Goat Anti-Mouse IgG H&L (DyLight® 488) (ab96871)

Reviews (3)Q&A (9)References (9)

Overview

  • Product name
    Goat Anti-Mouse IgG H&L (DyLight® 488)
    See all IgG secondary antibodies
  • Host species
    Goat
  • Target species
    Mouse
  • Specificity
    By immunoelectrophoresis and ELISA this antibody reacts specifically with Mouse IgG and with light chains common to other Mouse immunoglobulins. No antibody was detected against non immunoglobulin serum proteins.
  • Tested applications
    Suitable for: IHC-P, ICC/IF, Flow Cytmore details
  • Conjugation
    DyLight® 488. Ex: 493nm, Em: 518nm

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C.
  • Storage buffer
    Preservative: 0.09% Sodium azide
    Constituents: 0.2% BSA, PBS
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Purification notes
    This antibody was isolated by affinity chromatography using antigen coupled to agarose beads and conjugated to DyLight® 488.
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab96871 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-P 1/50 - 1/500.
ICC/IF 1/50 - 1/500.
Flow Cyt 1/1000 - 1/2000.

Images

  • Flow Cytometry - Goat Anti-Mouse IgG H&L (DyLight® 488) (ab96871)
    Flow Cytometry - Goat Anti-Mouse IgG H&L (DyLight® 488) (ab96871)
    Overlay histogram showing Jurkat cells stained with ab8090 (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 (ab8090, 0.01μg/1x106 cells) for 30 min at 22°C. The secondary antibody Goat anti-mouse IgG H&L (DyLight® 488) (ab96871) was used at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 0.01μg/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.

References

This product has been referenced in:
  • Langhauser F  et al. A diseasome cluster-based drug repurposing of soluble guanylate cyclase activators from smooth muscle relaxation to direct neuroprotection. NPJ Syst Biol Appl 4:8 (2018). Read more (PubMed: 29423274) »
  • Gui Z  et al. Oridonin inhibition and miR-200b-3p/ZEB1 axis in human pancreatic cancer. Int J Oncol 50:111-120 (2017). Read more (PubMed: 27878247) »
See all 9 Publications for this product

Publishing research using ab96871? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-10 of 12 Abreviews or Q&A

Immunocytochemistry (primary skin fibroblast cells)

 Excellent
Abreviews
abreview image
Application
Immunocytochemistry
1. Fixation: 95% ethanol for 10 min
2. Washing: 1x PBS 3 min
3. Blocking: 1% BSA in PBST for 30 min
4. Primary antibody incubation (anti-vimentin AB28028): in 1% BSA in PBST for overnight (4°C)
5. Secondary antibody incubation (goat anti-mouse ab): in 1% BSA in PBST for 30 min
6. Mounting: Fluoroshield™

Dr. Egi Kardia

Verified customer

Submitted Sep 11 2018

ab4648 staining GFAP in rat brain tissue sections by IHC-P (Formalin/PFA-fixed paraffin-embedded sections)

 Excellent
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
ab4648 staining GFAP in rat brain tissue sections by IHC-P (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with paraformaldehyde, permeabilized with 0.1% Tween 20 in PBS and blocked with 10% Serum for 40 minutes at 25°C. Samples were incubated with primary antibody (1/1500 in TBS) for 24 hours at 4°C. ab96871 was used as the secondary antibody at a dilution of 1/200. ( Green = GFAP immunostained astrocytes, Blue = DAPI stained nuclei.)

Miss. Chia-Li Lin

Verified customer

Submitted Dec 03 2015

ICC on LNCaP cells

 Good
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
1. Cells are Fixed in 2% Paraformaldehyde, (PFA) - 45min at Room Temperature.
2. Permeabilized with 0.5% TritonX -100 in TBS for 7min - 3washes
3. Proteins are blocked with 2% BSA in TBS with 0.1% TritonX -100 for 2 hours.
4. The concentration of DyLight 488 used for this experiment is 1:500 and incubated for 2 hours.

