Goat Anti-Mouse IgG H&L (FITC) preadsorbed (ab7064)

Reviews (3)Q&A (2)References (8)

Overview

  • Product name
    Goat Anti-Mouse IgG H&L (FITC) preadsorbed
    See all IgG secondary antibodies
  • Host species
    Goat
  • Target species
    Mouse
  • Tested applications
    Suitable for: ICC/IF, Immunomicroscopy, Flow Cyt, IHC-P, IHC-Fr, ELISAmore details
  • Minimal
    cross-reactivity

    Chicken, Cow, Goat, Guinea pig, Hamster, Horse, Human, Rabbit, Rat, Sheep more details
  • Immunogen

    Full length native Mouse IgG (purified).

  • Conjugation
    FITC. Ex: 493nm, Em: 528nm

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C.
  • Storage buffer
    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 0.42% Potassium phosphate, 0.87% Sodium chloride, 1% BSA
  • Concentration information loading...
  • Purity
    Affinity purified
  • Purification notes
    This product was prepared from monospecific antiserum by immunoaffinity chromatography using mouse IgG coupled to agarose beads followed by solid phase adsorption(s) to remove any unwanted reactivities.
  • Conjugation notes
    Diaminotriazinylaminofluorescein (DTAF) (Molecular Weight 530 daltons) Absorption Wavelength: 495 nm Emission Wavelength: 528 nm Fluorochrome/Protein Ratio: 2.3 moles DTAF per mole of Rabbit IgG
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Associated products

Applications

Our Abpromise guarantee covers the use of ab7064 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 Use at an assay dependent concentration. PubMed: 15767251
Immunomicroscopy Use at an assay dependent concentration.
Flow Cyt 1/1000 - 1/2000.
IHC-P Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
ELISA 1/10000 - 1/50000.

Images

  • Immunocytochemistry/ Immunofluorescence - Goat Anti-Mouse IgG H&L (FITC) preadsorbed (ab7064)
    Immunocytochemistry/ Immunofluorescence - Goat Anti-Mouse IgG H&L (FITC) preadsorbed (ab7064)Humphries, W.H. 4th et al PLoS One. 2011;6(10):e26626. doi: 10.1371/journal.pone.0026626. Epub 2011 Oct 24

    Rab7 and LAMP1 are highly colocalized in BS-C-1 cells and in HeLa cells

    LAMP1  was detected with a mouse antiLAMP1 antibody in ICC/IF analysis of BS-C-1 cells.

    Anti-LAMP1 was visualized using ab7064 at 1/500 diulution.

  • Flow Cytometry - Goat Anti-Mouse IgG H&L (FITC) preadsorbed (ab7064)
    Flow Cytometry - Goat Anti-Mouse IgG H&L (FITC) preadsorbed (ab7064)
    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 (FITC, pre-adsorbed) (ab7064) was used at 1/1000 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.
  • Immunocytochemistry/ Immunofluorescence - Goat polyclonal Secondary Antibody to Mouse IgG - H&L (FITC), pre-adsorbed (ab7064)
    Immunocytochemistry/ Immunofluorescence - Goat polyclonal Secondary Antibody to Mouse IgG - H&L (FITC), pre-adsorbed (ab7064)
    Ab7064 was used at dilution 1/20 with the primary antibody ab7852 in ICC. See the review on ab7852.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat polyclonal Secondary Antibody to Mouse IgG - H&L (FITC), pre-adsorbed (ab7064)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat polyclonal Secondary Antibody to Mouse IgG - H&L (FITC), pre-adsorbed (ab7064)
    Ab7064 was used at dilution 1/2000 with the primary antibody ab2378 in IHC-P. See the review on ab2378.

References

This product has been referenced in:
  • Ghezzi CE  et al. 3D Functional Corneal Stromal Tissue Equivalent Based on Corneal Stromal Stem Cells and Multi-Layered Silk Film Architecture. PLoS One 12:e0169504 (2017). Read more (PubMed: 28099503) »
  • Liu X  et al. Laminarin protects against hydrogen peroxide-induced oxidative damage in MRC-5 cells possibly via regulating NRF2. PeerJ 5:e3642 (2017). Read more (PubMed: 28785522) »
See all 8 Publications for this product

Publishing research using ab7064? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-5 of 5 Abreviews or Q&A

nice working antibody

 Good
Abreviews
Application
Immunohistochemistry (Frozen sections)
This is a good antibody and a stable conjugate.
The binding is specific.
The luminous intensity of the conjugate is good and is stable (can still be evaluated after a long time).

