Recombinant Anti-ATP1A1+ATP1A2+ATP1A3+ATP1A4 antibody [EPR25414-249] (ab300507)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR25414-249] to ATP1A3 + alpha 1 Sodium Potassium ATPase + ATP1A2 + ATP1A4
- Suitable for: ICC/IF, WB, IHC-P, IHC-Fr, Flow Cyt (Intra)
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
-
Product name
Anti-ATP1A1+ATP1A2+ATP1A3+ATP1A4 antibody [EPR25414-249] -
Description
Rabbit monoclonal [EPR25414-249] to ATP1A3 + alpha 1 Sodium Potassium ATPase + ATP1A2 + ATP1A4 -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, WB, IHC-P, IHC-Fr, Flow Cyt (Intra)more details
Unsuitable for: IP -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
-
Positive control
- WB: A-172, SH-SY5Y, U-87 MG, Neuro-2a whole cell lysates; HEK-293 transfected with ATP1A1-A4 (WT) expression vector containing a myc-His-tag®, whole cell lysate; Human cerebellum, Mouse brain, Rat brain tissue lysates. IHC-P: Human cardiac muscle, Human cerebrum, Mouse cerebrum, Rat cardiac muscle, Rat cerebrum FFPE tissue sections. IHCFr: Mouse + Rat cerebrum frozen tissue. ICC/IF: HEK293T, A-172, Neuro-2a cell lines. Flow Cyt (Intra): A-172 (human brain glioblastoma), Neuro-2a (Mouse neuroblastoma).
-
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
-
Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
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
EPR25414-249 -
Isotype
IgG -
Research areas
Associated products
-
Alternative Versions
-
Compatible Secondaries
-
Isotype control
-
Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab300507 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 |
---|---|---|
ICC/IF |
1/50.
|
|
WB |
1/1000. Detects a band of approximately 110 kDa (predicted molecular weight: 111 kDa).
|
|
IHC-P |
1/5000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
|
|
IHC-Fr |
1/500.
|
|
Flow Cyt (Intra) |
1/500.
|
Notes |
---|
ICC/IF
1/50. |
WB
1/1000. Detects a band of approximately 110 kDa (predicted molecular weight: 111 kDa). |
IHC-P
1/5000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
IHC-Fr
1/500. |
Flow Cyt (Intra)
1/500. |
Target
-
Cellular localization
ATP1A3: Cell membrane. alpha 1 Sodium Potassium ATPase: Cell membrane. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV. ATP1A2: Membrane. Cell membrane. ATP1A4: Cell membrane. In mature sperm, located in the principle piece of the sperm flagellum. -
Database links
- Entrez Gene: 476 Human
- Entrez Gene: 477 Human
- Entrez Gene: 478 Human
- Entrez Gene: 480 Human
- Entrez Gene: 11928 Mouse
- Entrez Gene: 232975 Mouse
- Entrez Gene: 98660 Mouse
- Entrez Gene: 24211 Rat
see all
Images
-
All lanes : Anti-ATP1A1+ATP1A2+ATP1A3+ATP1A4 antibody [EPR25414-249] (ab300507) at 1/1000 dilution
Lane 1 : A-172 (human brain glioblastoma ) whole cell lysate
Lane 2 : SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate
Lane 3 : U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 111 kDa
Observed band size: 110 kDa why is the actual band size different from the predicted?
Exposure time: 48 secondsBlocking / Diluting buffer and concentration:5% NFDM/TBST
Note: The samples are unboiled
-
All lanes : Anti-ATP1A1+ATP1A2+ATP1A3+ATP1A4 antibody [EPR25414-249] (ab300507) at 1/5000 dilution
Lane 1 : Human cerebellum tissue lysate
Lane 2 : Mouse brain tissue lysate
Lane 3 : Rat brain tissue lysate
Lane 4 : Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 111 kDa
Observed band size: 110 kDa why is the actual band size different from the predicted?Blocking / Diluting buffer and concentration:5% NFDM/TBST.
Exposure time: Lanes 1-3: 3 seconds; lane 4: 15 seconds.
Note: The samples are unboiled.
