Recombinant Anti-CYP11A1 antibody [EPR24868-86] - BSA and Azide free (ab288573)

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.
Tested applications

Suitable for: IHC-P, 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.
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.
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
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.

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...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR24868-86
Isotype

IgG
Research areas

Compatible Secondaries
- VeriBlot for IP Detection Reagent (HRP) (ab131366)
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

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
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.
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.

Notes
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.
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.

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.
Target information above from: UniProt accession P05108 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
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
- Unigene: 302865 Mouse
- Unigene: 1401 Rat
see all
Alternative names
- Cholesterol 20 22 desmolase antibody
- Cholesterol desmolase antibody
- Cholesterol monooxygenase (side chain cleaving) antibody
- Cholesterol side chain cleavage enzyme antibody
- Cholesterol side chain cleavage enzyme mitochondrial antibody
- Cholesterol side-chain cleavage enzyme antibody
- CP11A_HUMAN antibody
- CYP11A antibody
- CYP11A1 antibody
- CYPXIA1 antibody
- Cytochrome P450 11A1 antibody
- Cytochrome P450 11A1 mitochondrial antibody
- Cytochrome P450 family 11 subfamily A polypeptide 1 antibody
- Cytochrome P450 subfamily XIA antibody
- Cytochrome P450(scc) antibody
- Cytochrome P450C11A1 antibody
- mitochondrial antibody
- P450SCC antibody
- Steroid 20 22 lyase antibody
see all

Western blot - Anti-CYP11A1 antibody [EPR24868-86] - BSA and Azide free (ab288573)

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.
Western blot - Anti-CYP11A1 antibody [EPR24868-86] - BSA and Azide free (ab288573)

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
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
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
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
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
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
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.
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.
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.
Immunoprecipitation - 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.

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
Immunoprecipitation - 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.

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
Immunoprecipitation - 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.

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
Anti-CYP11A1 antibody [EPR24868-86] - BSA and Azide free (ab288573)

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.

Click here to view the general protocols

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Overview

Product name

Description

Host species

Specificity

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Carrier free

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Compatible Secondaries

Isotype control

Applications

The Abpromise guarantee

Target

Function

Pathway

Involvement in disease

Sequence similarities

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

Datasheet download

Certificate of Compliance

References (0)

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