Abcam user community

Verified customer

Submitted Aug 19 2015

Question

1. Alexa Fluor® 488 goat anti-mouse Ig Cat # G A-11001

2. Alexa Fluor® 488 goat anti-rabbit IgG Cat # A- 11008

3. Cy3® goat anti-mouse IgG (H+L) *2mg/mL Cat A 10521

4. Cy3® goat anti-rabbit IgG (H+L) *2mg/mL A10520

5. Cdc2 p34 200 ìg (PSTAIRE sc-53(Epitope is of human origin

6. Nanog Antibody (A-11) sc-374001

7. Oct-3/4 Antibody (H-134) sc-9081

8. Anti Sox2 Cat. No.sc-17320

9. caspase-3 Antibody (H- 277): sc-7148

10. caspase-2L Antibody (C- 19): sc-626

11. Blasticidin S HCl R210-01 50mg)



Please let us have best alternate products for the above at your earliest.


Sincere regards,

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Answer

Thank you for contacting us.

1. The alternative to Alexa Fluor® 488 goat anti-mouse Ig Cat # G A-11001 is ab96871; https://www.abcam.com/Goat-polyclonal-Secondary-Antibody-to-Mouse-IgG-H-L-DyLight-488-ab96871.html

or ab96879; https://www.abcam.com/Goat-polyclonal-Secondary-Antibody-to-Mouse-IgG-H-L-DyLight-488-pre-adsorbed-ab96879.html

or ab150113; https://www.abcam.com/goat-polyclonal-secondary-antibody-to-mouse-igg-hl-alexa-fluorreg-488-ab150113.html

2. ab96895; https://www.abcam.com/Goat-polyclonal-Secondary-Antibody-to-Rabbit-IgG-H-L-DyLight-488-pre-adsorbed-ab96895.html

or ab150077; https://www.abcam.com/goat-polyclonal-secondary-antibody-to-rabbit-igg-hl-alexa-fluorreg-488-ab150077.html

3. ab97035; https://www.abcam.com/Goat-polyclonal-Secondary-Antibody-to-Mouse-IgG-H-L-Cy3-pre-adsorbed-ab97035.html

4. ab97075; https://www.abcam.com/Goat-polyclonal-Secondary-Antibody-to-Rabbit-IgG-H-L-Cy3-pre-adsorbed-ab97075.html

5. ab10345; https://www.abcam.com/PSTAIR-antibody-PSTAIR-ab10345.html

6. We do not have anti Nanog mouse monoclonal that customer is after. We have similar rabbit polyclonal antibodies available. The links are

https://www.abcam.com/Nanog-antibody-ab70482.html

https://www.abcam.com/Nanog-antibody-ab90640.html

7. ab109187; https://www.abcam.com/Oct4-antibody-EPR2053-2-ab109187.html

8. ab97959; https://www.abcam.com/SOX2-antibody-ab97959.html

9. ab131000; https://www.abcam.com/Caspase-3-antibody-ab131000.html

10. ab18737; https://www.abcam.com/Caspase-2-antibody-ab18737.html

11. We do not sell this chemical.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
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Question

Dear xxxxxx,

Below is the reply from my customer:

I wish to use the antibodies in a co-localization experiment. We have primary antibodies against specific Plasmodium falciparum proteins which localize to two different parasite organelles. The fluorescent secondary antibodies will then allow these organelles to be visualized.

We have not performed an experiment like this in the lab before and we will have to develop a protocol. This is the reason why I was wondering what the recommended protocol would be for the DyLight secondary antibodies.

I'm on a very strict budget and I'm looking for the most cost effective way to perform this experiment. In comparison, the Alexa Fluor secondary antibodies have a working concentration of 1ug/ml, with a supplied concentration of 2mg/ml. Do the DyLight secondary antibodies, with a supplied concentration of 0.5mg/ml, have a lower working concentration than that of the Alexa Fluor?

Thank you for your help

Regards

Read More
Answer

Thank you for explaining this for me.

