rat 488 (Abcam ab150165) 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

Submitted Mar 29 2016

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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