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Immunology Immunoglobulins Heavy Chain IgG
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Goat Anti-Rabbit IgG H&L (HRP) (ab205718)

  • Datasheet
  • SDS
Reviews (6) Submit a question References (567)

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Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • ELISA - Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • ELISA - Goat Anti-Rabbit IgG H&L (HRP) (ab205718)

Key features and details

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

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Overview

  • Product name

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

    Goat
  • 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 (ab182016): Mouse IgG, rat IgG, and chicken IgY, less than 2%. Human IgG, less than 7%.
  • Tested applications

    Suitable for: IHC-P, WB, ELISA, IPmore details
  • Immunogen

    . Rabbit IgG, whole molecule

  • 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 long term. Avoid freeze / thaw cycle. Store In the Dark.
  • Storage buffer

    pH: 7.40
    Preservative: 0.1% 10% 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

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 555) (ab150078)
    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) (ab150079)
    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080)
    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 568) (ab175471)
    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 405) (ab175652)
    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 680) (ab175773)
    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 790) (ab175781)
    • Goat Anti-Rabbit IgG H&L (ab182016)
    • Goat Anti-Rabbit IgG H&L (Biotin) (ab207995)
  • Related Products

    • Normal Goat Serum (ab7481)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab205718 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 - 1/50000.
WB (5)
1/2000 - 1/50000.
ELISA
1/5000 - 1/20000.
IP
Use at an assay dependent concentration.
Notes
IHC-P
1/2000 - 1/50000.
WB
1/2000 - 1/50000.
ELISA
1/5000 - 1/20000.
IP
Use at an assay dependent concentration.

Images

  • Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    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 : Goat Anti-Rabbit IgG H&L (HRP) (ab205718) at 1/50000 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: 30 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 ab205718, and visualised using ECL development solution ab133406.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Rabbit IgG H&L (HRP) (ab205718)

    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 1/1000 dilution. An HRP-conjugated secondary (Ab205718, 1/20000 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 - Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    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 : ab205718 (Left Image) at 1/20,000 and a competitor secondary (Right Image) at 1/50,000. 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: 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 ab8227 overnight at 4°C. Antibody binding was detected using ab205718 (Left Image) and a competitor secondary (Right Image), and visualised using ECL development solution ab133406.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Rabbit IgG H&L (HRP) (ab205718)

    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 1/100 dilution. An HRP-conjugated secondary (Ab205718, 1/20000 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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Rabbit IgG H&L (HRP) (ab205718)

    IHC image of Ki67 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 ab15580 at 1/1000 dilution. An HRP-conjugated secondary (Ab205718, 1/20000 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

  • ELISA - Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    ELISA - Goat Anti-Rabbit IgG H&L (HRP) (ab205718)

    Cross-reactivity of the polyclonal secondary antibody ab182016 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. ab182016 was then added starting at 1 µg/ml and gradually diluted 1/4 (50 µl/well), followed by incubation for 2h. For the detection Donkey anti-Goat IgG H&L (HRP) (ab6885) was used at 1/10,000 dilution (50 µl/well), followed by incubation for 1h at RT.

    For the batch tested, ab182016 showed a cross-reactivity of 5-7% towards Human IgG and below 2% towards Mouse IgG, Rat IgG and Chicken IgY.

    This data was developed using the unconjugated antibody (ab182016).

  • ELISA - Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    ELISA - Goat Anti-Rabbit IgG H&L (HRP) (ab205718)

    Cross-reactivity of Goat anti-Rabbit IgG H&L (ab182016) and Goat anti-Rabbit IgG H&L obtained from two different vendors was tested using a sandwich ELISA approach. The wells were coated with the indicated IgG standards (Rabbit, Human, Mouse and Rat) at 1 µg/ml (50 µl/well) and incubated overnight at 4°C, followed by a 5% BSA blocking step for 2h at RT. Secondary antibodies were then added starting at 1 µg/ml and gradually diluted 1/4 (50 µl/well), followed by incubation for 2h. For the detection Donkey anti-Goat IgG H&L (HRP) (ab6885) was used at 1/10,000 dilution (50 µl/well), followed by incubation for 1h at RT. This data is from a representative dilution.