The antibodies mentioned, ab96875,ab96871,ab96877 andab96873 are all recommended to be used in a range of 1/50-1/500 for ICC and IF applications. This is only a starting point and optimisation of these conditions will be required by the end used and their particular experimental conditions. This will be the case with any secondary antibody. This equates to a final concentration of 1 to 10 ug/mL.

How many secondary antibodies are you looking to purchase? It may be that I can arrange for a discount for you?

I hope this information has been of help. I look forward to receiving your reply.

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Question

Dear tech Support

I have another query from a customer. I did send her the data sheets and concentration supplied. On the data sheet the suggested dilution is given, but she still has the following question. Please help. The antibodies in question:
AB96875
AB96871
AB96877
AB96873

Thank you for all your help

Regards

I need to know the concentration of antibody needed per reaction? So what volume of the given stock would I need to use in one experiment?
Best regards

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Answer

Thank you for contacting us and your interest in our products.

The volume of antibody required per experiment would depend on what experiment the customer is hoping to perform, and what volume of diluted antibody the customer usually used in these experiments.

If the customer could please answer the following questions I can help in their request:

1. What application are you hoping to use these antibodies in?

2. Using your protocol, what volume of diluted antibody do you usually apply?

I look forward to receiving your reply.

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Question

I have used ab10062 and ab7751 to stain dorsal root ganglion via ICC and mouse tissue extract via WB; there was no signal from either experiments, for ICC i used 1:100 dilution of primary, fixed with 4% PFA, perm with triton x-100; blocked with milk and incubated primary overnight at 4 deg C; used ab96871 as secondary;

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Answer

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement for one vial of ab98787.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

Read More

Question

Dear All,



Sending you attached a list of the tendered items. You can see the products at page 3-5. Please give all of the items a check and let us know which items are best available with Abcam.



Early response requested.



Sincere regards,

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Answer

Thank you for your inquiry.

We have a number of products that are comparable with the products on the list you sent us. For future inquiriesI would like to direct you to the search facility on our website (www.abcam.com).

Please enter the name of the protein into the search box at the top of the home page. A list of products will be generated which can be further refined using the filters on the left hand side of the page. Using these filters will help you to identify products that have been tested in the applications and species you are interested in.

Please find a list of the product code and the links to the respective datasheets below:

ab1012 (Anti-Histone H3 (tri methyl K4) antibody [mAbcam1012] - ChIP Grade)

https://www.abcam.com/index.html?datasheet=1012 (or use the following: https://www.abcam.com/index.html?datasheet=1012).

ab12209 (Anti-Histone H3 (tri methyl K4) antibody [mAbcam12209] - ChIP Grade)

https://www.abcam.com/index.html?datasheet=12209 (or use the following: https://www.abcam.com/index.html?datasheet=12209).

ab6002 (Anti-Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade)

https://www.abcam.com/index.html?datasheet=6002 (or use the following: https://www.abcam.com/index.html?datasheet=6002).

ab124462 (Anti-Flag Tag antibody)

https://www.abcam.com/index.html?datasheet=124462 (or use the following: https://www.abcam.com/index.html?datasheet=124462).

ab6939 (Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Cy3 ®), pre-adsorbed)

https://www.abcam.com/index.html?datasheet=6939 (or use the following: https://www.abcam.com/index.html?datasheet=6939).

ab96899 (Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (DyLight® 488), pre-adsorbed)

https://www.abcam.com/index.html?datasheet=96899 (or use the following: https://www.abcam.com/index.html?datasheet=96899).

ab96919 (Donkey anti-Rabbit IgG H&L (DyLight® 488) secondary antibody)

https://www.abcam.com/index.html?datasheet=96919 (or use the following: https://www.abcam.com/index.html?datasheet=96919).

ab96879 (Goat polyclonal Secondary Antibody to Mouse IgG - H&L (DyLight® 488), pre-adsorbed)

https://www.abcam.com/index.html?datasheet=96879 (or use the following: https://www.abcam.com/index.html?datasheet=96879).

