Overview

  • Product name
    Donkey Anti-Goat IgG H&L (HRP) preadsorbed
    See all IgG secondary antibodies
  • Description
    Donkey polyclonal Secondary Antibody to Goat IgG - H&L (HRP), pre-adsorbed
  • Host species
    Donkey
  • Target species
    Goat
  • Specificity

    By immunoelectrophoresis and ELISA this antibody reacts specifically with Goat IgG and with light chains common to other Goat immunoglobulins. No antibody was detected against non immunoglobulin serum proteins. Reduced cross-reactivity to chicken, horse, human, mouse, pig, rabbit and rat was detected.

     

  • Tested applications
    Suitable for: ICC, IHC-P, ELISA, WBmore details
  • Minimal
    cross-reactivity

    Chicken, Human, Mouse, Pig, Rabbit, Rat more details
  • Conjugation
    HRP

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C.
  • Storage buffer
    Preservative: 0.1% Proclin
    Constituents: 0.2% BSA, PBS
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Purification notes
    Antiserum was cross adsorbed using chicken, human, mouse, pig, rabbit and rat immunosorbents to remove cross reactive antibodies. This antibody was isolated by affinity chromatography using antigen coupled to agarose beads and conjugated to Horse Radish Peroxidase (HRP).
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab97120 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 Use at an assay dependent dilution.
IHC-P 1/200 - 1/5000.
ELISA 1/10000 - 1/100000. (Primary)
WB 1/5000 - 1/50000. Colorimetric 1/5000 - 1/30000; Chemiluminescent 1/10000 - 1/50000

References

This product has been referenced in:
  • Dominy SS  et al. Porphyromonas gingivalis in Alzheimer's disease brains: Evidence for disease causation and treatment with small-molecule inhibitors. Sci Adv 5:eaau3333 (2019). Read more (PubMed: 30746447) »
  • Pittala S  et al. Targeting Liver Cancer and Associated Pathologies in Mice with a Mitochondrial VDAC1-Based Peptide. Neoplasia 20:594-609 (2018). IHC-P . Read more (PubMed: 29747160) »
See all 10 Publications for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Answer



I am happy to confirm that we have a anti goat antibody that was pre-absorped against sheep IgG:

ab99713

https://www.abcam.com/index.html?datasheet=99713 (or use the following: https://www.abcam.com/index.html?datasheet=99713).

Please review the datasheet for more information.

This antibody is tested and guaranteed for ELISA as well.

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Question
Answer

Thank you for contacting Abcam.


Attached please find the CoC document you requested.


I hope this information is helpful. Please do not hesitate to contact us if you have any additional questions.

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Question

Dear Tech Support Team, Please see below the details of customer's complaint. Attached is the image of the gel. Please advise. Antibody code: Ab9134 Batch number: Gr4307-5 Antibody storage conditions (temperature/reconstitution etc) 40C Description of the problem (high background, wrong band size, more bands, no band etc.) We got a strong band of 90kDa and weak band of the protein with the HA- tag and in the negative control (lymphocytes) we got just the wrong band. We also checked the secondary antibody against the SDS PAGE without the first antibody and the blot was clean. Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.) Lymphocytes cell extract, nuclear and cytoplasmatic extract of HEK 293 cells with the HA tag over-expression   Sample preparation (Buffer/Protease inhibitors/Heating sample etc.) The cells lysed in RIPA buffer of 50 mM Tris pH 8.0, 150 mM NaCl, 5 mM EDTA pH 8.0, 1% NP-40, 0.5% Sodium deoxycholate (DOC), 0.1% SDS and incubated on ice for 30 min. Loading buffer (10% SDS , 10mM beta-mercapto-ethanol,20 %  Glycerol, 0.2 M Tris-HCl, pH 6.8, 0.05% Bromophenolblue) added to the samples and they boiled for 10 minutes and loaded on the gel.   Amount of protein loaded 167000 cells per well Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.) The samples run on the gel of 4-15% Mini protean TGX –BioRAD. Transfer and blocking conditions (Buffer/time period, Blocking agent etc.) The gel transferred for 90 min. on 150 mA in transfer buffer (25 mM Tris-HCl, 192 mM  Glycine, 0.1% SDS, 20% Methanol) and blocked in 1% skim milk with 0.3% Tween-20 in PBS, for an hour at room temperature. Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step) The antibody diluted 1:1000 in 1% skim milk with 0.3% Tween-20 in PBS, over night at 40C.  The membrane washed three times with PBS with 0.3% Tween-20.  Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step) The secondary antibody donkey polyclonal to Goat IgG (ab97120) diluted 1:12000 in 1% skim milk with 0.3% Tween-20 in PBS and incubated at room temperature for an hour. The membrane washed three times with PBS with 0.3% Tween-20.    Detection method (ECL, ECLPlus etc.) SuperSignal westpico chemiluminescent substrate (Pierce) used as a detection. Positive and negative controls used (please specify) We used the lymphocytes as a negative control but we got the wrong band on them also. We didn’t use any positive control.   Optimization attempts (problem solving)   How many times have you tried the Western? Twice Have you run a "No Primary" control? No we didn’t use that control but we used the same blot with the primary antibody of ab96326. Do you obtain the same results every time? e.g. are the background bands always in the same place? Yes What steps have you altered? We used different over-expression with the HA-tag.  

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Answer

Thank you for contacting us. I have read the whole protocol you have kindly submitted. In order to address the problem quickly I have further questions to ask. - According to the attached image you have used non transfected N2a mouse cells, Lymphocytes supernatant and Lymphocytes pellet as a negative control right? I am sorry I am surprised by the fact that 1st and 3rd (very very faint band may due to spillage from 4th lane) there is no band while there is dark band in 2nd lane. Could you confirm there was no intermixing of lysates? - What is the band size you are expecting to be? - The description of the problem also does not correlate with the image. Does this mean the Lymphocyte pellet has HA-tag inserted cells? I predict from the image; the problem only is that antibody is detecting a band in Lymphocyte supernatant which it should not do. Right? - Could you specify which protein you have tagged with HA? Have you done any western blot studies with anti protein antibody? - Could you provide the order details so that I check if the delivery was on time? I will look forward to hearing from you soon.

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Answer

Thank you for contacting Abcam.   I have attached the MSDS for ab97120 and ab97105, both of which contain the preservative proclin.  A MSDS is not required for ab105466.   I hope this information is helpful.  Please do not hesitate to contact us if you have any additional questions.

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