Recombinant Anti-CYP11A1 antibody [EPR24868-86] (ab272494)
![Western blot - Anti-CYP11A1 antibody [EPR24868-86] (ab272494)](https://www.abcam.com/ps/products/272/ab272494/Images/ab272494-480575-anti-cyp11a1-antibody-epr2486886-western-blot-tissue.jpg "Western blot - Anti-CYP11A1 antibody [EPR24868-86] (ab272494)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR24868-86] to CYP11A1
- Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra)
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-CYP11A1 antibody [EPR24868-86]
See all CYP11A1 primary antibodies -
Description
Rabbit monoclonal [EPR24868-86] to CYP11A1 -
Host species
Rabbit -
Specificity
ICC application does not react with Mouse and Rat species.
Flow Cyt application does not react with Mouse species.
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Tested applications
Suitable for: WB, IHC-P, ICC/IF, IP, 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.
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Positive control
- WB: Human adrenal gland, Human testis, JEG-3, Mouse testis, Rat testis, Mouse ovary, Rat ovary, NIH/3T3, PC-12 and Rat-1 lysates. IHC-P: Human adrenal gland, Human testis, Mouse testis, Rat adrenal gland tissues. ICC/IF: JEG-3 cells. Flow Cyt: JEG-3 and PC-12 cells. IP: JEG-3, Mouse testis and Rat testis 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. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
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
EPR24868-86 -
Isotype
IgG -
Research areas
- Metabolism
- Pathways and Processes
- Metabolic signaling pathways
- Lipid and lipoprotein metabolism
- Lipid metabolism
- Metabolism
- Pathways and Processes
- Metabolic signaling pathways
- Lipid and lipoprotein metabolism
- Cholesterol Metabolism
Associated products
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Compatible Secondaries
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab272494 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 |
1/1000. Predicted molecular weight: 60 kDa.
|
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| IHC-P |
1/10000.
|
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| ICC/IF |
1/50.
ICC application does not react with Mouse and Rat species. |
|
| IP |
1/30.
|
|
| Flow Cyt (Intra) |
1/50.
FC application does not react with Mouse species.
|
| Notes |
|---|
|
WB
1/1000. Predicted molecular weight: 60 kDa. |
|
IHC-P
1/10000. |
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ICC/IF
1/50. ICC application does not react with Mouse and Rat species. |
|
IP
1/30. |
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Flow Cyt (Intra)
1/50. FC application does not react with Mouse species.
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Target
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Function
Catalyzes the side-chain cleavage reaction of cholesterol to pregnenolone. -
Pathway
Lipid metabolism; C21-steroid hormone metabolism. -
Involvement in disease
Defects in CYP11A1 are a cause of congenital adrenal insufficiency (CAI).
Defects in CYP11A1 are a cause of congenital lipoid adrenal hyperplasia (CLAH) [MIM:201710]; also known as lipoid CAH. CLAH is the most severe form of adrenal hyperplasia. This autosomal recessive and potentially lethal condition includes the onset of profound adrenocortical insufficiency shortly after birth, hyperpigmentation reflecting increased production of pro-opiomelanocortin, elevated plasma renin activity as a consequence of reduced aldosterone synthesis, and male pseudohermaphroditism resulting from deficient fetal testicular testosterone synthesis. CLAH is a rare disease, except in Japan and Korea where it accounts for a significant percentage of cases of congenital adrenal hyperplasia. -
Sequence similarities
Belongs to the cytochrome P450 family. -
Cellular localization
Mitochondrion membrane. - Information by UniProt
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Database links
- Entrez Gene: 1583 Human
- Entrez Gene: 13070 Mouse
- Entrez Gene: 29680 Rat
- Omim: 118485 Human
- SwissProt: P05108 Human
- SwissProt: Q9QZ82 Mouse
- SwissProt: P14137 Rat
- Unigene: 303980 Human
see all -
Alternative names
- Cholesterol 20 22 desmolase antibody
- Cholesterol desmolase antibody
- Cholesterol monooxygenase (side chain cleaving) antibody
see all
Images
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Western blot - Anti-CYP11A1 antibody [EPR24868-86] (ab272494)All lanes : Anti-CYP11A1 antibody [EPR24868-86] (ab272494) at 1/1000 dilution
Lane 1 : Human adrenal gland tissue lysate
Lane 2 : Human testis tissue lysate
Lane 3 : Human heart tissue lysate
Lane 4 : JEG-3 (Human placenta epithelial choriocarcinoma) whole cell 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: 60 kDa
Observed band size: 50 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST
Negative control: human heart (PMID:19491374, 21520051)
The band below (~37 kDa) in lane 2 may be caused by degradation.