Abcam user community

Verified customer

Submitted Oct 07 2015

good secendary antibody

 Good
Abreviews
Application
Flow Cytometry
This is a good functioning antibody with a stable conjugate.
The binding is very specific and the luminous intensity of the conjugate is good.
We have used it in the flow cytometry with a 1/1000 dilution

Abcam user community

Verified customer

Submitted Oct 06 2015

Talin Staining

 Good
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Mouse Osteoblast cells were grown on cover slips for 48 hrs. Then Cells were fixed with formaldehyde vapor for 10 mins. Cells were permeabilized with Triton-X 100 for 5-7mins. 3% BSA was used for blocking (1hr). 1:100 dilution of mouse anti-talin antibody to mouse was added onto the cells and incubated for 1hr. Then 1:500 dilution of anti-mouse FITC conjugated secondary antibody was added on to the cells and incubated for 1hr. Before proceeding to each step cells were washed with PBS.

Abcam user community

Verified customer

Submitted Jul 05 2013

Question

Thanks for your kindly reply, this customer already tried to used the other secondary antibody ab6785 (Goat polyclonal Secondary Antibody to Mouse IgG - H&L (FITC)) to conducted the same experiment, but the result still shows weak signal, therefore, she think these antibodies was lost its function already, I also attached the results in this letter, please check it and could you please help this customer to create two credit notes? I am really appreciate for your kindly help : )

Best regards

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Answer

Thank you for confirming this details.

In my understanding this results show that the ab24590 isfaulty as it has been used with two different secondary antibodies without success. I provide you here therefore with the credit note for this antibody. The credit note ID is xxfor the ab24590.

In regards to the ab7064, I expect this antibody to work with another primary mouse antibody.I do not think that this antibody also should be faulty. Pleasedo let me knowhowever if you have anyconcerns in this regards.

Thank you for your help and cooperation also in this case.Please let me know if you have any questions.

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Question

xx product code: ab24590, ab7064 ˙Antibody storage conditions (temperature/reconstitution etc) : -20℃ 2) Please describe the problem (high background, non-specific signal…etc). No staining 3) On what material are you testing the antibody in IHC/ICC? ˙Species: Human ˙What’s cell line or tissue: HUVEC (cell line, P4) 4) Sample preparation: ˙Type of sample (Fresh frozen sections, perfusion fixed frozen sections, PFA/formalin fixed ˙paraffin embedded sections, cells in culture, other:____) : cells in culture with 2% gelatin coating ˙Sample preparation: ˙Positive control : ˙Negative control : 5) Fixation step ˙Yes or No : Yes and no, both way are tried ˙If yes, Fixative agent and concentration: 4% PFA ˙Fixation time: 10min ˙Fixation temperature : room temperature 6) Antigen retrieval method(including time, temperature etc.): no ˙Did you do a permeabilization step (details please) or add permeabilizing agent in any dilution buffers? Yes if fixation is done and no if not fixed ˙Permeabilizing agent and concentration: 0.2% Tween20 after fixation 8) Blocking agent (eg BSA, serum…): ˙Concentration:1%BSA/PBS ˙Blocking time : 30min ˙Blocking temperature: room temperature 9) ˙Endogenous peroxidases blocked? no ˙Endogenous biotins blocked? no 10) Primary antibody (If more than one was used, describe in “additional notes”) : ˙Species: mouse ˙Reacts against: human ˙At what dilution(s) have you tested this antibody: 1:50 and 1:100 ˙Diluent buffer: 1%BSA/PBS ˙Incubation time: 1 hr and 2hr at room temperature or 4°C overnight ˙What washing steps were done (which buffer, number of washes): PBS X3 5min 11) Secondary antibody: gout anti mouse IgG(H+L) (ab7064) ˙Species: goat ˙Reacts against which species: mouse ˙At what dilution(s) have you tested this antibody: 1:20 1:50 and 1:100 ˙Diluent buffer: 1%BSA/PBS ˙Incubation time: 1hr at room temperature ˙What washing steps were done (which buffer, number of washes): PBS X3 5min ˙Fluorochrome or enzyme conjugate (eg: FITC, HRP, AP, biotin…etc): FITC ˙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): Slides were observed under a fluorescence microscope and confocal microscope 14) ˙ How many times have you run this staining? 5 times ˙Do you obtain the same results every time? yes ˙What steps have you altered to try and optimize the use of this antibody? Dilutions of the primary antibody and secondary antibody Incubation time of primary antibody Fixation or no fixation

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Answer

Thank you for taking time to complete our questionnaire and for contacting us. I am sorry to hearthat the customer is notobtaining a staining with this two antibodies.

The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. I appreciate the time the customer has spent in the laboratory. It is regrettable the results have not been successful.

Having reviewed the protocol details, I believe these two products should have given satisfactory results. It appears that you may have received a faulty vial. I apologize for the inconvenience and am pleased to offer you a free of charge replacement, credit note, or refund in compensation.In order however to replace the right antibody, we need toknow whether the primary or the secondary antibody is faulty. Can the customer therefore either test the secondary antibody with another mouse primary antibody which is known to work, or alternatively use another secondary antibody witht hte ab24590 to see whether the problem is due to the secondary antibody?

Thank you for your cooperation. I look forward to hearing from you.

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