Name: Anti-HuD + HuC antibody [EPR19098]
Brand: RabMAb
SKU: ab184267
Rating: 5 (2 reviews)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - 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), WB, IHC-P, IHC-Fr, ICC/IF, IP
Reacts with: Mouse, Rat, Human

Alexa Fluor® 488 Alexa Fluor® 647 Carrier Free

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.
Tested applications

Suitable for: Flow Cyt (Intra), 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.
Positive control
- WB: His-tagged mouse HuC recombinant protein; His-GST-tagged mouse HuD recombinant protein; Human fetal brain lysate; Mouse and rat brain lysates; 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.
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
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

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...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR19098
Isotype

IgG
Research areas

Alternative Versions
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Related Products
- VeriBlot for IP Detection Reagent (HRP) (ab131366)
- Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)

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
Flow Cyt (Intra)		1/500.
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.

Notes
Flow Cyt (Intra) 1/500.
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.

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.
Immunocytochemistry/ Immunofluorescence - Anti-HuD + HuC antibody [EPR19098] (ab184267)

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.
Immunohistochemistry (Frozen sections) - Anti-HuD + HuC antibody [EPR19098] (ab184267)

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.
Western blot - Anti-HuD + HuC antibody [EPR19098] (ab184267)

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.
Flow Cytometry (Intracellular) - Anti-HuD + HuC antibody [EPR19098] (ab184267)

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.
Immunoprecipitation - Anti-HuD + HuC antibody [EPR19098] (ab184267)

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.
Immunocytochemistry/ Immunofluorescence - Anti-HuD + HuC antibody [EPR19098] (ab184267)

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.
Western blot - Anti-HuD + HuC antibody [EPR19098] (ab184267)

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 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.

The expression profile is consistent with what has been described in the literature (PMID: 9016658; PMID:8535975).
Western blot - Anti-HuD + HuC antibody [EPR19098] (ab184267)

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).
Western blot - Anti-HuD + HuC antibody [EPR19098] (ab184267)

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 seconds

Blocking/Dilution Buffer: 5% NFDM/TBST
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.
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.
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.
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.
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.
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.
Immunohistochemistry (Frozen sections) - Anti-HuD + HuC antibody [EPR19098] (ab184267)

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.
Immunohistochemistry (Frozen sections) - Anti-HuD + HuC antibody [EPR19098] (ab184267)

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.
Immunohistochemistry (Frozen sections) - Anti-HuD + HuC antibody [EPR19098] (ab184267)

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.
Flow Cytometry (Intracellular) - Anti-HuD + HuC antibody [EPR19098] (ab184267)

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.
Anti-HuD + HuC antibody [EPR19098] (ab184267)

Flow cytometry protocols
Immunoprecipitation protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

SDS download

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Datasheet download

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Publishing research using ab184267? Please let us know so that we can cite the reference in this datasheet.

ab184267 has been referenced in 4 publications.

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
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

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1-2 of 2 Abreviews or Q&A

Application

Immunocytochemistry

Sample

Human Cell (Human neurons)

Permeabilization

Yes - Triton X-100

Specification

Human neurons

Blocking step

Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

Fixative

Paraformaldehyde

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 Nov 24 2021

Application

IHC - Wholemount

Sample

Rat Tissue (Jejunum myenteric plexus)

Specification

Jejunum myenteric plexus

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 Nov 25 2019

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Specificity

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Related Products

Applications

The Abpromise guarantee

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (4)

Customer reviews and Q&As

Immunocytochemistry abreview for Anti-HuD + HuC antibody [EPR19098]

IHC - Wholemount abreview for Anti-HuD + HuC antibody [EPR19098]