Recombinant Anti-CYP11A1 antibody [EPR24868-86] - BSA and Azide free (ab288573)
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 - BSA and Azide free
- Suitable for: IHC-P, mIHC, ICC/IF, WB, 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] - BSA and Azide free
See all CYP11A1 primary antibodies -
Description
Rabbit monoclonal [EPR24868-86] to CYP11A1 - BSA and Azide free -
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: IHC-P, mIHC, ICC/IF, WB, 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 and testis tissue lysates; Mouse testis and ovary tissue lysates; Rat testis and ovary tissue lysates; JERG-3, NIH/3T3, PC-12 and Rat-1 whole cell lysates. IHC-P: Human adrenal gland and testis, Mouse testis tissue; Rat adrenal gland tissue. ICC: JEG-3 cells. Flow Cyt: JEG-3 and PC-12 cells. IP: JEG-3 while cell lysate; Mouse testis tissue lysate; Rat testis tissue lysate. mIHC: Human testis.
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General notes
ab288573 is the carrier-free version of ab272494
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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. -
Storage buffer
pH: 7.2
Constituent: 100% PBS -
Carrier free
Yes -
Concentration information loading...
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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
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab288573 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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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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mIHC |
Use at an assay dependent concentration.
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ICC/IF |
Use at an assay dependent concentration.
ICC application does not react with Mouse and Rat species. |
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WB |
Use at an assay dependent concentration. Detects a band of approximately 50 kDa (predicted molecular weight: 60 kDa).
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IP |
Use at an assay dependent concentration.
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
FC application does not react with Mouse species. |
Notes |
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IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
mIHC
Use at an assay dependent concentration. |
ICC/IF
Use at an assay dependent concentration. ICC application does not react with Mouse and Rat species. |
WB
Use at an assay dependent concentration. Detects a band of approximately 50 kDa (predicted molecular weight: 60 kDa). |
IP
Use at an assay dependent concentration. |
Flow Cyt (Intra)
Use at an assay dependent concentration. FC application does not react with Mouse species. |
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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Multiplex immunohistochemistry - Anti-CYP11A1 antibody [EPR24868-86] - BSA and Azide free (ab288573)
Fluorescence multiplex immunohistochemical analysis of human adrenal gland (formalin-fixed paraffin-embedded section). Panel A shows merged staining of anti-PCSK2 stained on adrenal medulla (ab274418; gray; Opal™690) at 1:2000 (0.263 μg/ml) [Panel B], anti-CYP11A1 stained on adrenal cortex (ab272494; green; Opal™520) at 1:10000 (0.053 μg/ml) [Panel C], and anti-Collagen VI stained on extracellular matrix (ab182744; red; Opal™570) at 1:1000 (1.518 μg/ml) [Panel D] on human adrenal gland. DAPI was used as a nuclear counter stain. Followed by Opal Polymer HRP Ms + Rb secondary antibody. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope. The section was incubated in three rounds of staining: in the order of ab274418, ab272494, and ab182744 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins was used.
This data was developed using ab272494, the same antibody clone in a different buffer formulation.
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Multiplex immunohistochemistry - Anti-CYP11A1 antibody [EPR24868-86] - BSA and Azide free (ab288573)
Fluorescence multiplex immunohistochemical analysis of human adrenal gland (formalin-fixed paraffin-embedded section). Panel A shows merged staining of anti-PCSK2 stained on adrenal medulla (ab274418; gray; Opal™690) at 1:2000 (0.263 μg/ml) [Panel B], anti-CYP11A1 stained on adrenal cortex (ab272494; green; Opal™520) at 1:10000 (0.053 μg/ml) [Panel C], and anti-Neuropeptide Y stained on chromaffin cells (ab221145; red; Opal™570) at 1:2000 (0.279 μg/ml) [Panel D] on human adrenal gland. DAPI was used as a nuclear counter stain. Followed by Opal Polymer HRP Ms + Rb secondary antibody. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope. The section was incubated in three rounds of staining: in the order of ab274418, ab272494, and ab221145 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins was used.
This data was developed using ab272494, the same antibody clone in a different buffer formulation.
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Multiplex immunohistochemistry - Anti-CYP11A1 antibody [EPR24868-86] - BSA and Azide free (ab288573)
This data was developed using ab272494, the same antibody clone in a different buffer formulation.
Multiplex immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human testis tissue.
Panel A: Merged staining of anti-Vimentin (ab193555; red; Opal™690), anti-CYP11A1 (ab272494; cyan; Opal™520) and anti-DDX4 / MVH (ab270534; green; Opal™570) on human testis.
Panel B: Anti-CYP11A1 stained on Leydig cells.
Panel C: Anti-DDX4 / MVH stained on all spermatogenic cell types.
Panel D: Anti-Vimentin stained on Sertoli cells and fibroblasts.
Key protocol steps: The section was incubated in three rounds of staining: in the order of ab193555 (1:2000 dilution) and ab272494 (1:10000 dilution) for 30 mins, then ab270534 (1:2000 dlilution) for 10 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.
DAPI was used as a nuclear counter stain. Opal Polymer HRP Ms + Rb was used as a secondary.
Antigen retrieval: Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
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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?This data was developed using ab272494, the same antibody clone in a different buffer formulation.
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 times: Lane 1: 7.75 seconds; Lane 2-4: 37 seconds.
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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?This data was developed using ab272494, the same antibody clone in a different buffer formulation.
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 times: 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 [EPR24868-86] - BSA and Azide free (ab288573)
This data was developed using ab272494, the same antibody clone in a different buffer formulation.
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 (Bond™ Polymer Refine Detection). 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).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CYP11A1 antibody [EPR24868-86] - BSA and Azide free (ab288573)
This data was developed using ab272494, the same antibody clone in a different buffer formulation.
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).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CYP11A1 antibody [EPR24868-86] - BSA and Azide free (ab288573)
This data was developed using ab272494, the same antibody clone in a different buffer formulation.
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). 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).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CYP11A1 antibody [EPR24868-86] - BSA and Azide free (ab288573)
This data was developed using ab272494, the same antibody clone in a different buffer formulation.
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). 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).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CYP11A1 antibody [EPR24868-86] - BSA and Azide free (ab288573)
This data was developed using ab272494, the same antibody clone in a different buffer formulation.
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).
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] - BSA and Azide free (ab288573)
This data was developed using ab272494, the same antibody clone in a different buffer formulation.
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. 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).
-ve control 1: ab272494 at 1/50 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 dilution.
-ve control 2: ab33985 at 1/1000 dilution, followed by 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 [EPR24868-86] - BSA and Azide free (ab288573)
This data was developed using ab272494, the same antibody clone in a different buffer formulation.
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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Flow Cytometry (Intracellular) - Anti-CYP11A1 antibody [EPR24868-86] - BSA and Azide free (ab288573)
This data was developed using ab272494, the same antibody clone in a different buffer formulation.
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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This data was developed using ab272494, the same antibody clone in a different buffer formulation.
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 10 ug
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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This data was developed using ab272494, the same antibody clone in a different buffer formulation.
CYP11A1 was immunoprecipitated from 0.35 mg Mouse testis tissue 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: Mouse testis tissue lysate 10 ug
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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This data was developed using ab272494, the same antibody clone in a different buffer formulation.
CYP11A1 was immunoprecipitated from 0.35 mg Rat testis tissue 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: Rat testis tissue lysate 10 ug
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
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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Datasheet download
Certificate of Compliance
References (0)
ab288573 has not yet been referenced specifically in any publications.