Recombinant Anti-HuD + HuC antibody [EPR19098] (ab184267)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR19098] to HuD + HuC
- Suitable for: Flow Cyt (Intra), Indirect ELISA, WB, IHC-P, IHC-Fr, ICC/IF, IP
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-HuD + HuC antibody [EPR19098] -
Description
Rabbit monoclonal [EPR19098] to HuD + HuC -
Host species
Rabbit -
Specificity
This antibody shows very weak cross reactivity to HuB in WB test. Please contact our Scientific support team for more information.
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Tested applications
Suitable for: Flow Cyt (Intra), Indirect ELISA, WB, IHC-P, IHC-Fr, ICC/IF, IPmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Mouse His-tagged HuC and His-GST-tagged HuD recombinant proteins; Human fetal brain; Mouse and rat brain; Neuro-2a and SH-SY5Y cell lysates. IHC-P: Human cerebral cortex, colon and glioma tissues; Mouse cerebral cortex and stomach tissues; Rat cerebral cortex and colon tissues. IHC-Fr: Mouse cerebral cortex and stomach tissues; Rat cerebral cortex and colon tissues. ICC/IF: SH-SY5Y and Neuro-2a cells. Flow Cyt: SH-SY5Y and Neuro-2a cells. IP: Mouse brain lysate. I-ELISA: Mouse Elavl3 and 4.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
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 long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR19098 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab184267 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 |
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Flow Cyt (Intra) |
1/500.
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Indirect ELISA |
Use a concentration of 1 µg/ml.
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WB |
1/1000. Detects a band of approximately 39 kDa (predicted molecular weight: 40 kDa).
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IHC-P |
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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IHC-Fr |
1/500.
Antigen retrieval: Heated citrate solution (10mM citrate pH 6.0 + 0.05% Tween-20). |
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ICC/IF |
1/500.
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IP |
1/30.
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Notes |
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Flow Cyt (Intra)
1/500. |
Indirect ELISA
Use a concentration of 1 µg/ml. |
WB
1/1000. Detects a band of approximately 39 kDa (predicted molecular weight: 40 kDa). |
IHC-P
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
IHC-Fr
1/500. Antigen retrieval: Heated citrate solution (10mM citrate pH 6.0 + 0.05% Tween-20). |
ICC/IF
1/500. |
IP
1/30. |
Images
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All lanes : Anti-HuD + HuC antibody [EPR19098] (ab184267) at 1/1000 dilution
Lane 1 : His-tagged Mouse HUR recombinant protein (aa1-326) at 0.01 µg
Lane 2 : His-tagged Mouse HUR recombinant protein (aa1-326) at 0.05 µg
Lane 3 : His-tagged Mouse HUR recombinant protein (aa1-326) at 0.1 µg
Lane 4 : His-tagged mouse HuB recombinant protein (aa1-360) at 0.01 µg
Lane 5 : His-tagged mouse HuB recombinant protein (aa1-360) at 0.05 µg
Lane 6 : His-tagged mouse HuB recombinant protein (aa1-360) at 0.1 µg
Lane 7 : Blank
Lane 8 : His-tagged mouse HuC recombinant protein (aa1-367) at 0.01 µg
Lane 9 : His-GST-tagged mouse HuD recombinant protein (aa1-385) at 0.01 µg
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 40 kDa
Observed band size: 37,75 kDa why is the actual band size different from the predicted?
Exposure time: 10 secondsBlocking and diluting buffer and concentration: 5% NFDM/TBST.
ab184267 shows very weak cross reactivity with HuB.
Band around 80kda detected by anti-His tag antibody should be dimer.
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Indirect ELISA showing primary antibody ab184267 binding to Mouse Elavl1 (HuR); Mouse Elavl2 (HuB); Mouse Elavl3 (HuC); Mouse Elavl4 (HuD). Antigen concentration is 1000 ng/ml. Substrate solution is p-nitrophenyl phosphate(PNPP).
Binding of ab184267 was assessed in a serial dilution range 1000-0 ng/ml.
