Antibodies, Proteins, Kits and Reagents for Life Science

304 886 ₸ Product size

100 µl 304 886 ₸

Order now and get it on Tuesday March 02, 2021

Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EP2528Y] to alpha Actinin/ACTN1
Suitable for: WB
Knockout validated
Reacts with: Mouse, Rat, Human

Product name

Anti-alpha Actinin/ACTN1 antibody [EP2528Y]
See all alpha Actinin/ACTN1 primary antibodies
Description

Rabbit monoclonal [EP2528Y] to alpha Actinin/ACTN1
Host species

Rabbit
Tested Applications & Species

Application Species
WB
Mouse

Rat

Human

See all applications and species data
Immunogen

Synthetic peptide within Human alpha Actinin/ACTN1 aa 750-850 (C terminal). The exact sequence is proprietary.
Positive control
- WB: HAP1, A431, NIH 3T3, C2C12, C6 and A673 cell lysates and human skeletal muscle and human heart tissue 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.

Application	Species
WB	Mouse Rat Human

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
Storage buffer

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

Protein A purified
Clonality

Monoclonal
Clone number

EP2528Y
Isotype

IgG
Research areas

Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265)

All lanes : Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265) at 1/1000 dilution

Lane 1 : Wild-type HAP1 cell lysate
Lane 2 : ACTN1 knockout HAP1 cell lysate
Lane 3 : A431 cell lysate
Lane 4 : Daudi cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 103 kDa
Observed band size: 115 kDa
why is the actual band size different from the predicted?

Lanes 1 - 4: Merged signal (red and green). Green - ab81265 observed at 115 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.

ab81265 was shown to react with ACTN1 in wild-type HAP1 cells in western blot with loss of signal observed in ACTN1 knockout sample. Wild-type and ACTN1 knockout HAP1 cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab81265 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4°C at a 1 in 1000 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265)

Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: Alpha Actinin 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 - ab81265 observed at 103 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab81265 was shown to recognize alpha Actinin in wild-type HAP1 cells as signal was lost at the expected MW in alpha Actinin knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and alpha Actinin knockout samples were subjected to SDS-PAGE. ab81265 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/500 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265)

All lanes : Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265) at 1/1000 dilution (purified)

Lane 1 : NIH/3T3 whole cell lysate
Lane 2 : C2C12 whole cell lysate
Lane 3 : C6 whole cell lysate

Lysates/proteins at 20 µg per lane.

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 and dilution buffer: 5% NFDM/TBST
Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265)

Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265) at 1/1000 dilution (purified) + A673 whole cell lysate at 20 µg

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

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

Blocking and dilution buffer: 5% NFDM/TBST
Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265)

All lanes : Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265) at 1/1000 dilution (unpurified)

Lane 1 : NIH 3T3 cell lysate
Lane 2 : human skeletal muscle tissue lysate
Lane 3 : human heart tissue lysate

Lysates/proteins at 10 µg per lane.

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

Predicted band size: 103 kDa
Observed band size: 103 kDa
Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265)

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