ab96871 (Goat anti-Mouse IgG H&L (DyLight® 488) secondary antibody)

https://www.abcam.com/index.html?datasheet=96871 (or use the following: https://www.abcam.com/index.html?datasheet=96871).

ab97265 (Goat anti-Mouse IgG Fc (HRP) secondary antibody)

https://www.abcam.com/index.html?datasheet=97265 (or use the following: https://www.abcam.com/index.html?datasheet=97265).

ab7481 (Goat Serum)

https://www.abcam.com/index.html?datasheet=7481 (or use the following: https://www.abcam.com/index.html?datasheet=7481).

ab7487 (Rabbit Serum (sterile))

https://www.abcam.com/index.html?datasheet=7487 (or use the following: https://www.abcam.com/index.html?datasheet=7487).

ab46666 (Herring sperm ssDNA)

https://www.abcam.com/index.html?datasheet=46666 (or use the following: https://www.abcam.com/index.html?datasheet=46666).

ab52579 (Ribonuclease A protein (Active))

https://www.abcam.com/index.html?datasheet=52579 (or use the following: https://www.abcam.com/index.html?datasheet=52579).

ab73430 (DNase I protein (Active))

https://www.abcam.com/index.html?datasheet=73430 (or use the following: https://www.abcam.com/index.html?datasheet=73430).

ab64220 (Proteinase K)

https://www.abcam.com/index.html?datasheet=64220 (or use the following: https://www.abcam.com/index.html?datasheet=64220).

ab65621 (Protease Inhibitor Cocktail)

https://www.abcam.com/index.html?datasheet=65621 (or use the following: https://www.abcam.com/index.html?datasheet=65621).

ab116028 (Prism Ultra Protein Ladder (10-245 kDa))

https://www.abcam.com/index.html?datasheet=116028 (or use the following: https://www.abcam.com/index.html?datasheet=116028).


I hope this information is helpful to you. If you have specific questions about the products identified during the search, please do not hesitate to contact me.

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Question

This customer has purchased ab96885 (Goat anti-Rabbit IgG H&L (DyLight® 594) secondary antibody), ab96871 (Goat anti-Mouse IgG H&L (DyLight® 488) secondary antibody) and has conducted the ICC several times. This customer indicated she will observe the PE fluorescence when she wants to observe the FITC fluorescence, but this state doesn’t happen when she wants to observe the PE fluorescence, therefore, she would like to ask for your help to find out any possible problem and also help her to modify her experiment step, could you please offer any suggestion to her?

I attached the image in this letter and her experiment step as follow:





1) Order details:

˙Batch number (Lot number): ab96885--gr46891-6 and ab96871--gr50260-4

Po: 1028918

˙Abcam product code: ab96885, ab96871

˙Antibody storage conditions (temperature/reconstitution etc) : 4℃



2) Please describe the problem (high background, non-specific signal…etc).

I can observe the PE fluorescence when I want to observe the FITC fluorescence

3) On what material are you testing the antibody in IHC/ICC?

˙Species: 1% BSA, in 1X PBS

˙What’s cell line or tissue: sclera cell



4) Sample preparation:

˙4%PFA/formalin fixed

˙cells in culture

˙Sample preparation:

˙Positive control :

˙Negative control :



5) Fixation step

˙Yes or No : Yes

˙If yes, Fixative agent and concentration: 4%PFA/formalin fixed

˙Fixation time: 5min

˙Fixation temperature :R.T.



6) Antigen retrieval method(including time, temperature etc.):

R.T. 60min



7) Permeabilization method:

˙Did you do a permeabilization step (details please) or add permeabilizing agent in any dilution buffers?

˙Permeabilizing agent and concentration:





8) Blocking agent (eg BSA, serum…):

˙Concentration:1% BSA

˙Blocking time : 60min

˙Blocking temperature: R.T.



9)

˙Endogenous peroxidases blocked?

˙Endogenous biotins blocked?