Binding was detected using the secondary antibody, Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) at 1/2500 dilution.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HuD + HuC antibody [EPR19098] (ab184267)
Immunohistochemical analysis of paraffin-embedded human cerebral cortex tissue labeling HuC + HuD with ab184267 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Nuclear and weakly cytoplasmic staining on neurons of human cerebral cortex is observed [PMID: 22007133].
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized SH-SY5Y (Human neuroblastoma cell line from bone marrow) cells labeling HuC + HuD with ab184267 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing cytoplasmic and nuclear staining on SH-SY5Y cell line.
The nuclear counterstain is DAPI (blue).
Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.
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Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen mouse cerebral cortex tissue labeling HuC + HuD with ab184267 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Cytoplasmic and nuclear staining on neurons of mouse cerebral cortex is observed.
The nuclear counterstain is DAPI (blue).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.
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All lanes : Anti-HuD + HuC antibody [EPR19098] (ab184267) at 1/2000 dilution
Lane 1 : His-tagged mouse HuC recombinant protein (aa1-367)
Lane 2 : His-GST-tagged mouse HuD recombinant protein (aa1-385)
Lane 3 : His-tagged mouse HuR recombinant protein (aa1-360)
Lane 4 : His-tagged mouse HuB recombinant protein (aa1-360)
Lysates/proteins at 0.01 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 40 kDa
Observed band size: 39,68 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 0.5 second.
Please note that the band around 55kDa in lane 2 is most likely the HuD protein with tags.
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Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed SH-SY5Y (Human neuroblastoma cell line from bone marrow) cells labeling HuC + HuDwith ab184267 at 1/500 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.
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HuC + HuD was immunoprecipitated from 0.35 mg of mouse brain lysate with ab184267 at 1/30 dilution.
Western blot was performed from the immunoprecipitate using ab184267 at 1/1000 dilution.
VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1: Mouse brain lysate, 10 µg (Input).
Lane 2: ab184267 IP in mouse brain lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab184267 in mouse brain lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 30 seconds.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Neuro-2a (Mouse neuroblastoma cell line) cells labeling HuC + HuD with ab184267 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing cytoplasmic and nuclear staining on Neuro-2a cell line.
The nuclear counterstain is DAPI (blue).
Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.
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All lanes : Anti-HuD + HuC antibody [EPR19098] (ab184267) at 1/1000 dilution
Lane 1 : Human fetal brain lysate
Lane 2 : Human fetal heart lysate
Lane 3 : Human fetal kidney lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/4000 dilution
Predicted band size: 40 kDa
Observed band size: 39 kDa why is the actual band size different from the predicted?
Exposure time: 8 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
The expression profile is consistent with what has been described in the literature (PMID: 9016658; PMID:8535975).
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All lanes : Anti-HuD + HuC antibody [EPR19098] (ab184267) at 1/1000 dilution
Lane 1 : Mouse brain lysate
Lane 2 : Mouse heart lysate
Lane 3 : Mouse kidney lysate
Lane 4 : Rat brain lysate
Lane 5 : Rat heart lysate
Lane 6 : Rat kidney lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 40 kDa
Observed band size: 39 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1-3: 1 minute; Lane 4-6: 5 seconds.
The expression profile is consistent with what has been described in the literature (PMID: 9016658; PMID: 8535975).
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All lanes : Anti-HuD + HuC antibody [EPR19098] (ab184267) at 1/5000 dilution
Lane 1 : Neuro-2a (Mouse neuroblastoma cell line) whole cell lysate
Lane 2 : SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 40 kDa
Observed band size: 39 kDa why is the actual band size different from the predicted?
Exposure time: 5 secondsBlocking/Dilution Buffer: 5% NFDM/TBST
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HuD + HuC antibody [EPR19098] (ab184267)
Immunohistochemical analysis of paraffin-embedded human colon tissue labeling HuC + HuD with ab184267 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Nuclear and weakly cytoplasmic staining on myenteric ganglia of human colon is observed [PMID: 16918730].
