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  1. Link

    products/secondary-antibodies/donkey-goat-igg-hl-alexa-fluor-488-ab150129.pdf

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Immunology Immunoglobulins Heavy Chain IgG
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Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) (ab150129)

  • Datasheet
  • SDS
Reviews (6)Q&A (2)References (202)

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Immunocytochemistry/ Immunofluorescence - Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) (ab150129)
  • Flow Cytometry - Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) (ab150129)
  • Alexa Fluor® - Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) (ab150129)

Key features and details

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

Conjugates logo Related conjugates and formulations

Alexa Fluor® 405 Alexa Fluor® 555 Alexa Fluor® 568 Alexa Fluor® 594 Alexa Fluor® 647 Alexa Fluor® 680 Alexa Fluor® 750 Alexa Fluor® 790 Alkaline Phosphatase Biotin Cy2 ® Cy3 ® Cy5 ® Cy5.5 ® DyLight® 488 DyLight® 550 DyLight® 594 DyLight® 650 FITC Gold 10nm Gold 12nm Gold 18nm Gold 25nm HRP HRP polymer PE Texas Red ® TRITC Unconjugated Unconjugated

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Overview

  • Product name

    Donkey Anti-Goat IgG H&L (Alexa Fluor® 488)
    See all IgG secondary antibodies
  • Host species

    Donkey
  • Target species

    Goat
  • Tested applications

    Suitable for: IHC-Fr, ICC/IF, Flow Cyt, IHC-P, ELISAmore details
  • Immunogen

    Full length protein corresponding to Goat IgG.

  • 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 (glycerin, glycerine), 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-Goat
    • IgG
    • Fluorophore
    • Alexa Fluor® 488

Associated products

  • Related Products

    • Normal Donkey Serum (ab7475)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab150129 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 (1)
Use at an assay dependent concentration.
ICC/IF (3)
1/200 - 1/1000.
Flow Cyt
1/2000 - 1/4000.
IHC-P (1)
Use at an assay dependent concentration.
ELISA
Use at an assay dependent concentration.
Notes
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.

Images

  • Immunocytochemistry/ Immunofluorescence - Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) (ab150129)
    Immunocytochemistry/ Immunofluorescence - Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) (ab150129)

    ICC/IF image of ab7291 stained HeLa cells. The cells were 100% methanol fixed (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then incubated in 1%BSA / 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 (ab7291, 1µg/ml) overnight at +4°C. Ab98800, goat anti-mouse IgG, was then added as a secondary bridging antibody, at 1/250 dilution for 1h. Ab150129 Alexa Fluor® 488 donkey anti-goat IgG (H+L) (shown in green) was then used at 1µg/ml for 1h as a tertiary antibody. 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.

  • Flow Cytometry - Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) (ab150129)
    Flow Cytometry - Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) (ab150129)

    Overlay histogram showing Jurkat cells stained Goat polyclonal to Ikaros (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 donkey serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (Goat polyclonal to Ikaros, 0.1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 donkey anti-goat IgG (H&L) (ab150129) at 1/4000 dilution for 30 min at 22°C. Isotype control antibody (black line) was goat IgG (polyclonal) (ab37373, 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.

  • Alexa Fluor® - Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) (ab150129)
    Alexa Fluor® - Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) (ab150129)

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 (202)

Publishing research using ab150129? Please let us know so that we can cite the reference in this datasheet.

ab150129 has been referenced in 202 publications.

  • Stone D  et al. Gene Transfer in Adeno-Associated Virus Seropositive Rhesus Macaques Following Rapamycin Treatment and Subcutaneous Delivery of AAV6, but Not Retargeted AAV6 Vectors. Hum Gene Ther 32:96-112 (2021). PubMed: 32998579
  • Lozic M  et al. Alteration of Cx37, Cx40, Cx43, Cx45, Panx1, and Renin Expression Patterns in Postnatal Kidneys of Dab1-/- (yotari) Mice. Int J Mol Sci 22:N/A (2021). PubMed: 33525532
  • Wang F  et al. aB-Crystallin Alleviates Endotoxin-Induced Retinal Inflammation and Inhibits Microglial Activation and Autophagy. Front Immunol 12:641999 (2021). PubMed: 33777038
  • Li HH  et al. Upregulation of ACE2 and TMPRSS2 by particulate matter and idiopathic pulmonary fibrosis: a potential role in severe COVID-19. Part Fibre Toxicol 18:11 (2021). PubMed: 33706759
  • Huang X  et al. Ckip-1 regulates C3H10T1/2 mesenchymal cell proliferation and osteogenic differentiation via Lrp5. Exp Ther Med 21:342 (2021). PubMed: 33732315
View all Publications for this product