10) Primary antibody (If more than one was used, describe in “additional notes”) :

˙Species:

˙Reacts against:

˙At what dilution(s) have you tested this antibody:1:50˜250

˙Diluent buffer: 1% BSA in 1xPBS

˙Incubation time: 60min

˙Incubation temperature: R.T.

˙What washing steps were done (which buffer, number of washes): 3times, 5min



11) Secondary antibody:

˙Species:

˙Reacts against which species:

˙At what dilution(s) have you tested this antibody: 1:1000

˙Diluent buffer: 1% BSA in 1xPBS

˙Incubation time: 60min

˙What washing steps were done (which buffer, number of washes): 3times, 5min

˙Fluorochrome or enzyme conjugate (eg: FITC, HRP, AP, biotin…etc): FITC and TRITC

˙Do you know whether the problems you are experiencing come from the secondary?



12) Signal amplification method (eg: ABC, LSAB, HRP polymer, TSE):



13) Detection method (eg: DAB, BCIP/NBT …etc):



14)

˙ How many times have you run this staining? 2

˙Do you obtain the same results every time? YES

˙What steps have you altered to try and optimize the use of this antibody?





Could you please help this customer to solve the problem?

Thanks for your kindly help

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Answer

Thank you for taking time to complete our questionnaire. I am sorry to hear that this antibody is not providing satisfactory results.

The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality.

Having reviewed this case, I would like to offer some suggestions to help optimize the results from abab96885 Goat anti-Rabbit IgG H&L (DyLight® 594) secondary antibody. I would also appreciate if you can confirm some further details:

1. What kind of filter or lamp are you using? Unfortunately, DyLight® 488and DyLight® 594 do overlap: a little bit in the DyLight® 488 spectrum, but more important in the DyLight® 594 wavelengths, which corresponce with your findings. Check with the manufacturer of your microscope if their filters are suitable for those dyes.

2. In order to check if the ab96885 is damaged I would suggest to perform an ICC experimentwith only one primary- secondaryreaction. This way you canexclude the annoying "bleed-over".


Should the suggestions not improve the results, please do let me know.

In the event that a product is not functioning in the species and applications cited on the product datasheet (and the problem has been reported within 6 months of purchase), we would be pleased to provide a free of charge replacement, credit note, or refund.

I hope this information is helpful, and I thank you for your cooperation.

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Question

Dear Sir/ Madam,

I wanna to study the N-cadherin for A7R5 (one kind of Mouse smooth muscle cell) and currentlyI have ordered the ab98952 but for the online sheet there is no 'secondary antibodies' tab on this product. May I know what kind of secondary antibody with FITC could be suitable for ab98952, for example, isGoat anti-Mouse IgG1 heavy chain (FITC) secondary antibody (a b97239) suitable? And I also wanna to know is there any conjugated antibody (conjugated withfluorescent secondary antibody) for N-cadherin? How about productAnti-N Cadherin antibody [GC-4], please recommend some for me, thanks a lot.

I am looking forward to your reply, and I hope I can order the right product ASAP.

Thanks a lot.

Best wishes,

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Answer

Thank you for your enquiry and interest.

You could certainly use our advanced search engine specifically designed for secondary antibodies to find the most suitable products for your studies:

https://www.abcam.com/index.html?pageconfig=productmap&cl=918



Since the isotype of this primary antibody is mouse IgG1, so you could select the isotype, target species, host species, format, specificity, conjugate using the drop-down menu.

We have different conjugates such as FITC, DyLight488, Chromeo 488, and here are a few suggestions:

FITC:

ab98692

ab97239

ab11588

You can find some further information about the the generation of our fluorochromes at this site:https://www.abcam.com/index.html?pageconfig=resource&rid=12948

DyLight488:

Affinity purified: ab96875, ab97015, ab96871

Pre-adsorbed: ab96879

Chromeo 488:

ab60313



If you need any further assistance in the future, please do not hesitate to contact me.

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1-10 of 12 Abreviews or Q&A

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