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    donkey-rabbit-igg-hl-alexa-fluor-405-ab175651.pdf

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Immunology Immunoglobulins Heavy Chain IgG
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Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 405) (ab175651)

  • Datasheet
  • SDS
Reviews (3) Submit a question References (35)

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Immunohistochemistry (Frozen sections) - Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 405) (ab175651)
  • Immunocytochemistry/ Immunofluorescence - Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 405) (ab175651)
  • ELISA - Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 405) (ab175651)
  • Alexa Fluor® - Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 405) (ab175651)

Key features and details

  • Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 405)
  • Conjugation: Alexa Fluor® 405. Ex: 402nm, Em: 421nm
  • Host species: Donkey
  • Isotype: IgG
  • Suitable for: IHC-Fr, Flow Cyt, IHC-P, ELISA, ICC/IF

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 568 Alexa Fluor® 594 Alexa Fluor® 647 Alexa Fluor® 680 Alexa Fluor® 750 Alexa Fluor® 790 Alkaline Phosphatase Biotin DyLight® 488 DyLight® 550 DyLight® 594 DyLight® 650 FITC Gold 10nm Gold 12nm Gold 18nm Gold 25nm Gold 6nm HRP IRDye® 680RD Texas Red ® TRITC Unconjugated Unconjugated

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Overview

  • Product name

    Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 405)
    See all IgG secondary antibodies
  • Host species

    Donkey
  • Target species

    Rabbit
  • Tested applications

    Suitable for: IHC-Fr, Flow Cyt, IHC-P, ELISA, ICC/IFmore details
  • Immunogen

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

  • Conjugation

    Alexa Fluor® 405. Ex: 402nm, Em: 421nm

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 (glycerin, glycerine), PBS, 1% BSA
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • General notes

    We recommend storage time at 4°C should be minimal, since this may affect the signal strength.

    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® 405

Associated products

  • Affibody® Molecule

    • Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150063)
  • Alternative Versions

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

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab175651 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.
Flow Cyt
1/2000.
IHC-P
Use at an assay dependent concentration.
ELISA
Use at an assay dependent concentration.
ICC/IF
1/200 - 1/1000.

We recommend the use of a dedicated 405 filter for optimal results not the DAPI filter. The DAPI filter may not excite until the maximum emission peaks of Alexa Fluor® 405 dye (see difference below) 
Ex max: Alexa Fluor® 405 = 402nm / DAPI = 359nm 
Em max: Alexa Fluor® 405 = 421nm / DAPI = 461nm

Notes
IHC-Fr
Use at an assay dependent concentration.
Flow Cyt
1/2000.
IHC-P
Use at an assay dependent concentration.
ELISA
Use at an assay dependent concentration.
ICC/IF
1/200 - 1/1000.

We recommend the use of a dedicated 405 filter for optimal results not the DAPI filter. The DAPI filter may not excite until the maximum emission peaks of Alexa Fluor® 405 dye (see difference below) 
Ex max: Alexa Fluor® 405 = 402nm / DAPI = 359nm 
Em max: Alexa Fluor® 405 = 421nm / DAPI = 461nm

Images

  • Immunohistochemistry (Frozen sections) - Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 405) (ab175651)
    Immunohistochemistry (Frozen sections) - Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 405) (ab175651)Image courtesy of an Abreview submitted by Daniel Coutu.

    Section of adult murine femur was stained with rabbit anti-fibronectin (ab23750) and detected with donkey anti-rabbit AlexaFluor405 (ab175651) - shown in green. Counterstain shows CD31+ blood vessels and cells (red) and Sca1+ arterioles and cells (blue).

  • Immunocytochemistry/ Immunofluorescence - Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 405) (ab175651)
    Immunocytochemistry/ Immunofluorescence - Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 405) (ab175651)

    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, blocked with Image-iT® FX Signal Enhancer at room temperature for 30 min 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 (blue) was ab175651 Alexa Fluor® 405 donkey anti-rabbit IgG (H+L) used at 2µg/ml for 1h. DRAQ5 (ab108410) was used to stain the cell nuclei (red) at a concentration of 1.25µM.

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

  • ELISA - Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 405) (ab175651)
    ELISA - Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 405) (ab175651)

    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).

     

  • Alexa Fluor® - Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 405) (ab175651)
    Alexa Fluor® - Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 405) (ab175651)

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

  • SDS download

  • Datasheet download

    Download

References (35)

Publishing research using ab175651? Please let us know so that we can cite the reference in this datasheet.

ab175651 has been referenced in 35 publications.

  • Clawson BC  et al. Causal role for sleep-dependent reactivation of learning-activated sensory ensembles for fear memory consolidation. Nat Commun 12:1200 (2021). PubMed: 33619256
  • Alday-Parejo B  et al. MAGI1 localizes to mature focal adhesion and modulates endothelial cell adhesion, migration and angiogenesis. Cell Adh Migr 15:126-139 (2021). PubMed: 33823745
  • Sun B  et al. Regulation of host and virus genes by neuronal miR-138 favours herpes simplex virus 1 latency. Nat Microbiol 6:682-696 (2021). PubMed: 33558653
  • Sato H  et al. Metabolome Shift in Both Metastatic Breast Cancer Cells and Astrocytes Which May Contribute to the Tumor Microenvironment. Int J Mol Sci 22:N/A (2021). PubMed: 34299050
  • Tona Y & Wu DK Live imaging of hair bundle polarity acquisition demonstrates a critical timeline for transcription factor Emx2. Elife 9:N/A (2020). PubMed: 32965215
View all Publications for this product

Customer reviews and Q&As

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

1-3 of 3 Abreviews or Q&A

Wholemount IF (PFA)

Excellent
Abreviews
Abreviews
abreview image
Application
Other - Do not use
Reliably produces clean signal without aggregates or precipitates even at 1:100 dilution.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jan 27 2020

Microglia in brain slice cultures

Good
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry
Stained with anti-IBA1 antibody and Alexa 405 - counter stained with TOPRO3
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Feb 01 2016

Donkey anti-rabbit AlexaFluor405 for detection of fibronectin in murine bone marrow

Good
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Section of adult murine femur was stained with rabbit anti-fibronectin (ab23750) and detected with donkey anti-rabbit AlexaFluor405 (ab175651) - shown in green. Counterstain shows CD31+ blood vessels and cells (red) and Sca1+ arterioles and cells (blue).
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Daniel Coutu

Verified customer

Submitted Mar 28 2014

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