-
Lane 1 : Anti-ATP1A1+ATP1A2+ATP1A3+ATP1A4 antibody [EPR25414-249] (ab300507) at 1/1000 dilution (HEK-293 (human embryonic kidney) transfected with an empty vector (vector control), containing a myc-His-tag®, whole cell lysate)
Lanes 2-5 : Anti-ATP1A1+ATP1A2+ATP1A3+ATP1A4 antibody [EPR25414-249] (ab300507) at 1/1000 dilution
Lane 2 : HEK-293 transfected with ATP1A1 (WT) expression vector containing a myc-His-tag®, whole cell lysate
Lane 3 : HEK-293 transfected with ATP1A2 (WT) expression vector containing a myc-His-tag®, whole cell lysate
Lane 4 : HEK-293 transfected with ATP1A3 (WT) expression vector containing a myc-His-tag®, whole cell lysate
Lane 5 : HEK-293 transfected with ATP1A4 (WT) expression vector containing a myc-His-tag®, whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 111 kDa
Observed band size: 110 kDa why is the actual band size different from the predicted?
Exposure time: 3 secondsBlocking / Diluting buffer and concentration:5% NFDM/TBST.
The samples are unboiled.
Note: Bands above 110 kDa are aggregates of Na+/K+-ATPase alpha subunits. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATP1A1+ATP1A2+ATP1A3+ATP1A4 antibody [EPR25414-249] (AB300507)
Immunohistochemical analysis of paraffin-embedded human cardiac muscle labeling ATP1A1+ATP1A2+ATP1A3+ATP1A4 with ab300507 at 1:5000 (0.108 μg/ml) followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection) kit. Positive staining on human cardiac muscle, particularly on the intercalated discs (PMID: 26109061). The section was incubated with ab300507 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection).
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATP1A1+ATP1A2+ATP1A3+ATP1A4 antibody [EPR25414-249] (AB300507)
Immunohistochemical analysis of paraffin-embedded human cerebrum labeling ATP1A1+ATP1A2+ATP1A3+ATP1A4 with ab300507 at 1:5000 (0.108 μg/ml) followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection) kit. Positive staining on human cerebrum. The section was incubated with ab300507 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection).
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATP1A1+ATP1A2+ATP1A3+ATP1A4 antibody [EPR25414-249] (AB300507)
Immunohistochemical analysis of paraffin-embedded mouse cerebrum labeling ATP1A1+ATP1A2+ATP1A3+ATP1A4 with ab300507 at 1:5000 (0.108 μg/ml) followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection) kit. Positive staining on mouse cerebrum (PMID: 32301109) . The section was incubated with ab300507 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection).
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATP1A1+ATP1A2+ATP1A3+ATP1A4 antibody [EPR25414-249] (AB300507)
Immunohistochemical analysis of paraffin-embedded rat cardiac muscle labeling ATP1A1+ATP1A2+ATP1A3+ATP1A4 with ab300507 at 1:5000 (0.108 μg/ml) followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection) kit. Positive staining on rat cardiac muscle, particularly on the intercalated discs. The section was incubated with ab300507 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection).
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATP1A1+ATP1A2+ATP1A3+ATP1A4 antibody [EPR25414-249] (AB300507)
Immunohistochemical analysis of paraffin-embedded rat cerebrum labeling ATP1A1+ATP1A2+ATP1A3+ATP1A4 with ab300507 at 1:5000 (0.108 μg/ml) followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection) kit. Positive staining on rat cerebrum. The section was incubated with ab300507 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection).
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins
-
Immunohistochemistry (Frozen sections) - Anti-ATP1A1+ATP1A2+ATP1A3+ATP1A4 antibody [EPR25414-249] (AB300507)
Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen sectioned mouse cerebrum (fresh) labeling ATP1A1+ATP1A2+ATP1A3+ATP1A4 with ab300507 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150081) at 1/1000 dilution (green). Positive staining on mouse cerebrum is observed. The nuclear counter stain is DAPI (blue).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150081) at 1/1000 dilution.
No antigen retrieval was carried out.
-
Immunohistochemistry (Frozen sections) - Anti-ATP1A1+ATP1A2+ATP1A3+ATP1A4 antibody [EPR25414-249] (AB300507)
Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen sectioned rat cerebrum (fresh) labeling ATP1A1+ATP1A2+ATP1A3+ATP1A4 with ab300507 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150081) at 1/1000 dilution (green). Positive staining on rat cerebrum is observed. The nuclear counter stain is DAPI (blue).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150081) at 1/1000 dilution.
No antigen retrieval was carried out.
