Donkey Anti-Goat IgG H&L (HRP) (ab205723)
Key features and details
- Donkey Anti-Goat IgG H&L (HRP)
- Conjugation: HRP
- Host species: Donkey
- Isotype: IgG
- Suitable for: IP, WB, IHC-P, ELISA
Overview
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Product name
Donkey Anti-Goat IgG H&L (HRP)
See all IgG secondary antibodies -
Host species
Donkey -
Target species
Goat -
Specificity
The antibody used for conjugation reacts with goat immunoglobulins of all classes. Cross-reactions as determined by ELISA for the unconjugated antibody (ab182021): Human IgG, mouse IgG, rat IgG, rabbit IgG and chicken IgY, less than 2%. -
Tested applications
Suitable for: IP, WB, IHC-P, ELISAmore details -
Immunogen
Full length protein corresponding to Goat IgG.
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Conjugation
HRP
Properties
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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% 10% Proclin 300 Solution
Constituents: PBS, 1% BSA, 30% Glycerol -
Concentration information loading...
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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
Associated products
Applications
Our Abpromise guarantee covers the use of ab205723 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 |
---|---|---|
IP | Use at an assay dependent concentration. | |
WB | 1/2000 - 1/20000. | |
IHC-P | 1/2000 - 1/20000. | |
ELISA | Use at an assay dependent concentration. |
Images
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All lanes : Anti-beta Actin antibody - Loading Control (ab8229) 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-Goat IgG H&L (HRP) (ab205723) 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: 30 secondsThis 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 ab8229 overnight at 4°C. Antibody binding was detected using ab205723, and visualised using ECL development solution ab133406.
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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-Goat IgG H&L (HRP) (ab205723) at 1/2000 dilution
Performed under reducing conditions.
Exposure time: 5 secondsThis 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 (ab205723), and visualised using ECL development solution ab133406.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Donkey Anti-Goat IgG H&L (HRP) (ab205723)
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 1ug/ml. Ab98800, goat anti-mouse IgG, was then added as a secondary bridging antibody, at 1/250 dilution for 1h.
An HRP-conjugated secondary (Ab205723, 1/2000 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
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.
References (9)
ab205723 has been referenced in 9 publications.
- Cai X et al. miR-124a enhances therapeutic effects of bone marrow stromal cells transplant on diabetic nephropathy-related epithelial-to-mesenchymal transition and fibrosis. J Cell Biochem 121:299-312 (2020). PubMed: 31190436
- Pan J et al. lncRNA ZFAS1 Is Involved in the Proliferation, Invasion and Metastasis of Prostate Cancer Cells Through Competitively Binding to miR-135a-5p. Cancer Manag Res 12:1135-1149 (2020). PubMed: 32104094
- Giraud E et al. Osteopontin in the host response to Leishmania amazonensis. BMC Microbiol 19:32 (2019). PubMed: 30736736
- Fu Q et al. Analysis of clinical characteristics of macrophage capping protein (CAPG) gene expressed in glioma based on TCGA data and clinical experiments. Oncol Lett 18:1344-1350 (2019). PubMed: 31423196
- Su T et al. LIM homeodomain transcription factor Isl1 affects urethral epithelium differentiation and apoptosis via Shh. Cell Death Dis 10:713 (2019). PubMed: 31558700
- Zhou R et al. Recombinant CC16 inhibits NLRP3/caspase-1-induced pyroptosis through p38 MAPK and ERK signaling pathways in the brain of a neonatal rat model with sepsis. J Neuroinflammation 16:239 (2019). PubMed: 31775794
- Mao X et al. Up-regulated Linc00472 suppresses development of lung cancer cell via inhibition of MiR-196b-5p. Biosci Biotechnol Biochem N/A:1-13 (2019). PubMed: 31791206
- Bhargava R et al. C-NHEJ without indels is robust and requires synergistic function of distinct XLF domains. Nat Commun 9:2484 (2018). PubMed: 29950655
- Zhou Z et al. siRNA targeting YAP gene inhibits gastric carcinoma growth and tumor metastasis in SCID mice. Oncol Lett 11:2806-2814 (2016). IHC . PubMed: 27073556