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  1. Link

    donkey-rabbit-igg-hl-hrp-ab205722.pdf

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Immunology Immunoglobulins Heavy Chain IgG
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Donkey Anti-Rabbit IgG H&L (HRP) (ab205722)

  • Datasheet
  • SDS
Reviews (2) Submit a question References (8)

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Western blot - Donkey Anti-Rabbit IgG H&L (HRP) (ab205722)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Donkey Anti-Rabbit IgG H&L (HRP) (ab205722)
  • Western blot - Donkey Anti-Rabbit IgG H&L (HRP) (ab205722)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Donkey Anti-Rabbit IgG H&L (HRP) (ab205722)
  • Western blot - Donkey Anti-Rabbit IgG H&L (HRP) (ab205722)
  • Western blot - Donkey Anti-Rabbit IgG H&L (HRP) (ab205722)
  • Western blot - Donkey Anti-Rabbit IgG H&L (HRP) (ab205722)
  • ELISA - Donkey Anti-Rabbit IgG H&L (HRP) (ab205722)

Key features and details

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

Conjugates logo Related conjugates and formulations

Alexa Fluor® 405 Alexa Fluor® 488 Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 568 Alexa Fluor® 594 Alexa Fluor® 647 Alexa Fluor® 680 Alexa Fluor® 750 Alexa Fluor® 790 Alkaline Phosphatase Biotin DyLight® 488 DyLight® 550 DyLight® 594 DyLight® 650 FITC Gold 10nm Gold 12nm Gold 18nm Gold 25nm Gold 6nm IRDye® 680RD Texas Red ® TRITC Unconjugated Unconjugated

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Overview

  • Product name

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

    Donkey
  • Target species

    Rabbit
  • Specificity

    The antibody used for conjugation reacts with rabbit immunoglobulins of all classes. Cross-reactions as determined by ELISA for the unconjugated antibody (ab182020): Human IgG, mouse IgG, rat IgG, goat IgG and chicken IgY, less than 2%.
  • Tested applications

    Suitable for: IHC-P, IP, WB, ELISAmore 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-Rabbit
    • IgG
    • Enzyme
    • HRP

Associated products

  • Alternative Versions

    • Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150073)
    • Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 555) (ab150074)
    • Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 647) (ab150075)
    • Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150076)
    • Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 568) (ab175470)
    • Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 405) (ab175651)
    • Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 750) (ab175731)
    • Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 680) (ab175772)
    • Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 790) (ab175780)
    • Donkey Anti-Rabbit IgG H&L (ab182020)
    • Donkey Anti-Rabbit IgG H&L (Biotin) (ab207999)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab205722 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
1/2000 - 1/20000.
IP
Use at an assay dependent concentration.
WB (2)
1/2000 - 1/50000.
ELISA
Use at an assay dependent concentration.
Notes
IHC-P
1/2000 - 1/20000.
IP
Use at an assay dependent concentration.
WB
1/2000 - 1/50000.
ELISA
Use at an assay dependent concentration.

Images

  • Western blot - Donkey Anti-Rabbit IgG H&L (HRP) (ab205722)
    Western blot - Donkey Anti-Rabbit IgG H&L (HRP) (ab205722)
    All lanes : Anti-beta Actin antibody (ab8227) 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-Rabbit IgG H&L (HRP) (ab205722) at 1/10000 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 ab8227 overnight at 4°C. Antibody binding was detected using ab205722, and visualised using ECL development solution ab133406.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Donkey Anti-Rabbit IgG H&L (HRP) (ab205722)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Donkey Anti-Rabbit IgG H&L (HRP) (ab205722)

    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 ab177840 at 1ug/ml. An HRP-conjugated secondary (Ab205722, 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

  • Western blot - Donkey Anti-Rabbit IgG H&L (HRP) (ab205722)
    Western blot - Donkey Anti-Rabbit IgG H&L (HRP) (ab205722)
    All lanes : Anti-beta Actin antibody (ab8227) at 1 µg/ml

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

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : ab205722 (Left Image) at 1/10,000 and a competitor secondary (Right Image) at 1/10,000. Notice the decreased signal 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: 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 ab8227 overnight at 4°C. Antibody binding was detected using ab205722 (Left Image) and a competitor secondary (Right Image), and visualised using ECL development solution ab133406.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Donkey Anti-Rabbit IgG H&L (HRP) (ab205722)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Donkey Anti-Rabbit IgG H&L (HRP) (ab205722)

    IHC image of beta 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 ab6046 at 5ug/ml. An HRP-conjugated secondary (Ab205722, 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

  • Western blot - Donkey Anti-Rabbit IgG H&L (HRP) (ab205722)
    Western blot - Donkey Anti-Rabbit IgG H&L (HRP) (ab205722)
    All lanes : No Primary Antibody

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

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : ab205722 (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 ab205722 (Left Image) and a competitor secondary (Right Image), and visualised using ECL development solution ab133406.

