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Immunology Immunoglobulins Heavy Chain IgG
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Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150073)

  • Datasheet
  • SDS
Reviews (7) Submit a question References (157)

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Immunocytochemistry/ Immunofluorescence - Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150073)
  • Flow Cytometry - Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150073)
  • ELISA - Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150073)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150073)
  • Alexa Fluor® - Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150073)

Key features and details

  • Donkey polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488)
  • Conjugation: Alexa Fluor® 488. Ex: 495nm, Em: 519nm
  • Host species: Donkey
  • Isotype: IgG
  • Suitable for: ICC/IF, Flow Cyt, IHC-P, ELISA, IHC-Fr

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Donkey Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150105)
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Overview

  • Product name

    Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 488)
    See all IgG secondary antibodies
  • Description

    Donkey polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488)
  • Host species

    Donkey
  • Target species

    Rabbit
  • Specificity

    This antibody is specific to Rabbit IgG
  • Tested applications

    Suitable for: ICC/IF, Flow Cyt, IHC-P, ELISA, IHC-Frmore 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. Stable for 12 months at -20°C. Store In the Dark.
  • Storage buffer

    Preservative: 0.02% Sodium azide
    Constituents: 23% Glycerol, PBS, 1% BSA
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Purification notes

    This antibody 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

    • Immunology
    • Immunoglobulins
    • Heavy Chain
    • IgG
    • Secondary antibodies
    • anti-Rabbit
    • IgG
    • Fluorophore
    • Alexa Fluor® 488

Associated products

  • Alternative Versions

    • Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 555) (ab150074)
    • Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 647) (ab150075)
    • Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150076)
    • Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 568) (ab175470)
    • Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 405) (ab175651)
    • Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 750) (ab175731)
    • Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 680) (ab175772)
    • Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 790) (ab175780)
    • Donkey Anti-Rabbit IgG H&L (ab182020)
    • Donkey Anti-Rabbit IgG H&L (HRP) (ab205722)
    • Donkey Anti-Rabbit IgG H&L (Biotin) (ab207999)
  • Related Products

    • Normal Donkey Serum (ab7475)

Applications

Our Abpromise guarantee covers the use of ab150073 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 - 1/4000.
IHC-P Use at an assay dependent concentration.
ELISA Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.

Images

  • Immunocytochemistry/ Immunofluorescence - Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150073)
    Immunocytochemistry/ Immunofluorescence - Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150073)

    ICC/IF image of ab6046 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then incubated in 1% BSA / 10% normal donkey serum / 0.3M glycine in 0.1% PBS-Tween for 1h to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab6046, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab150073 Alexa Fluor® 488 donkey anti-rabbit IgG (H+L) used at 1µg/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.

    This product also gave a positive signal under the same testing conditions in HeLa cells fixed with 4% formaldehyde (10min).

  • Flow Cytometry - Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150073)
    Flow Cytometry - Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150073)

    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 donkey 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 Donkey anti-rabbit IgG H&L (Alexa Fluor® 488) (ab150073) was used at 1/4000 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.

  • ELISA - Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150073)
    ELISA - Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150073)

    Cross-reactivity of the polyclonal secondary antibody ab182020 was tested using a sandwich ELISA approach. The wells were coated with the indicated IgG standards at 1 µg/ml (50 µl/well) and incubated overnight at 4°C, followed by a 5% BSA blocking step for 2h at RT. ab182020 was then added starting at 1 µg/ml and gradually diluted 1/4 (50 µl/well), followed by incubation for 2h. For the detection Goat anti-Donkey IgG H&L (HRP) (ab6988) was used at 1/20,000 dilution (50 µl/well), followed by incubation for 1h at RT.

    For the batch tested, ab182020 showed a cross-reactivity below 2% towards human IgG, mouse IgG, rat IgG, goat IgG and chicken IgY.

    This data was developed using the unconjugated antibody (ab182020).

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150073)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150073)This image is courtesy of an Abreview submitted by Laura Comley.

    ab104603 staining DYNLL1 in mouse cervical spinal cord tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100, and blocked with 10% serum for 1 hour at 21°C. Samples were incubated with primary antibody (2 µg/ml in PBS/10% serum/0.1% Triton X-100) for 16 hours at 4°C. An Alexa Fluor® 488-conjugated Donkey anti-rabbit IgG H&L (ab150073) (1/500) was used as the secondary antibody.

  • Alexa Fluor® - Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150073)
    Alexa Fluor® - Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150073)

Protocols

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

Datasheets and documents

    • Datasheet
    • SDS
  • References (157)

    Publishing research using ab150073? Please let us know so that we can cite the reference in this datasheet.

    ab150073 has been referenced in 157 publications.