-
Immunocytochemistry/ Immunofluorescence - Anti-ATP1A1+ATP1A2+ATP1A3+ATP1A4 antibody [EPR25414-249] (ab300507)
Immunofluorescent analysis of 4% Paraformaldehyde fixed, 0.1% Triton X-100 permeabilized HEK293T (human embryonic kidney epithelial cell) labelling ATP1A3 with ab300507 at 1/500 dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed secondary antibody at 1/1000 (2 μg/ml) dilution (green). Anti -Myc-Tag Mouse mAb (Alexa Fluor® 647) was used as a counterstain at 1/100 dilution (0.38μg/m) (red). Confocal image showing cytoplasmic and membranous staining in HEK-293T cells transfected with human ATP1A3 expression vector containing a myc tag. The nuclear counter stain is DAPI (blue).
-
Immunocytochemistry/ Immunofluorescence - Anti-ATP1A1+ATP1A2+ATP1A3+ATP1A4 antibody [EPR25414-249] (ab300507)
Immunofluorescent analysis of 4% Paraformaldehyde fixed, 0.1% Triton X-100 permeabilized HEK293T (human embryonic kidney epithelial cell) labelling ATP1A1+ATP1A2+ATP1A4 with ab300507 at 1/500 dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed secondary antibody at 1/1000 (2 μg/ml) dilution (green). Anti -Myc-Tag Mouse mAb (Alexa Fluor® 647) was used as a counterstain at 1/100 dilution (0.38μg/m) (red). Confocal image showing cytoplasmic and membranous staining in HEK-293T cells transfected with human ATP1A1 or ATP1A2 or ATP1A4 expression vector containing a myc tag. The nuclear counter stain is DAPI (blue).
-
Immunocytochemistry/ Immunofluorescence - Anti-ATP1A1+ATP1A2+ATP1A3+ATP1A4 antibody [EPR25414-249] (ab300507)
Immunofluorescent analysis of 100% methanol fixed, 0.1% Triton X-100 permeabilized A-172 (human brain glioblastoma) labelling ATP1A1+ATP1A2+ATP1A3+ATP1A4 with ab300507 at 1/50 dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed secondary antibody at 1/1000 (2 μg/ml) dilution (green). Confocal image showing membranous and weak cytoplasmic staining in A-172 cell line. The nuclear counter stain is DAPI (blue).
Tubulin is detected with ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) (red) at 1/200 dilution.
-
Immunocytochemistry/ Immunofluorescence - Anti-ATP1A1+ATP1A2+ATP1A3+ATP1A4 antibody [EPR25414-249] (ab300507)
Immunofluorescent analysis of 100% methanol fixed, 0.1% Triton X-100 permeabilized Neuro-2a (mouse neuroblastoma neuroblasts) labelling ATP1A1+ATP1A2+ATP1A3+ATP1A4 with ab300507 at 1/50 dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed secondary antibody at 1/1000 (2 μg/ml) dilution (green). Confocal image showing membrane and weak cytoplasm staining in Neuro-2a cell line. The nuclear counter stain is DAPI (blue).
Tubulin is detected with ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) (red) at 1/200 dilution.
-
Flow Cytometry (Intracellular) - Anti-ATP1A1+ATP1A2+ATP1A3+ATP1A4 antibody [EPR25414-249] (AB300507)
Flow Cytometry (Intracellular) analysis of A-172 (human brain glioblastoma) labeling ATP1A1+ATP1A2+ATP1A3+ATP1A4 (red) with ab300405 at a 1/500 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A Goat Anti-Rabbit IgG (Alexa Fluor® 488, (ab150081)) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730) Isotype control. Blue - (unlabeled control) - Cells without incubation with primary and secondary antibodies.
-
Flow Cytometry (Intracellular) - Anti-ATP1A1+ATP1A2+ATP1A3+ATP1A4 antibody [EPR25414-249] (AB300507)
Flow Cytometry (Intracellular) analysis of Neuro-2a (mouse neuroblastoma neuroblasts) labeling ATP1A1+ATP1A2+ATP1A3+ATP1A4 (red) with ab300405 at a 1/500 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A Goat Anti-Rabbit IgG (Alexa Fluor® 488, (ab150081)) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730) Isotype control. Blue - (unlabeled control) - Cells without incubation with primary and secondary antibodies.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
-
SDS download
-
Datasheet download
Certificate of Compliance
References (0)
ab300507 has not yet been referenced specifically in any publications.