Name: Anti-alpha Actinin 4 antibody [EPR2533(2)]
Brand: RabMAb
SKU: ab108198
Rating: 5 (9 reviews)

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Western blot - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR2533(2)] to alpha Actinin 4
Suitable for: WB, IP, IHC-P, ICC/IF, Flow Cyt (Intra)
Knockout validated
Reacts with: Mouse, Rat, Human

Alexa Fluor® 488 Alexa Fluor® 647 Carrier Free HRP PE

Product name

Anti-alpha Actinin 4 antibody [EPR2533(2)]
See all alpha Actinin 4 primary antibodies
Description

Rabbit monoclonal [EPR2533(2)] to alpha Actinin 4
Host species

Rabbit
Tested applications

Suitable for: WB, IP, IHC-P, ICC/IF, Flow Cyt (Intra)more details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Human skeletal muscle, A431, Mouse brain, Rat brain, Rat Heart, HAP1 and MFC7 Lysates Flow Cyt: HeLa IHC-P: Human breast carcinoma, Mouse and Rat liver tissues ICC/IF: HAP1 cells (HAP1-ACTN4 knockout cells used as negative cell line), MCF7 IP: HeLa 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.
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. 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

EPR2533(2)
Isotype

IgG
Research areas

Alternative Versions
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Recombinant Protein
- Recombinant Human alpha Actinin 4 protein (ab126006)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab108198 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application	Abreviews	Notes
WB	(4)	1/1000 - 1/10000. Detects a band of approximately 100 kDa (predicted molecular weight: 105 kDa).
IP		1/30.
IHC-P		1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF	(3)	1/100 - 1/250.
Flow Cyt (Intra)		1/50. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Notes
WB 1/1000 - 1/10000. Detects a band of approximately 100 kDa (predicted molecular weight: 105 kDa).
IP 1/30.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF 1/100 - 1/250.
Flow Cyt (Intra) 1/50. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Function

F-actin cross-linking protein which is thought to anchor actin to a variety of intracellular structures. This is a bundling protein. Probably involved in vesicular trafficking via its association with the CART complex. The CART complex is necessary for efficient transferrin receptor recycling but not for EGFR degradation.
Tissue specificity

Widely expressed.
Involvement in disease

Defects in ACTN4 are the cause of focal segmental glomerulosclerosis type 1 (FSGS1) [MIM:603278]. A renal pathology defined by the presence of segmental sclerosis in glomeruli and resulting in proteinuria, reduced glomerular filtration rate and edema. Renal insufficiency often progresses to end-stage renal disease, a highly morbid state requiring either dialysis therapy or kidney transplantation.
Sequence similarities

Belongs to the alpha-actinin family.
Contains 1 actin-binding domain.
Contains 2 CH (calponin-homology) domains.
Contains 2 EF-hand domains.
Contains 4 spectrin repeats.
Cellular localization

Nucleus. Cytoplasm. Localized in cytoplasmic mRNP granules containing untranslated mRNAs. Colocalizes with actin stress fibers. Nuclear translocation can be induced by the PI3 kinase inhibitor wortmannin or by cytochalasin D. Exclusively localized in the nucleus in a limited number of cell lines.
Target information above from: UniProt accession O43707 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 81 Human
- Entrez Gene: 60595 Mouse
- Entrez Gene: 63836 Rat
- Omim: 604638 Human
- SwissProt: O43707 Human
- SwissProt: P57780 Mouse
- SwissProt: Q9QXQ0 Rat
- Unigene: 270291 Human
- Unigene: 81144 Mouse
- Unigene: 15777 Rat
see all
Alternative names
- actinin 4 antibody
- Actinin alpha 4 antibody
- actinin4 antibody
- ACTN 4 antibody
- ACTN4 antibody
- ACTN4_HUMAN antibody
- Alpha-actinin-4 antibody
- DKFZp686K23158 antibody
- F actin cross linking protein antibody
- F-actin cross-linking protein antibody
- Focal segmental glomerulosclerosis 1 antibody
- FSGS 1 antibody
- FSGS antibody
- FSGS1 antibody
- Non muscle alpha actinin 4 antibody
- Non-muscle alpha-actinin 4 antibody
see all

