Overview

  • Product name
    Goat Anti-Mouse IgG H&L (HRP)
    See all IgG secondary antibodies
  • 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 (ab182017): Chicken IgY, less than 2%. Human IgG, less than 6%. Rabbit IgG, less than 7%. Rat IgG, less than 47%.
  • Tested applications
    Suitable for: WB, IP, ELISA, IHC-Pmore details
  • 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: 7.4
    Preservative: 0.1% Proclin
    Constituents: PBS, 1% BSA, 30% Glycerol
  • 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

Applications

Our Abpromise guarantee covers the use of ab205719 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 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

  • All lanes : Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) 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
    Goat Anti-Mouse IgG H&L (HRP) (ab205719) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Observed band size : 52 kDa (why is the actual band size different from the predicted?)


    Exposure time : 5 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 ab7291 overnight at 4°C. Antibody binding was detected using ab205719, and visualised using ECL development solution ab133406.

  • IHC image of alpha tubulin staining in a section of formalin-fixed paraffin-embedded normal human colon tissue*. 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 1/1000 dilution. An HRP-conjugated secondary (Ab205719, 1/10000 dilution) was used to detect the primary 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

  • All lanes : Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) 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
    ab205719 (Left Image) at 1/5000 and a competitor secondary (Right Image) at 1/5000. Notice the decreased signal of the competitor product.

    Performed under reducing conditions.

    Observed band size : 52 kDa (why is the actual band size different from the predicted?)


    Exposure time : 5 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 ab7291 overnight at 4°C. Antibody binding was detected using ab205719 (Left Image) and a competitor secondary (Right Image), and visualised using ECL development solution ab133406.

  • 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
    Goat Anti-Mouse IgG H&L (HRP) (ab205719) at 1/2000 dilution

    Performed under reducing conditions.

    Exposure time : 10 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 (ab205719), and visualised using ECL development solution ab133406.

  • IHC image of histone H4 staining in a section of formalin-fixed paraffin-embedded normal human colon tissue*. 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 1/1000 dilution. An HRP-conjugated secondary (Ab205719, 1/10000 dilution) was used to detect the primary 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

  • 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
    ab205719 (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 : 10 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 ab205719 (Left Image) and a competitor secondary (Right Image), and visualised using ECL development solution ab133406.

  • 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
    Goat Anti-Mouse IgG H&L (HRP) (ab205719) at 1/5000 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 : 10 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 ab205719, and visualised using ECL development solution ab133406.

References

This product has been referenced in:
  • Li Z  et al. Hyperbaric oxygen inhibits venous neointimal hyperplasia following arteriovenous fistulization. Int J Mol Med 39:1299-1306 (2017). Read more (PubMed: 28393184) »
  • Kong D  et al. Acetamiprid inhibits testosterone synthesis by affecting the mitochondrial function and cytoplasmic adenosine triphosphate production in rat Leydig cells. Biol Reprod 96:254-265 (2017). Read more (PubMed: 28395332) »

See all 7 Publications for this product

Customer reviews and Q&As

Application
Western blot
- 20ug of bovine liver lysates were loaded on to 4-15% gradient gel
- Blocked for an hour in 5% skim milk
- 1:4000 primary antibody used (ab110413). Incubation O/N @4C
- 1:20000 secondary antibody used (ab205719). Incubation 1 Hour room temperature.
Username

Julie Kim

Verified customer

Submitted Apr 21 2017

Abreviews
Application
Western blot
Whole Cell lysate - human ES cells.
Loading amount: 10 µg.
Gel Running Conditions: Reduced Denaturing (10%).
Blocking step: 5%BSA/1%OvaAlb/PBS as blocking agent for 3 hours at room temperature.
Primary antibody: Nanog diluted 1:1000 in 1%BSA/0.2%OvaAlb/PBS 13hours at 4°C.
Secondary antibody: ab205719 - Goat Anti-Mouse IgG H&L (HRP) diluted 1:5000 in TBST.
ECL prime: 4 second exposure.
Predicted size: ~45kDa.
Username

Abcam user community

Verified customer

Submitted Aug 12 2016

Application
Western blot
Excellent specificity.
Excellent sensitivity.
WB conditions:
- Cell lisates: 20ug 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
- Exposure time: 1 second, 3th film out 4
Username

Abcam user community

Verified customer

Submitted Feb 01 2016

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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