Customer reviews and Q&As

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

1-8 of 8 Abreviews or Q&A

Great secondary antibody anti goat

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
We used low secondary antibody dilution for Green channel. (1:1000)
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 Apr 12 2021

Excellent secondary antibody for FFPE

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
This secondary antibody was used to stain vimentin in human skin sections (nevus).
Concentration: 1:500
Very clear and specific staining
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 Jun 17 2020

Vimentin staining of melanoma cells by indirect IF

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Cell: Sk-Mel28

Fix & permeabilization: Acetone

Blocking: 5% BSA/PBS

1st antibody: anti-Vimentin goat polyclonal antibody (1/100 PBS)

2nd antibody: Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) (ab150129) (1/500 PBS)
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 Jun 01 2020

TIMP1 expression in human skin

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
This antibody was used to study the TIMP1-expression in human skin tissue (melanoma vs nevus) by indirect immunfluorescence.
The anti goat antibody gives very specific and strong signals on frozen-tissue sections with no background. Publishable results.
Concentration 1:200
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 Mar 04 2019

Labeling of granule cells in the Dentate Gyrus of the Mouse Hipocampus

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (PFA fixed)
Very good antibody
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. Yacine Tensaouti

Verified customer

Submitted Aug 01 2017

Brachyury expression in spontaneously differentiated human iPS cells.

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Following fixation in 4% PFA, the cells were assessed for Brachyury (T) expression using 1:100 dilution of the primary antibody in 1% serum, 0.1% triton, 0.1% BSA in PBS, followed by detection using donkey polyclonal rabbit IgG Alexa 488 (ab150129) at 1:500. The results show that nuclear T (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).
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 Sep 02 2013

Question

I want to use your ab150114 and ab150129 to do co-localization test. But I notice that the emission wavelength of these two antibodies overlap quite a lot. Will that affect my result? At the beginning I want to buy Alexa 488 and Alexa 594 to do the experiment, but the problem is our lab don't have the lsm which can detect Alexa 594 (the machine only has laser 405, 488 and 543, and I find the ex of Alexa 594 is 615nm). Could you give me any suggestions? I want to know whether it will work if I am going to buy Alex 488 and 555.

Read More

Abcam community

Verified customer

Asked on Oct 24 2013

Answer

For excitation with the 543nm laser, Alexa Fluor 568 is a better choice than 555 for combination with AF488. There is still some emission spillover but it is much less than for the 488/555 combination, and you should be able to set compensation without as much trouble. If the 488 signal is expected to be much brighter than the other, for instance if that protein is much more abundant than the other, then spillover may still be an issue. For an anti-mouse IgG secondary, I suggest ab175473.

Read More

Tom Ruyle

Abcam Scientific Support

Answered on Oct 24 2013

Question

I am getting significant signal with this antibody diluted 1:1000 in mouse 3T3L1 (preadipocytes) without the addition of any primary antibody. I fix these cells in 4% PFA for 15 minutes and wash 3Xs 10 minutes. Further, I have blocked using 5% donkey serum with 1% BSA, 10% donkey serum, and 10% horse serum for 1 hour at room temperature all with similar results. Any other suggestions?

Read More

Abcam community

Verified customer

Asked on Apr 04 2013

Answer

Having reviewed this case, I would like to offer some suggestions to help optimize the results from ab150129. I would also appreciate if you can confirm some further details: Do you use a detergent such as Tween 20 in your washing buffer? Have you attempted to increase the number and duration of washes after antibody incubation? If so, did this have affect on the background? Have you attempted to further dilute the antibody to 1/1500, 1/2000 or greater? This antibody has not been pre-absorbed against mouse IgG, as such we cannot guarantee that we will be able to eliminate this reactive thought we hope that these suggestions will help reduce this. Abcam does offer a number of AlexaFluor 488 antibodies which have been pre-absorbed to ensure low species cross-reactivity including ab150133 which has been pre-absorbed against chicken, human, mouse, pig, rabbit and rat. More details about this product may be found at: https://www.abcam.com/donkey-polyclonal-secondary-antibody-to-goat-igg-hl-alexa-fluorreg-488-pre-adsorbed-ab150133.html

Read More

Abcam Scientific Support

Answered on Apr 04 2013

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