Exposure time: Lane 1: 7.75 seconds Lane 2-4: 37 seconds
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CYP11A1 antibody (ab272494)Immunohistochemical analysis of paraffin-embedded Human adrenal gland tissue labelling CYP11A1 with ab272494 at 1/10000 (0.054 ug/ml) followed by a ready to use LeicaDS9800 (BNOD™, Polymer Refine Detection) was used. Positive staining on human adrenal cortex (PMID: 22796438). (A) Low‑powered (magnification, x40) and (B) high‑powered (magnification, x400) microscopic images. The section was incubated with ab272494 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) was used.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 minsn was used.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CYP11A1 antibody (ab272494)Immunohistochemical analysis of paraffin-embedded Human testis tissue labelling CYP11A1 with ab272494 at 1/10000 (0.054 ug/ml) followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Positive staining on Leydig cells of human testis. The section was incubated with ab272494 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) was used.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
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![Immunocytochemistry/ Immunofluorescence - Anti-CYP11A1 antibody [EPR24868-86] (ab272494)](https://www.abcam.com/ps/products/272/ab272494/Images/ab272494-6-anti-cyp11a1-antibody-epr2486886-immunocytochemistry-jeg3.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-CYP11A1 antibody [EPR24868-86] (ab272494)Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized JEG-3 (human placenta epithelial choriocarcinoma) cells labelling CYP11A1 with ab272494 at 1/50 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining colocalized with mitochondrial marker (ab33985) in JEG-3 cell line is observed. ab33985 Anti-COX IV mouse monoclonal antibody - Mitochondrial Marker was used to counterstain mitochondrion at 1/1000 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.
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Flow Cytometry (Intracellular) - Anti-CYP11A1 antibody (ab272494)Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized JEG-3 (Human placenta epithelial choriocarcinoma) cells labelling CYP11A1 with ab272494 at 1/500 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, ab150081) at 1/2000 dilution was used as the secondary antibody.
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Immunoprecipitation - Anti-CYP11A1 antibody (ab272494)CYP11A1 was immunoprecipitated from 0.35 mg JEG-3 (Human placenta epithelial choriocarcinoma) whole cell lysate with ab272494 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab272494 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: JEG-3 (Human placenta epithelial choriocarcinoma) whole cell lysate 10µg
Lane 2: ab272494 IP in JEG-3 whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab272494 in JEG-3 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 58 seconds
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![Western blot - Anti-CYP11A1 antibody [EPR24868-86] (ab272494)](https://www.abcam.com/ps/products/272/ab272494/Images/ab272494-480576-anti-cyp11a1-antibody-epr2486886-western-blot-lysate.jpg)
Western blot - Anti-CYP11A1 antibody [EPR24868-86] (ab272494)All lanes : Anti-CYP11A1 antibody [EPR24868-86] (ab272494) at 1/1000 dilution
Lane 1 : Mouse testis tissue lysate
Lane 2 : Mouse brain tissue lysate
Lane 3 : Rat testis tissue lysate
Lane 4 : Rat brain tissue lysate
Lane 5 : Mouse ovary tissue lysate
Lane 6 : Rat ovary tissue lysate
Lane 7 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate
Lane 8 : PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate
Lane 9 : Rat-1 (rat embryonic fibroblast) 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: 60 kDa
Observed band size: 50 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST
Low expression: brain (PMID: 21520051, 19491374)
Lane 7-9: This blot was developed using a higher sensitivity ECL substrate.
The band below (~37 kDa) in lane 1 may be caused by degradation.
Exposure time: Lane 1-4, 7-9: 3 minutes Lane 5-6: 37 seconds
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CYP11A1 antibody (ab272494)Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labelling CYP11A1 with ab272494 at 1/10000 (0.054 ug/ml) followed by a ready to use LeicaDS9800(BOND™ Polymer Refine Detection) was used. Positive staining on Leydig cells of mouse testis. The section was incubated with ab272494 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) was used.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
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Flow Cytometry (Intracellular) - Anti-CYP11A1 antibody (ab272494)Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized PC-12 (Rat adrenal gland pheochromocytoma) cells labelling CYP11A1 with ab272494 at 1/50 dilution (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, ab150081) at 1/2000 dilution was used as the secondary antibody.
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Immunoprecipitation - Anti-CYP11A1 antibody (ab272494)CYP11A1 was immunoprecipitated from 0.35 mg Mouse testis tissue lysate with ab272494 at 1/30 dilution (2 μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab272494 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: Mouse testis tissue lysate 10µg
Lane 2: ab272494 IP in Mouse testis tissue lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab272494 in mouse testis tissue lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 10 seconds
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CYP11A1 antibody (ab272494)Immunohistochemical analysis of paraffin-embedded Rat adrenal gland tissue labelling CYP11A1 with ab272494 at 1/10000 (0.054 ug/ml) followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection)was used. Positive staining on rat adrenal cortex. The section was incubated with ab272494 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) was used.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
-
Immunoprecipitation - Anti-CYP11A1 antibody (ab272494)CYP11A1 was immunoprecipitated from 0.35 mg Rat testis tissue lysate with ab272494 at 1/30 dilution (2 μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab272494 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: Rat testis tissue lysate 10µg
Lane 2: ab272494 IP in Rat testis tissue lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab272494 in rat testis tissue lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 58 seconds
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CYP11A1 antibody (ab272494)Immunohistochemical analysis of paraffin-embedded Human cardiac muscle tissue labelling CYP11A1 with ab272494 at 1/10000 (0.054 ug/ml) followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Negative control: no staining on human cardiac muscle. The section was incubated with ab272494 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)was used.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) 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)
ab272494 has not yet been referenced specifically in any publications.