  • Western blot - Donkey Anti-Rabbit IgG H&L (HRP) (ab205722)
    Western blot - Donkey Anti-Rabbit IgG H&L (HRP) (ab205722)
    All lanes : Anti-STAT3 antibody [EPR787Y] (ab68153) at 1/2000 dilution

    Lane 1 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Heart (Mouse) Tissue Lysate
    Lane 3 : Heart (Rat) Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Donkey Anti-Rabbit IgG H&L (HRP) (ab205722) at 1/2000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Observed band size: 88 kDa why is the actual band size different from the predicted?


    Exposure time: 20 minutes


    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 3% milk before being incubated with ab68153 overnight at 4°C. Antibody binding was detected using ab205722, and visualised using ECL development solution ab133406.

  • Western blot - Donkey Anti-Rabbit IgG H&L (HRP) (ab205722)
    Western blot - Donkey Anti-Rabbit IgG H&L (HRP) (ab205722)
    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-Rabbit IgG H&L (HRP) (ab205722) 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 (ab205722), and visualised using ECL development solution ab133406.

  • ELISA - Donkey Anti-Rabbit IgG H&L (HRP) (ab205722)
    ELISA - Donkey Anti-Rabbit IgG H&L (HRP) (ab205722)

    Cross-reactivity of the polyclonal secondary antibody ab182020 was tested using a sandwich ELISA approach. The wells were coated with the indicated IgG standards at 1 µg/ml (50 µl/well) and incubated overnight at 4°C, followed by a 5% BSA blocking step for 2h at RT. ab182020 was then added starting at 1 µg/ml and gradually diluted 1/4 (50 µl/well), followed by incubation for 2h. For the detection Goat anti-Donkey IgG H&L (HRP) (ab6988) was used at 1/20,000 dilution (50 µl/well), followed by incubation for 1h at RT.

    For the batch tested, ab182020 showed a cross-reactivity below 2% towards human IgG, mouse IgG, rat IgG, goat IgG and chicken IgY.

    This data was developed using the unconjugated antibody (ab182020).

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 (8)

Publishing research using ab205722? Please let us know so that we can cite the reference in this datasheet.

ab205722 has been referenced in 8 publications.

  • Liskiewicz AD  et al. Proteomic and Structural Manifestations of Cardiomyopathy in Rat Models of Obesity and Weight Loss. Front Endocrinol (Lausanne) 12:568197 (2021). PubMed: 33716957
  • Richards C  et al. Characterisation of cardiac health in the reduced uterine perfusion pressure model and a 3D cardiac spheroid model, of preeclampsia. Biol Sex Differ 12:31 (2021). PubMed: 33879252
  • Taha A  et al. Omega-3 and Omega-6 polyunsaturated fatty acids stimulate vascular differentiation of mouse embryonic stem cells. J Cell Physiol 235:7094-7106 (2020). PubMed: 32020589
  • Xu J  et al. The inhibition of miR-126 in cell migration and invasion of cervical cancer through regulating ZEB1. Hereditas 156:11 (2019). PubMed: 31007650
  • Serra M  et al. Time-restricted feeding delays the emergence of the age-associated, neoplastic-prone tissue landscape. Aging (Albany NY) 11:3851-3863 (2019). PubMed: 31188781
  • Liskiewicz A  et al. Methamphetamine-associated cognitive decline is attenuated by neutralizing IL-1 signaling. Brain Behav Immun N/A:N/A (2019). PubMed: 30885840
  • 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). PubMed: 28395332
  • Liu T  et al. Telocytes as potential targets in a cyclophosphamide-induced animal model of premature ovarian failure. Mol Med Rep 14:2415-22 (2016). PubMed: 27485835

Customer reviews and Q&As

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

1-2 of 2 Abreviews or Q&A

CRY2 Western Blot

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Human Cell lysate - whole cell (Liver).
Loading amount: 5 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: CRY2 (ab38872) diluted 1:1000 in 1%BSA/0.2%OvaAlb/PBS 14 hours at 4°C.
Secondary antibody: ab205722 - Donkey Anti-Rabbit IgG H&L (HRP) diluted 1:5000 in TBST.
ECL prime: 5 seconds.
Predicted size: 67kDa.
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 Aug 12 2016

Test ab205722 - Donkey Anti-Rabbit IgG H&L (HRP)

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Excellent specificity.
Good sensitivity.
WB conditions:
- Cell lisates: 20 μg protein
- non-reducing
- Primary antibody: Rabbit anti-LaminB1, ab133741. Abcam. Dilution 1:5000
- Secondary antibody: 1:6000
- Developer: ECL Prime, Amersham, GE Helathcare Life Sciences, Product code RPN2232
- Exposure time: 1 second, 2nd 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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