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  1. Link

    products/secondary-antibodies/donkey-mouse-igg-hl-hrp-ab205724.pdf

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Immunology Immunoglobulins Heavy Chain IgG
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Donkey Anti-Mouse IgG H&L (HRP) (ab205724)

  • Datasheet
  • SDS
Reviews (2) Submit a question References (2)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Donkey Anti-Mouse IgG H&L (HRP) (ab205724)
  • Western blot - Donkey Anti-Mouse IgG H&L (HRP) (ab205724)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Donkey Anti-Mouse IgG H&L (HRP) (ab205724)
  • Western blot - Donkey Anti-Mouse IgG H&L (HRP) (ab205724)
  • Western blot - Donkey Anti-Mouse IgG H&L (HRP) (ab205724)
  • Western blot - Donkey Anti-Mouse IgG H&L (HRP) (ab205724)

Key features and details

  • Donkey Anti-Mouse IgG H&L (HRP)
  • Conjugation: HRP
  • Host species: Donkey
  • Isotype: IgG
  • Suitable for: WB, IP, ELISA, IHC-P

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Overview

  • Product name

    Donkey Anti-Mouse IgG H&L (HRP)
    See all IgG secondary antibodies
  • Host species

    Donkey
  • Target species

    Mouse
  • Specificity

    The antibody used for conjugation reacts with mouse immunoglobulins of all classes. Cross-reactions as determined by ELISA for the unconjugated antibody (ab182022): Human IgG, rabbit IgG, goat IgG and Chicken IgY, less than 2%. Rat IgG, less than 33%.
  • Tested applications

    Suitable for: WB, IP, ELISA, IHC-Pmore details
  • Immunogen

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

  • Conjugation

    HRP

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. Store In the Dark.
  • Storage buffer

    pH: 7.40
    Preservative: 0.1% Proclin 300 Solution
    Constituents: PBS, 1% BSA, 30% Glycerol (glycerin, glycerine)
  • 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 Horse Radish Peroxidase (HRP).
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Immunology
    • Immunoglobulins
    • Heavy Chain
    • IgG
    • Secondary antibodies
    • anti-Mouse
    • IgG
    • Enzyme
    • HRP

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab205724 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
WB (2)
1/2000 - 1/20000.
IP
Use at an assay dependent concentration.
ELISA
Use at an assay dependent concentration.
IHC-P
1/2000 - 1/20000.
Notes
WB
1/2000 - 1/20000.
IP
Use at an assay dependent concentration.
ELISA
Use at an assay dependent concentration.
IHC-P
1/2000 - 1/20000.

Images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Donkey Anti-Mouse IgG H&L (HRP) (ab205724)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Donkey Anti-Mouse IgG H&L (HRP) (ab205724)

    IHC image of Histone H4 staining in a section of formalin-fixed paraffin-embedded normal human colon*. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6) for 30mins, and incubated overnight at +4°C with ab31830 at 0.5ug/ml. DAB was used as the chromogen (ab103723), diluted 1/100 and incubated for 10min at room temperature.

    An HRP-conjugated secondary (Ab205724, 1/2000 dilution) was used for 1hr at room temperature.

    The section was counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • Western blot - Donkey Anti-Mouse IgG H&L (HRP) (ab205724)
    Western blot - Donkey Anti-Mouse IgG H&L (HRP) (ab205724)
    All lanes : Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1 µg/ml

    Lane 1 : Liver (Human) Tissue Lysate
    Lane 2 : Liver (Mouse) Tissue Lysate
    Lane 3 : Liver (Rat) Tissue Lysate
    Lane 4 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 5 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 6 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Donkey Anti-Mouse IgG H&L (HRP) (ab205724) at 1/2000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Observed band size: 42 kDa why is the actual band size different from the predicted?


    Exposure time: 15 seconds


    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab8226 overnight at 4°C. Antibody binding was detected using ab205724, and visualised using ECL development solution ab133406.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Donkey Anti-Mouse IgG H&L (HRP) (ab205724)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Donkey Anti-Mouse IgG H&L (HRP) (ab205724)

    IHC image of alpha tubulin staining in a section of formalin-fixed paraffin-embedded normal human colon*. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6) for 30mins, and incubated overnight at +4°C with ab7291 at 0.5ug/ml.

