Anti-HAND2 antibody [EPR19451] (ab200040)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR19451] to HAND2
- Suitable for: Flow Cyt, IHC-P, WB, ICC/IF, IP
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Anti-HAND2 antibody [EPR19451]
See all HAND2 primary antibodies -
Description
Rabbit monoclonal [EPR19451] to HAND2 -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species Flow Cyt MouseICC/IF MouseHumanIHC-P MouseRatIP HumanWB MouseHuman -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HEK-293T transfected with mouse HAND2 expression vector containing a myc tag, whole cell lysate; Neuro-2a and SH-SY5Y whole cell lysates. IHC-P: Mouse E14.5 embryo and E14.5 embryonic adrenal gland tissues; Rat E14.5 embryo and E14.5 embryonic gut tissues. ICC/IF: Neuro-2a and SH-SY5Y cells. Flow Cyt: Neuro-2a cells. IP: SH-SY5Y whole cell lysate.
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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 (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR19451 -
Isotype
IgG -
Research areas
Images
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Immunohistochemical analysis of paraffin-embedded mouse E14.5 embryo tissue labeling HAND2 with ab200040 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining in multiple tissues of E14.5 mouse embryo (PMID: 25248475). Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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All lanes : Anti-HAND2 antibody [EPR19451] (ab200040) at 1/1000 dilution
Lane 1 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with an empty vector (vector control) containing a myc tag, whole cell lysate
Lane 2 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with mouse HAND2 expression vector containing a myc tag, whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 24 kDa
Observed band size: 26 kDa why is the actual band size different from the predicted?
Exposure time: 1 secondBlocking/Dilution buffer: 5% NFDM/TBST.
The plasmid was kindly provided by Dr. Zhongzhou Yang from Nanjing University.
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All lanes : Anti-HAND2 antibody [EPR19451] (ab200040) at 1/1000 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 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 24 kDa
Observed band size: 25 kDa why is the actual band size different from the predicted?Exposure times: Lane 1: 3 minutes; Lane 2: 10 seconds.
Blocking/Dilution buffer: 5% NFDM/TBST.
The molecular mass observed is consistent with what has been described in the literature (PMID: 26166202); the upper band could be ubiquitinated Hand2 (PMID 21170031).
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Immunohistochemical analysis of paraffin-embedded mouse E14.5 embryonic adrenal gland tissue labeling HAND2 with ab200040 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining in mouse embryonic (E14.5) ganglion and adrenal gland (PMID: 25248475). Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
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 paraffin-embedded rat E14.5 embryonic tissue labeling HAND2 with ab200040 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining in multiple rat embryonic (E14.5) tissues (PMID: 25248475). Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
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 paraffin-embedded rat E14.5 embryonic gut tissue labeling HAND2 with ab200040 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining in rat E14.5 embryonic gut (PMID: 25248475). Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
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 Neuro-2a (mouse neuroblastoma cell line) cells labeling HAND2 with ab200040 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Nuclear staining in Neuro-2a cell line.
The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
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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 HAND2 with ab200040 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Nuclear staining in SH-SY5Y cell line.
The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
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Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized Neuro-2a (mouse neuroblastoma cell line) cell line labeling HAND2 with ab200040 at 1/400 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.
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HAND2 was immunoprecipitated from 0.35 mg of SH-SY5Y (human neuroblastoma cell line from bone marrow) whole cell lysate with ab200040 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab200040 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.
Lane 1: SH-SY5Y whole cell lysate 10 µg (Input).
Lane 2: ab200040 IP in SH-SY5Y whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab200040 in SH-SY5Y whole cell lysate.
Exposure time: Less than 1 second.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
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