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HuD + HuC antibody [EPR19098] (ab184267)
Immunohistochemical analysis of paraffin-embedded human glioma tissue labeling HuC + HuD with ab184267 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Nuclear and weakly cytoplasmic staining on human glioma is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HuD + HuC antibody [EPR19098] (ab184267)
Immunohistochemical analysis of paraffin-embedded mouse cerebral cortex tissue labeling HuC + HuD with ab184267 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Nuclear and weakly cytoplasmic staining on neurons of mouse cerebral cortex is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HuD + HuC antibody [EPR19098] (ab184267)
Immunohistochemical analysis of paraffin-embedded mouse stomach tissue labeling HuC + HuD with ab184267 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Cytoplasmic and nuclear staining on myenteric ganglia of mouse stomach is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HuD + HuC antibody [EPR19098] (ab184267)
Immunohistochemical analysis of paraffin-embedded rat cerebral cortex tissue labeling HuC + HuD with ab184267 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Nuclear and weakly cytoplasmic staining on neurons of rat cerebral cortex is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HuD + HuC antibody [EPR19098] (ab184267)
Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling HuC + HuD with ab184267 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Cytoplasmic and nuclear staining on myenteric ganglia of rat colon is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen mouse stomach tissue labeling HuC + HuD with ab184267 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Cytoplasmic and nuclear staining on myenteric ganglia of mouse stomach is observed (white arrow).
The nuclear counterstain is DAPI (blue).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.
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Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen rat cerebral cortex tissue labeling HuC + HuD with ab184267 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Cytoplasmic and nuclear staining on neurons of rat cerebral cortex is observed.
The nuclear counterstain is DAPI (blue).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.
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Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen rat colon tissue labeling HuC + HuD with ab184267 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Cytoplasmic and nuclear staining on myenteric ganglia of rat colon is observed.
The nuclear counterstain is DAPI (blue).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.
-
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed Neuro-2a (Mouse neuroblastoma cell line) cells labeling HuC + HuDwith ab184267 at 1/500 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (9)
ab184267 has been referenced in 9 publications.
- Wang Y et al. Prucalopride might improve intestinal motility by promoting the regeneration of the enteric nervous system in diabetic rats. Int J Mol Med 50:N/A (2022). PubMed: 35543167
- Li YR et al. Involvement of nitrergic neurons in colonic motility in a rat model of ulcerative colitis. World J Gastroenterol 28:3854-3868 (2022). PubMed: 36157548
- Chen W et al. Patchouli alcohol improved diarrhea-predominant irritable bowel syndrome by regulating excitatory neurotransmission in the myenteric plexus of rats. Front Pharmacol 13:943119 (2022). PubMed: 36452228
- Yuan H et al. Premigratory neural crest stem cells generate enteric neurons populating the mouse colon and regulating peristalsis in tissue-engineered intestine. Stem Cells Transl Med 10:922-938 (2021). PubMed: 33481357
- Zeng Y et al. The Impact of Particulate Matter (PM2.5) on Human Retinal Development in hESC-Derived Retinal Organoids. Front Cell Dev Biol 9:607341 (2021). PubMed: 33644046
- Yi J et al. Histone demethylase UTX/KDM6A enhances tumor immune cell recruitment, promotes differentiation and suppresses medulloblastoma. Cancer Lett 499:188-200 (2021). PubMed: 33253789
- Kalusa M et al. Developmental Differences in Neocortex Neurogenesis and Maturation Between the Altricial Dwarf Rabbit and Precocial Guinea Pig. Front Neuroanat 15:678385 (2021). PubMed: 34135738
- Lai NY et al. Gut-Innervating Nociceptor Neurons Regulate Peyer's Patch Microfold Cells and SFB Levels to Mediate Salmonella Host Defense. Cell 180:33-49.e22 (2020). PubMed: 31813624
- Illing RB et al. Mitotic activity, modulation of DNA processing, and purinergic signalling in the adult rat auditory brainstem following sensory deafferentation. Eur J Neurosci 50:3985-4003 (2019). PubMed: 31325398