Antibodies, Proteins, Kits and Reagents for Life Science

361 843 ₸ Product size

100 µl 361 843 ₸ 10 µl 144 067 ₸

Order now and get it on Tuesday March 02, 2021

Anti-HDAC6 antibody [EPR1698(2)] (ab133493)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR1698(2)] to HDAC6
Suitable for: WB, IHC-P, Flow Cyt
Knockout validated
Reacts with: Human, African green monkey

Product name

Anti-HDAC6 antibody [EPR1698(2)]
See all HDAC6 primary antibodies
Description

Rabbit monoclonal [EPR1698(2)] to HDAC6
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Human
IHC-P	Human
WB	Human

See all applications and species data

Immunogen

Synthetic peptide within Human HDAC6 aa 1-100 (N terminal). The exact sequence is proprietary.
Database link: Q9UBN7
Positive control
- WB: HAP1, HeLa, Jurkat, and COS-1 cell lysates. Flow Cyt: K562. IHC-P: Human kidney tissue.
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. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant
Concentration information loading...
Purity

Tissue culture supernatant
Clonality

Monoclonal
Clone number

EPR1698(2)
Isotype

IgG
Research areas

Western blot - Anti-HDAC6 antibody [EPR1698(2)] (ab133493)

All lanes : Anti-HDAC6 antibody [EPR1698(2)] (ab133493) at 1/10000 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : HDAC6 knockout HeLa cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 131 kDa
Observed band size: 160 kDa
why is the actual band size different from the predicted?

Lanes 1- 2: Merged signal (red and green). Green - ab133493 observed at 160 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab133493 was shown to react with HDAC6 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab264804 (knockout cell lysate ab257145) was used. Wild-type HeLa and HDAC6 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab133493 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Flow Cytometry - Anti-HDAC6 antibody [EPR1698(2)] (ab133493)

ab133493 staining HDAC6 in K562 (human chronic myelogenous leukemia) by flow cytometry. Cells were fixed with 4% paraformaldehyde, permabilised with 90% methanol and the sample was incubated with the primary antibody at a dilution of 1/200. A goat anti rabbit IgG (Alexa Fluor^® 488) at a dilution of 1/2000 was used as the secondary antibody.

Isoytype control: Rabbit monoclonal IgG (Black)

Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC6 antibody [EPR1698(2)] (ab133493)

Immunohistochemical analysis of paraffin embedded Human kidney tissue labelling HDAC6 with ab133493 antibody at a dilution of 1/50.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Western blot - Anti-HDAC6 antibody [EPR1698(2)] (ab133493)

Lanes 1-2 : Anti-HDAC6 antibody [EPR1698(2)] (ab133493) at 1/10000 dilution
Lanes 3-4 : Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1/1000 dilution

Lanes 1 & 3 & 5 : Wild-type HAP1 cell lysate at 20 µg
Lanes 2 & 4 : HDAC6 knockout HAP1 cell lysate at 20 µg
Lane 6 : HDAC6 knockout HAP1 cell lysate

Predicted band size: 131 kDa

Lanes 1 and 2: Green signal from target – ab133493 observed at 160 kDa
Lanes 3 and 4: Red signal from loading control – ab8226 observed at 42 kDa
Lanes 5 and 6: Merged (red and green) signal

ab133493 was shown to specifically react with HDAC6 when HDAC6 knockout samples were used. Wild-type and HDAC6 knockout samples were subjected to SDS-PAGE. ab133493 and ab8226 (loading control to beta actin) were diluted 1/10 000 and 1/1000 respectively and incubated overnight at 4ºC. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.
Western blot - Anti-HDAC6 antibody [EPR1698(2)] (ab133493)

All lanes : Anti-HDAC6 antibody [EPR1698(2)] (ab133493) at 1/10000 dilution

Lane 1 : HeLa cell lysate
Lane 2 : Jurkat cell lysate
Lane 3 : K562 cell lysate
Lane 4 : COS-1 cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 131 kDa
Observed band size: 160 kDa why is the actual band size different from the predicted?
Anti-HDAC6 antibody [EPR1698(2)] (ab133493)

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