Western blot - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198)

Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198) 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 1/20000 dilution

Predicted band size: 105 kDa
Observed band size: 105 kDa
Western blot - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198)

All lanes : Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198) at 1/1000 dilution (Purified)

Lane 1 : Human skeletal muscle lysate
Lane 2 : A431 (Human epidermoid carcinoma epithelial cell) whole cell lysate
Lane 3 : Mouse brain lysate
Lane 4 : Rat brain lysate
Lane 5 : Rat heart lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 105 kDa
Observed band size: 105 kDa
Flow Cytometry (Intracellular) - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198)

Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling alpha Actinin 4 with Purified ab108198 at 1:50 dilution (10 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1:2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).
Immunocytochemistry/ Immunofluorescence - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198)

Immunocytochemistry analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling alpha Actinin 4 with Purified ab108198 at 1:250 dilution (2.0 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat liver tissue sections labeling alpha Actinin 4 with Purified ab108198 at 1:150 dilution (3.29 µg/mL). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse liver tissue sections labeling alpha Actinin 4 with Purified ab108198 at 1:150 dilution (3.29 µg/mL). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue sections labeling alpha Actinin 4 with Purified ab108198 at 1:150 dilution (3.29 µg/mL). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.
Immunocytochemistry/ Immunofluorescence - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198)

ab108198 staining ACTN4 in wild-type HAP1 cells (top panel) and ACTN4 knockout HAP1 cells (bottom panel). The cells were fixed with PFA (10min), permeabilized with 0.1% Triton X-100 for 5 minutes 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 with ab108198 at 1/100 dilution and ab7291 (Tubulin) at 1/1000 dilution overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (ab150120) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Western blot - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198)

Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: ACTN4 (alpha Actinin 4) knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: MCF7 whole cell lysate (20 µg)

Lanes 1 - 4: Merged signal (red and green). Green - ab108198 observed at 105 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab108198 was shown to specifically react with alpha Actinin 4 in wild-type HAP1 cells as signal was lost in ACTN4 (alpha Actinin 4) knockout cells. Wild-type and ACTN4 (alpha Actinin 4) knockout samples were subjected to SDS-PAGE. ab108198 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 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.
Immunoprecipitation - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198)

Purified ab108198 at 1/50 dilution (2µg) immunoprecipitating alpha Actinin 4 in HeLa whole cell lysate.
Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10µg
Lane 2 (+): ab108198 + HeLa whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab108198 in HeLa whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 105 kDa
Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198)

Flow cytometry protocols
Immunoprecipitation protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

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Publishing research using ab108198? Please let us know so that we can cite the reference in this datasheet.

ab108198 has been referenced in 18 publications.

Engevik AC et al. Recruitment of Polarity Complexes and Tight Junction Proteins to the Site of Apical Bulk Endocytosis. Cell Mol Gastroenterol Hepatol 12:59-80 (2021). PubMed: 33548596
Alday-Parejo B et al. MAGI1 localizes to mature focal adhesion and modulates endothelial cell adhesion, migration and angiogenesis. Cell Adh Migr 15:126-139 (2021). PubMed: 33823745
Furmanik M et al. Endoplasmic Reticulum Stress Mediates Vascular Smooth Muscle Cell Calcification via Increased Release of Grp78 (Glucose-Regulated Protein, 78 kDa)-Loaded Extracellular Vesicles. Arterioscler Thromb Vasc Biol 41:898-914 (2021). PubMed: 33297752
Zhang K et al. Nuclear exosome HMGB3 secreted by nasopharyngeal carcinoma cells promotes tumour metastasis by inducing angiogenesis. Cell Death Dis 12:554 (2021). PubMed: 34050127
Maier JI et al. EPB41L5 controls podocyte extracellular matrix assembly by adhesome-dependent force transmission. Cell Rep 34:108883 (2021). PubMed: 33761352

