Antibodies, Proteins, Kits and Reagents for Life Science

355 142 ₸ Product size

100 µl 355 142 ₸ 10 µl 144 067 ₸

Order now and get it on Tuesday March 09, 2021

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [Y415] to HDAC3
Suitable for: Flow Cyt, WB, IHC-P, ICC/IF, IP
Reacts with: Mouse, Rat, Human

Product name

Anti-HDAC3 antibody [Y415]
See all HDAC3 primary antibodies
Description

Rabbit monoclonal [Y415] to HDAC3
Host species

Rabbit
Specificity

This antibody may detect splice isoform 2 (RPD3-2A).

Tested Applications & Species

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

See all applications and species data

Immunogen

Synthetic peptide within Human HDAC3 (C terminal). The exact sequence is proprietary.
(Peptide available as ab221015)
Positive control
- K562 cell lysate; HeLa cells; human ovary carcinoma
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.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

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.
Storage buffer

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.12% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

Y415
Isotype

IgG
Research areas

Western blot - Anti-HDAC3 antibody [Y415] (ab32369)

All lanes : Anti-HDAC3 antibody [Y415] (ab32369) at 1/5000 dilution

Lane 1 : K562 (human chronic myelogenous leukemia) whole cell lysate
Lane 2 : HeLa (human cervix adenocarcinoma) whole cell lysate
Lane 3 : NIH/3T3 (mouse embryo) whole cell lysate
Lane 4 : C6 (rat glioma) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

Predicted band size: 49 kDa
Observed band size: 49 kDa

Blocking/Diluting buffer 5% NFDM/TBST
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC3 antibody [Y415] (ab32369)

Immunohistochemical analysis of paraffin-embedded human endometrial carcinoma sections labelling HDAC3 with purified ab32369 at dilution of 1:50. The secondary antibody used was ab97051; a goat anti-rabbit IgG H&L (HRP) at dilution of 1/500. The sample was counterstained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.
Immunocytochemistry/ Immunofluorescence - Anti-HDAC3 antibody [Y415] (ab32369)

Immunocytochemistry/Immunofluorescence analysis of K562 cells labelling HDAC3 with purified ab32369 at 1/500. Cells were fixed with 4% Paraformaldehyde and permeabilized using 0.1% Triton X-100. ab150077, Alexa Fluor^®488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Cells were co-stained with ab7291, a mouse anti-tubulin antibody (1/1000) using ab150120, an Alexa Fluor^®594-conjugated goat anti-mouse IgG (1/1000) as the secondary. Nuclei were counterstained with DAPI (blue).

For negative control 1, rabbit primary antibody and anti-mouse secondary antibody (ab150120) were used and for negative control 2, mouse primary antibody (ab7291) and anti-rabbit secondary antibody (ab150077) were used.
Flow Cytometry - Anti-HDAC3 antibody [Y415] (ab32369)

Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling HDAC3 with purified ab32369 at 1/30 dilution (10ug/mL) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor^®488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.
Immunocytochemistry/ Immunofluorescence - Anti-HDAC3 antibody [Y415] (ab32369)

Immunofluorescent staining of HeLa cells
using ab32369 at 1/250 dilution
Immunoprecipitation - Anti-HDAC3 antibody [Y415] (ab32369)

Purified ab32369 at 1/20 immunoprecipitating HDAC3 in K562 whole cell lysate observed at 49 KDa (lanes 1 and 2).

Lane 1 (input): K562 whole cell lysate 10ug

Lane 2 (+): ab32369 + K562 whole cell lysate.

Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32369 in K562 whole cell lysate

The secondary antibody used was VeriBlot for IP Detection Reagent (HRP) (ab131366) for detection at dilution of 1/1000.

Blocking/Diluting buffer 5% NFDM/TBST.
Western blot - Anti-HDAC3 antibody [Y415] (ab32369)

Anti-HDAC3 antibody [Y415] (ab32369) at 1/10000 dilution + K562 cell lysate

Predicted band size: 49 kDa
Observed band size: 49 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC3 antibody [Y415] (ab32369)

Paraffin-embedded human ovary carcinoma
ab32369 at 1/250 dilution

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Anti-HDAC3 antibody [Y415] (ab32369)

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