    This data was developed using the unconjugated antibody (ab182016).

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 (567)

Publishing research using ab205718? Please let us know so that we can cite the reference in this datasheet.

ab205718 has been referenced in 567 publications.

  • Zhang J  et al. MicroRNA-132 protects H9c2 cells against oxygen and glucose deprivation-evoked injury by targeting FOXO3A. J Cell Physiol 235:176-184 (2020). PubMed: 31210352
  • Xi X  et al. MicroRNA-204-3p represses colon cancer cells proliferation, migration, and invasion by targeting HMGA2. J Cell Physiol 235:1330-1338 (2020). PubMed: 31286521
  • Zhao DY  et al. Ligustilide protects PC12 cells from oxygen-glucose deprivation/reoxygenation-induced apoptosis via the LKB1-AMPK-mTOR signaling pathway. Neural Regen Res 15:473-481 (2020). PubMed: 31571659
  • Dai L  et al. Knockdown of long non-coding RNA LINC00176 suppresses ovarian cancer progression by BCL3-mediated down-regulation of ceruloplasmin. J Cell Mol Med 24:202-213 (2020). PubMed: 31668012
  • Shang D  et al. Pancreatic cancer cell-derived exosomal microRNA-27a promotes angiogenesis of human microvascular endothelial cells in pancreatic cancer via BTG2. J Cell Mol Med 24:588-604 (2020). PubMed: 31724333
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review

Filter by Application

Filter by Ratings

1-6 of 6 Abreviews

EXCELLENT SECONDARY ANTIBODY for WB

Excellent
Abreviews
Abreviews
Application
Western blot
EXCELLENT SECONDARY ANTIBODY for WB

Abcam user community

Verified customer

Submitted Jul 10 2020

Anti-rabbit secondary successful IHC-P

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry paraffin embedded sections
The first image was incubated with ab40763 and ab205718 (at 1/200)
The second image is a negative control, with no primary antibody and ab205718 (at /200).

Abcam user community

Verified customer

Submitted Jun 08 2020

ab205718 tested by WB application

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Assay performed on whole cell lysate (mouse embryonic stem cells). Samples were mixed with 2X Laemli sample buffer and loaded on a 8% acrylamide gel together with Spectra™ Multicolor High Range Protein Ladder. Run performed in vertical tank at 100V for 1 hour followed by transfer in a wet system for 2 hours at 60V. The membrane was preblocked with 5% milk and then incubated with primary antibody (alpha-tubulin) overnight diluted in 5%milk. After washes the membrane was incubated with ab205718 1:2000 for 2 hours at RT. Bands were detected with ECL.
First lane:ladder, second lane:wild type mouse ES cells (D3), third lane: E-cadherin knock-out mouse embryonic cell line.
ab205718 worked for the chosen protein by WB. However, additional unspecific band detected.

Miss. Fabiana Luise

Verified customer

Submitted Jul 24 2019

Goat Anti-Rabbit IgG with AMPK alpha antibody (ab32047)

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
- 20ug of bovine liver lysates were loaded on to 4-15% gradient gel.
- Blocking in 5% skim milk for an hour at room temperature.
- 1:2000 AMPK alpha antibody (ab32047) were incubated over night @ 4C
- 1:20000 secondary antibody (ab205718) used for an hour at room temperature.

Julie Kim

Verified customer

Submitted Apr 20 2017

NR1D1 Western blot

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
10ug whole cell lysate (human ES cells).
Blocking buffer - 5%BSA/1%OvAlb/PBS for 3 hours.
Primary antibody - NR1D1(ab174309) 1:1000, 15 hours at 4C, expected size 67kDa.
Secondary diluted in TBST (1:5000) for 3 hours at room temperature.
ECL prime: 1 second exposure.

Abcam user community

Verified customer

Submitted Aug 12 2016

Test ab205718 - Goat Anti-Rabbit IgG H&L (HRP)

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Excellent specificity.
Excellent 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

Abcam user community

Verified customer

Submitted Feb 01 2016

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