Recombinant Anti-Sigma1-receptor antibody [EPR23266-69] (ab253192)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR23266-69] to Sigma1-receptor
- Suitable for: Flow Cyt (Intra), WB, IHC-P
- Knockout validated
- Reacts with: Rat, Human, Recombinant fragment
Related conjugates and formulations
Overview
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Product name
Anti-Sigma1-receptor antibody [EPR23266-69]
See all Sigma1-receptor primary antibodies -
Description
Rabbit monoclonal [EPR23266-69] to Sigma1-receptor -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-Pmore details
Unsuitable for: ICC/IF or IP -
Species reactivity
Reacts with: Rat, Human, Recombinant fragment -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Caco-2, A375, U-87 MG, HEK-293 and HeLa whole cell lysates; HEK-293T transfected with Sigma1-receptor whole cell lysate; Rat brain, hippocampus, lung and kidney tissue lysates. IHC-P: Human spinal cord and liver tissue; Rat spinal cord and liver tissue. Flow Cyt (intra): HT-29 cells.
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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
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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: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR23266-69 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab253192 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 |
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Flow Cyt (Intra) |
1/600.
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WB |
1/1000. Predicted molecular weight: 25 kDa.
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IHC-P |
1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Notes |
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Flow Cyt (Intra)
1/600. |
WB
1/1000. Predicted molecular weight: 25 kDa. |
IHC-P
1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Target
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Function
Functions in lipid transport from the endoplasmic reticulum and is involved in a wide array of cellular functions probably through regulation of the biogenesis of lipid microdomains at the plasma membrane. Involved in the regulation of different receptors it plays a role in BDNF signaling and EGF signaling. Also regulates ion channels like the potassium channel and could modulate neurotransmitter release. Plays a role in calcium signaling through modulation together with ANK2 of the ITP3R-dependent calcium efflux at the endoplasmic reticulum. Plays a role in several other cell functions including proliferation, survival and death. Originally identified for its ability to bind various psychoactive drugs it is involved in learning processes, memory and mood alteration. -
Tissue specificity
Widely expressed with higher expression in liver, colon, prostate, placenta, small intestine, heart and pancreas. Expressed in the retina by retinal pigment epithelial cells. -
Involvement in disease
Amyotrophic lateral sclerosis 16, juvenile -
Sequence similarities
Belongs to the ERG2 family. -
Cellular localization
Nucleus inner membrane. Nucleus outer membrane. Endoplasmic reticulum membrane. Lipid droplet. Cell junction. Cell membrane. Cell projection > growth cone. Targeted to lipid droplets, cholesterol and galactosylceramide-enriched domains of the endoplasmic reticulum. Enriched at cell-cell communication regions, growth cone and postsynaptic structures. Localization is modulated by ligand-binding. - Information by UniProt
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Database links
- Entrez Gene: 10280 Human
- Entrez Gene: 29336 Rat
- Omim: 601978 Human
- SwissProt: Q99720 Human
- SwissProt: Q9R0C9 Rat
- Unigene: 522087 Human
- Unigene: 1129 Rat
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Alternative names
- Aging associated gene 8 protein antibody
- Aging-associated gene 8 protein antibody
- AL024364 antibody
see all
Images
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All lanes : Anti-Sigma1-receptor antibody [EPR23266-69] (ab253192) at 1/5000 dilution
Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : SIGMAR1 knockout HEK293T cell lysate
Lane 3 : Wild-type HeLa cell lysate
Lane 4 : SIGMAR1 knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : goat anti-rabbit HRP at 0.2 µg/ml
Performed under reducing conditions.
Predicted band size: 25 kDaab253192 was shown to react with SIGMAR1 in wild-type HEK293T in Western blot with loss of signal observed in SIGMAR1 knockout cell line ab266619 (SIGMAR1 knockout cell lysate ab258666). Wild-type HEK293T and HeLa cell lysates and the corresponding SIGMAR1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab253192 overnight at 4 °C at a 1/5000 dilution. Blots were incubated with goat anti-rabbit HRP secondary antibodies at 0.2μg/mL before imaging.
These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.
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All lanes : Anti-Sigma1-receptor antibody [EPR23266-69] (ab253192) at 1/1000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : SIGMAR1 knockout HEK-293T cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : Caco-2 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 25 kDa
Observed band size: 24 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab253192 observed at 24 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
ab253192 was shown to react with Sigma1-receptor in wild-type HEK-293T cells in Western blot with loss of signal observed in SIGMAR1 knockout cell line ab266619 (SIGMAR1 knockout cell lysate ab258666). Wild-type HEK-293T and SIGMAR1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab253192 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 h at room temperature before imaging.
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Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HT-29 (Human colorectal adenocarcinoma epithelial cell) cells labelling Sigma1-receptor with ab253192 at 1/600 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sigma1-receptor antibody [EPR23266-69] (ab253192)
Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling Sigma1-receptor with ab253192 at 1/500 dilution (1.23 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat liver (PMID: 8954936). The section was incubated with ab253192 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins
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All lanes : Anti-Sigma1-receptor antibody [EPR23266-69] (ab253192) at 1/1000 dilution
Lane 1 : HEK-293T (human embryonic kidney) transfected with an empty vector (vector control), containing a myc-His-tag®, whole cell lysate
Lane 2 : HEK-293T transfected with Sigma1-receptor (WT) expression vector containing a myc-His-tag®, whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution
Predicted band size: 25 kDa
Observed band size: 25 kDaBlocking and diluting buffer and concentration: 5% NFDM/TBST
Exposure time: 3.25 seconds
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All lanes : Anti-Sigma1-receptor antibody [EPR23266-69] (ab253192) at 1/1000 dilution
Lane 1 : Rat brain tissue lysate
Lane 2 : Rat hippocampus tissue lysate
Lane 3 : Rat lung tissue lysate
Lane 4 : Rat kidney tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution
Predicted band size: 25 kDa
Observed band size: 25 kDaBlocking and diluting buffer and concentration: 5% NFDM/TBST
Exposure time: 3 minutes
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All lanes : Anti-Sigma1-receptor antibody [EPR23266-69] (ab253192) at 1/1000 dilution
Lane 1 : Caco-2 (human colorectal adenocarcinoma epithelial cell), whole cell lysate
Lane 2 : A375 (human malignant melanoma epithelial cell), whole cell lysate
Lane 3 : U-87 MG (human glioblastoma-astrocytoma epithelial cell), whole cell lysate
Lane 4 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
Lane 5 : 293T (human embryonic kidney epithelial cell), whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution
Predicted band size: 25 kDa
Observed band size: 25 kDaBlocking and diluting buffer and concentration: 5% NFDM/TBST
Exposure time: 3 minutes
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sigma1-receptor antibody [EPR23266-69] (ab253192)
Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling Sigma1-receptor with ab253192 at 1/500 dilution (1.23 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human liver (PMID: 8954936). The section was incubated with ab253192 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sigma1-receptor antibody [EPR23266-69] (ab253192)
Immunohistochemical analysis of paraffin-embedded Rat spinal cord tissue labeling Sigma1-receptor with ab253192 at 1/500 dilution (1.23 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat spinal cord (PMID: 23314020). The section was incubated with ab253192 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sigma1-receptor antibody [EPR23266-69] (ab253192)
Immunohistochemical analysis of paraffin-embedded Human spinal cord tissue labeling Sigma1-receptor with ab253192 at 1/500 dilution (1.23 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human spinal cord (PMID: 23314020). The section was incubated with ab253192 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins
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
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SDS download
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Datasheet download
Certificate of Compliance
References (0)
ab253192 has not yet been referenced specifically in any publications.