    An HRP-conjugated secondary (Ab205725, 1/2000 dilution) was used for 1hr at room temperature. DAB was used as the chromogen (ab103723), diluted 1/100 and incubated for 10min at room temperature. The section was counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • Western blot - Donkey Anti-Mouse IgG H&L (HRP) (ab205724)
    Western blot - Donkey Anti-Mouse IgG H&L (HRP) (ab205724)
    All lanes : No Primary Antibody

    Lane 1 : Liver (Human) Tissue Lysate
    Lane 2 : Liver (Mouse) Tissue Lysate
    Lane 3 : Liver (Rat) Tissue Lysate
    Lane 4 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 5 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 6 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Donkey Anti-Mouse IgG H&L (HRP) (ab205724) at 1/2000 dilution

    Performed under reducing conditions.

    Exposure time: 15 seconds


    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was incubated overnight with 2% Bovine Serum Albumin at 4°C. Any non-specific background binding was assessed by incubating the membrane with only the secondary antibody (ab205724), and visualised using ECL development solution ab133406.

  • Western blot - Donkey Anti-Mouse IgG H&L (HRP) (ab205724)
    Western blot - Donkey Anti-Mouse IgG H&L (HRP) (ab205724)
    All lanes : Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1 µg/ml

    Lane 1 : Liver (Human) Tissue Lysate
    Lane 2 : Liver (Mouse) Tissue Lysate
    Lane 3 : Liver (Rat) Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : ab205724 (Left Image) at 1/2000 and a competitor secondary (Right Image) at 1/2000. Notice the increased background of the competitor product.

    Performed under reducing conditions.

    Observed band size: 42 kDa why is the actual band size different from the predicted?


    Exposure time: 15 seconds


    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab8226 overnight at 4°C. Antibody binding was detected using ab205724 (Left Image) and a competitor secondary (Right Image), and visualised using ECL development solution ab133406.

  • Western blot - Donkey Anti-Mouse IgG H&L (HRP) (ab205724)
    Western blot - Donkey Anti-Mouse IgG H&L (HRP) (ab205724)
    All lanes : No Primary Antibody

    Lane 1 : Liver (Human) Tissue Lysate
    Lane 2 : Liver (Mouse) Tissue Lysate
    Lane 3 : Liver (Rat) Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : ab205724 (Left Image) 1/2000 and a competitor secondary (Right Image) 1/2000. Notice the increased background of the competitor product.

    Performed under reducing conditions.

    Exposure time: 15 seconds


    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was incubated overnight with 2% Bovine Serum Albumin at 4°C. Any non-specific background binding was assessed by incubating the membrane with ab205724 (Left Image) and a competitor secondary (Right Image), and visualised using ECL development solution ab133406.

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 (2)

Publishing research using ab205724? Please let us know so that we can cite the reference in this datasheet.

ab205724 has been referenced in 2 publications.

  • Dorandish S  et al. Regulation of amyloid-ß levels by matrix metalloproteinase-2/9 (MMP2/9) in the media of lung cancer cells. Sci Rep 11:9708 (2021). PubMed: 33958632
  • Maggi EC  et al. Retinoblastoma-binding protein 2 (RBP2) is frequently expressed in neuroendocrine tumors and promotes the neoplastic phenotype. Oncogenesis 5:e257 (2016). PubMed: 27548814

Customer reviews and Q&As

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

1-2 of 2 Abreviews or Q&A

GAPDH western

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
5ug liver lysate (abcam ab29889) 10ug whole cell lysate (human ES cells)
Blocking buffer - 5%BSA/1%OvAlb/PBS (also works with 5% milk powder) for 3hours
Primary antibody - GAPDH (1:10000 expected size 37kDa) 14hours at 4C
Secondary diluted in TBST (1:5000) for 3 hours at room temperature 3hours at room temperature
ECL prime
5 second exposure
Lane 1. Liver lysate
Lane 2. Ladder
Lane 3. Human ES cell lysate
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 24 2016

Test ab205724 - Donkey Anti-Mouse IgG H&L (HRP)

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Excellent specificity.
Good sensitivity.
WB conditions:
- Cell lisates: 20 ug protein
- non-reducing
- Primary antibody: Mouse anti-GRP78, clon 40/BiP, Cat. No. 610979. BD Biosciences. Dilution 1:1000
- Secondary antibody: 1:6000
- Developer: ECL Prime, Amersham, GE Helathcare Life Sciences, Product code RPN2232
- Expusure time: 1 second, 3th film out 4
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

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