Antibodies, Proteins, Kits and Reagents for Life Science

281 433 ₸ Product size

100 µl 281 433 ₸

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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

Product name

Anti-HAND2 antibody [EPR19451]
See all HAND2 primary antibodies
Description

Rabbit monoclonal [EPR19451] to HAND2
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Mouse
ICC/IF	Mouse Human
IHC-P	Mouse Rat
IP	Human
WB	Mouse Human

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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.
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 (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR19451
Isotype

IgG
Research areas

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HAND2 antibody [EPR19451] (ab200040)

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.
Western blot - Anti-HAND2 antibody [EPR19451] (ab200040)

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 second

Blocking/Dilution buffer: 5% NFDM/TBST.

The plasmid was kindly provided by Dr. Zhongzhou Yang from Nanjing University.
Western blot - Anti-HAND2 antibody [EPR19451] (ab200040)

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).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HAND2 antibody [EPR19451] (ab200040)

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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HAND2 antibody [EPR19451] (ab200040)

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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HAND2 antibody [EPR19451] (ab200040)

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.
Immunocytochemistry/ Immunofluorescence - Anti-HAND2 antibody [EPR19451] (ab200040)

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.
Immunocytochemistry/ Immunofluorescence - Anti-HAND2 antibody [EPR19451] (ab200040)

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.
Flow Cytometry - Anti-HAND2 antibody [EPR19451] (ab200040)

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.
Immunoprecipitation - Anti-HAND2 antibody [EPR19451] (ab200040)

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.
Anti-HAND2 antibody [EPR19451] (ab200040)

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