    • Chen M  et al. The plant natural product 2-methoxy-1,4-naphthoquinone stimulates therapeutic neural repair properties of olfactory ensheathing cells. Sci Rep 10:951 (2020). PubMed: 31969642
    • Walkden H  et al. Burkholderia pseudomallei invades the olfactory nerve and bulb after epithelial injury in mice and causes the formation of multinucleated giant glial cells in vitro. PLoS Negl Trop Dis 14:e0008017 (2020). PubMed: 31978058
    • Craver BM  et al. N-acetylcysteine inhibits thrombosis in a murine model of myeloproliferative neoplasm. Blood Adv 4:312-321 (2020). PubMed: 31978215
    • Li J  et al. Long non-coding RNA X-inactive specific transcript silencing ameliorates primary graft dysfunction following lung transplantation through microRNA-21-dependent mechanism. EBioMedicine 52:102600 (2020). PubMed: 31981974
    • Hong YP  et al. High-Fat Diet Aggravates Acute Pancreatitis via TLR4-Mediated Necroptosis and Inflammation in Rats. Oxid Med Cell Longev 2020:8172714 (2020). PubMed: 31998444
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-7 of 7 Abreviews or Q&A

    FACS detection of CAT1 on HCT116 cells

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Flow Cytometry
    Sample: CAT1-overexpressed HCT116 cells
    Condition: No fixation, No permeabilization
    1st antibody: Rabbit anti-CAT1 antibody (ab37588) or Rabbit control IgG
    2nd antibody: anti-Rabbit IgG Alexa-488 (ab150073)

    Result: CAT1 was specifically detected by FACS.

    Abcam user community

    Verified customer

    Submitted Aug 22 2017

    Donkey A488 anti-Rabbit

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (PFA fixed)
    Works very well, I recommend

    Yacine Tensaouti

    Verified customer

    Submitted Jul 26 2017

    immunofluorescent staining of iNOS in mouse hepatocytes.

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunomicroscopy
    Donkey anti-Rabbit IgG H&L (Alexa Fluor® 488) is nice to label the iNOS in mouse hepatocytes.

    Abcam user community

    Verified customer

    Submitted Aug 07 2015

    ICC of Mouse brain organotypic slices

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry
    This secondary marker was used at a dilution of 1/500 with PBS for 2 hours in combination with a primary H2Ax nuclear anti-body and the nuclear stainTOPRO3 in brain tissue from mice

    Ms. Kirsty Ireland

    Verified customer

    Submitted Mar 18 2014

    Human Nanog expression vector in WT mouse embryonic stem cells.

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Nice, bright clear staining seen using secondary antibody at a 1:200 dilution. Primary antibody used here was anti-Nanog (ab21624). Small amounts of speckled areas noted.

    Dr. Sarah Ritson

    Verified customer

    Submitted Oct 07 2013

    Western blot abreview for Anti-GAPDH antibody

    Inconclusive
    Abreviews
    Abreviews
    Application
    Western blot
    Application Western blot
    Sample Mouse Rat lysate - Cytoplasm
    Loading amount 50 µg
    Gel Running Conditions Reduced Denaturing (10)
    Blocking step Milk as blocking agent for 1 hour(s) and 30 minute(s)· Concentration: 5% · Temperature: 25°C
    Other product details
    Primary antibody- ab17820 Rabbit Polyclonal to Caspase 1
    Dilution 1/1000, 1/2000. 1/5000
    Incubation time 24 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: TBST
    Secondary antibody
    Application Western blot
    Dilution 1/1000, 1/2000, 1/5000
    Detection
    Biorad Gel Doc XR+
    Bands Specific: 45, 20 kDa
    Stained membrane to check for protein transfer, all looked good. Incubated in both primary and secondary antibody and imaged, no bands were present at any of the dilutions.

    Ms. Tammie Quinn

    Verified customer

    Submitted Sep 26 2013

    Expression of pluripotency marker Sox-2 in human ES cells.

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Following fixation in 4% PFA, the cells were assessed for Sox2 expression using 1:100 dilution of the primary antibody (ab97959) in 1% serum, 0.1% triton, 0.1% BSA in PBS, followed by detection using donkey polyclonal rabbit IgG Alexa 488 (ab150073) at 1:500. The results show that nuclear Sox-2 (green) was clearly observed. An isotype control IgG was run in parallel and showed no positive staining (not shown here). All nuclei were counterstained using DAPI (blue).

    Abcam user community

    Verified customer

    Submitted Sep 02 2013

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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