View all Publications for this product

Customer reviews and Q&As

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1-9 of 9 Abreviews or Q&A

Application

Immunocytochemistry/ Immunofluorescence

Sample

Human Cell (U2OS)

Permeabilization

Yes - NP40

Specification

U2OS

Blocking step

BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C

Fixative

Formaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Feb 22 2021

Application

Immunocytochemistry/ Immunofluorescence

Sample

Mouse Cell (Mouse vascular smooth muscle cells)

Permeabilization

Yes - NP40

Specification

Mouse vascular smooth muscle cells

Blocking step

BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C

Fixative

Formaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Feb 22 2021

Application

Western blot

Sample

Mouse Cell lysate - whole cell (Mouse vascular smooth muscle cells)

Gel Running Conditions

Reduced Denaturing

Loading amount

30 µg

Specification

Mouse vascular smooth muscle cells

Blocking step

Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Dec 09 2020

Application

Immunohistochemistry (Frozen sections)

Sample

Mouse Tissue sections (Hippocampus (Neurons in the CA1 Region))

Permeabilization

Yes - 0,1% Triton in PBS

Specification

Hippocampus (Neurons in the CA1 Region)

Blocking step

Serum as blocking agent for 45 minute(s) · Concentration: 10% · Temperature: 25°C

Fixative

Paraformaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Helena Lichtenfeld

Verified customer

Submitted Aug 21 2020

Application

Immunocytochemistry

Sample

Human Cell (HeLa cells)

Permeabilization

Yes - 0.5% Triton-X

Specification

HeLa cells

Blocking step

BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 25°C

Fixative

Formaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Anugraha Rajagopalan

Verified customer

Submitted Oct 29 2018

Application

Western blot

Sample

Human Cell lysate - whole cell (Vascular smooth muscle cells)

Gel Running Conditions

Reduced Denaturing

Loading amount

25 µg

Specification

Vascular smooth muscle cells

Blocking step

Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Dec 02 2016

Application

Western blot

Sample

Rat Cell lysate - other (Primary lung epithelia cells)

Gel Running Conditions

Reduced Denaturing (4-12% Nupage)

Specification

Primary lung epithelia cells

Blocking step

(agent) for 2 hour(s) and 0 minute(s) · Concentration: 0.2% · Temperature: 22°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Nov 02 2015

Application

Western blot

Sample

Human Cell lysate - whole cell (Breast)

Loading amount

20 µg

Specification

Breast

Gel Running Conditions

Reduced Denaturing (10%)

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Aug 15 2012

Application

Immunocytochemistry/ Immunofluorescence

Sample

Human Cell (Breast)

Specification

Breast

Fixative

Formaldehyde

Permeabilization

Yes - 0.2% Triton X-100

Blocking step

Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Aug 15 2012

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Recombinant Protein

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Involvement in disease

Sequence similarities

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

References (18)

Customer reviews and Q&As

Immunocytochemistry/ Immunofluorescence abreview for Anti-alpha Actinin 4 antibody [EPR2533(2)]

Immunocytochemistry/ Immunofluorescence abreview for Anti-alpha Actinin 4 antibody [EPR2533(2)]

Western blot abreview for Anti-alpha Actinin 4 antibody [EPR2533(2)]

Immunohistochemistry (Frozen sections) abreview for Anti-alpha Actinin 4 antibody [EPR2533(2)]

Immunocytochemistry abreview for Anti-alpha Actinin 4 antibody [EPR2533(2)]

Western blot abreview for Anti-alpha Actinin 4 antibody [EPR2533(2)]

Western blot abreview for Anti-alpha Actinin 4 antibody [EPR2533(2)]

Western blot abreview for Anti-alpha Actinin 4 antibody [EPR2533(2)]

Immunocytochemistry/ Immunofluorescence abreview for Anti-alpha Actinin 4 antibody [EPR2533(2)]