Antibodies, Proteins, Kits and Reagents for Life Science

301 536 ₸ Product size

100 µl 301 536 ₸ 10 µl 120 614 ₸

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Anti-alpha Actinin/ACTN1 antibody [EP2527Y] (ab68194)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EP2527Y] to alpha Actinin/ACTN1
Suitable for: IP, ICC, WB
Knockout validated
Reacts with: Mouse, Rat, Human

Product name

Anti-alpha Actinin/ACTN1 antibody [EP2527Y]
See all alpha Actinin/ACTN1 primary antibodies
Description

Rabbit monoclonal [EP2527Y] to alpha Actinin/ACTN1
Host species

Rabbit

Tested Applications & Species

Application	Species
ICC	Human
IP	Mouse
WB	Mouse Rat Human

See all applications and species data

Immunogen

Synthetic peptide corresponding to Human alpha Actinin/ACTN1.
Positive control
- ICC: ioSkeletal Myocytes - Human iPSC-Derived Skeletal Myocytes (ab277612); IP: NIH/3T3 whole cell lysate; WB: HeLa, PC-12, NIH/3T3, and C6 whole cell lysates.
General notes
This product was previously labelled as alpha Actinin

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.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Form

Liquid
Storage instructions

Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage buffer

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 0.05% BSA, 40% Glycerol (glycerin, glycerine), 59% PBS
Concentration information loading...
Purity

Tissue culture supernatant
Clonality

Monoclonal
Clone number

EP2527Y
Isotype

IgG
Research areas

Immunoprecipitation - Anti-alpha Actinin/ACTN1 antibody [EP2527Y] (ab68194)

Purified ab68194 at 1:20 dilution (2µg) immunoprecipitating alpha Actinin/ACTN1 in NIH/3T3 whole cell lysate.
Lane 1 (input): NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate 10µg
Lane 2 (+): ab68194 + NIH/3T3 whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab68194 in NIH/3T3 whole cell lysate.
VeriBlot for IP Detection Reagent (HRP)(ab131366) (1:5000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 103 kDa
Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2527Y] (ab68194)

Anti-alpha Actinin/ACTN1 antibody [EP2527Y] (ab68194) at 1/10000 dilution (Purified) + HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 15 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

Predicted band size: 103 kDa
Observed band size: 103 kDa

Blocking buffer: 5% NFDM/TBST
Immunocytochemistry - Anti-alpha Actinin/ACTN1 antibody [EP2527Y] (ab68194)

Immunofluorescence staining of Actinin/ACTN1 using ab68194 in ioSkeletal Myocytes - Human iPSC-Derived Skeletal Myocytes (ab277612), which were differentiated for 3 (left panel), 5 (middle panel) and 10 days (right panel) post induction.

The cells were fixed with 100% MeOH (5 min) and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab68194 at 0.5 µg/mL and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin, at 1/1000 dilution. Cells were then incubated with ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150120, Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) preadsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown. Gamma is adjusted to 1.5 in all channels.
Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2527Y] (ab68194)

All lanes : Anti-alpha Actinin/ACTN1 antibody [EP2527Y] (ab68194) at 1/1000 dilution (Purified)

Lane 1 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate at 20 µg
Lane 2 : C6 (Rat glial tumor glial cell) whole cell lysate at 15 µg
Lane 3 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate at 15 µg

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 103 kDa
Observed band size: 103 kDa

Blocking buffer: 5% NFDM/TBST
Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2527Y] (ab68194)

All lanes : Anti-alpha Actinin/ACTN1 antibody [EP2527Y] (ab68194) at 1/2000 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : ACTN1 knockout HeLa cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 103 kDa
Observed band size: 103 kDa

Lanes 1- 2: Merged signal (red and green). Green - ab68194 observed at 103 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab68194 was shown to react with alpha Actinin/ACTN1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265610 (knockout cell lysate ab257337) was used. Wild-type HeLa and ACTN1 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. ab68194 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at a 1 in 2000 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.
Immunocytochemistry - Anti-alpha Actinin/ACTN1 antibody [EP2527Y] (ab68194)

Immunofluorescence staining of Actinin/ACTN1 using ab68194 in ioSkeletal Myocytes - Human iPSC-Derived Skeletal Myocytes (ab277612), which were differentiated for 3 (left panel), 5 (middle panel) and 10 days (right panel) post induction.

The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% PBS-Tween for 5 mins and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab68194 at 0.5 µg/mL and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin, at 1/1000 dilution. Cells were then incubated with ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150120, Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) preadsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown. Gamma is adjusted to 1.5 in all channels.
Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2527Y] (ab68194)

Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: Alpha Actinin/ACTN1 knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: NIH 3T3 whole cell lysate (20 µg)

Lanes 1 - 4: Merged signal (red and green). Green - ab68194 observed at 103 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab68194 was shown to specifically react with alpha Actinin/ACTN1 in wild-type HAP1 cells as signal was lost in alpha Actinin/ACTN1 knockout cells. Wild-type and alpha Actinin/ACTN1 knockout samples were subjected to SDS-PAGE. ab68194 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/2000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

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