Aromatase Antibody - BSA Free

Novus Biologicals | Catalog # NB100-1596

Novus Biologicals
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Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse, Rat, Primate, Rabbit

Cited:

Human, Mouse, Rat, Primate, Rabbit

Predicted:

Bovine (90%), Equine (91%). Backed by our 100% Guarantee.

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Immunocytochemistry/ Immunofluorescence, Simple Western

Cited:

Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Immunocytochemistry/ Immunofluorescence, Simple Western, IF/IHC

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
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Product Specifications

Immunogen

A synthetic peptide made to a C-terminal portion of the human Aromatase protein (between residues 400-502). [UniProt# P11511]

Reactivity Notes

Use in Rabbit reported in scientific literature (PMID:35245530). Rabbit reactivity reported in scientific literature (PMID: 25850953). Immunogen displays the following percentage of sequence identity for non-tested species: horse (91%), cow (90%), pig (83%), sheep (81%), goat (81%), and dog (81%) proteins. Rat reactivity reported in scientific literature (PMID: 29898754). Use in Mouse reported in scientific literature (PMID:32634520).

Localization

Membrane-bound

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

55 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for Aromatase Antibody - BSA Free

Western Blot: Aromatase Antibody [NB100-1596]

Western Blot: Aromatase Antibody [NB100-1596]

Aromatase-Antibody-Western-Blot-NB100-1596-img0005.jpg
Immunocytochemistry/ Immunofluorescence: Aromatase Antibody [NB100-1596]

Immunocytochemistry/ Immunofluorescence: Aromatase Antibody [NB100-1596]

Immunocytochemistry/Immunofluorescence: Aromatase Antibody [NB100-1596] - U2OS cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti-Aromatase Antibody NB100-1596 at 2 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.
Immunohistochemistry-Paraffin: Aromatase Antibody [NB100-1596]

Immunohistochemistry-Paraffin: Aromatase Antibody [NB100-1596]

Aromatase-Antibody-Immunohistochemistry-Paraffin-NB100-1596-img0006.jpg
Western Blot: Aromatase Antibody [NB100-1596]

Western Blot: Aromatase Antibody [NB100-1596]

Western Blot: Aromatase Antibody [NB100-1596] - Detection of aromatase in human fetal temporal lobe lysate using NB 100-1596. ECL exposure, 1-2 minutes.
Immunohistochemistry-Paraffin: Aromatase Antibody [NB100-1596]

Immunohistochemistry-Paraffin: Aromatase Antibody [NB100-1596]

Aromatase-Antibody-Immunohistochemistry-Paraffin-NB100-1596-img0007.jpg
Simple Western: Aromatase Antibody [NB100-1596]

Simple Western: Aromatase Antibody [NB100-1596]

Aromatase Antibody-Simple Western-NB100-1596-img0009.jpg
Aromatase Antibody

Immunohistochemistry: Aromatase Antibody [NB100-1596] -

Immunohistochemistry: Aromatase Antibody [NB100-1596] - Expression of the cytochrome P450 aromatase in the ovary of control (C; open bar; n = 6) & hypothyroid (Hypo; solid bar; n = 6) rabbits. Representative immunoblot showing the expression of aromatase (a) & Ponceau's Red stained membrane (b). Relative expression of aromatase in C & Hypo groups (c). Data are mean ± SEM. ∗∗P < 0.01. Immunoreactivity of antiaromatase (+) was identified in granulosa cells of antral follicles (tertiary, tf; Graafian, gf) of control (d, e) & hypothyroid (f, g) ovaries. Nonlabeling was observed when the primary antibody was omitted (negative control; h). Bars: (d) 200 μm; (f) & (h) 100 μm; & (e) & (g) 20 μm. Granulose cells, gc; theca cells, tc. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/28133606), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Aromatase Antibody

Immunohistochemistry: Aromatase Antibody [NB100-1596] -

Immunohistochemistry: Aromatase Antibody [NB100-1596] - Expression of the cytochrome P450 aromatase in the ovary of control (C; open bar; n = 6) & hypothyroid (Hypo; solid bar; n = 6) rabbits. Representative immunoblot showing the expression of aromatase (a) & Ponceau's Red stained membrane (b). Relative expression of aromatase in C & Hypo groups (c). Data are mean ± SEM. ∗∗P < 0.01. Immunoreactivity of antiaromatase (+) was identified in granulosa cells of antral follicles (tertiary, tf; Graafian, gf) of control (d, e) & hypothyroid (f, g) ovaries. Nonlabeling was observed when the primary antibody was omitted (negative control; h). Bars: (d) 200 μm; (f) & (h) 100 μm; & (e) & (g) 20 μm. Granulose cells, gc; theca cells, tc. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/28133606), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Aromatase Antibody

Immunohistochemistry: Aromatase Antibody [NB100-1596] -

Immunohistochemistry: Aromatase Antibody [NB100-1596] - Expression of the cytochrome P450 aromatase in the ovary of control (C; open bar; n = 6) & hypothyroid (Hypo; solid bar; n = 6) rabbits. Representative immunoblot showing the expression of aromatase (a) & Ponceau's Red stained membrane (b). Relative expression of aromatase in C & Hypo groups (c). Data are mean ± SEM. ∗∗P < 0.01. Immunoreactivity of antiaromatase (+) was identified in granulosa cells of antral follicles (tertiary, tf; Graafian, gf) of control (d, e) & hypothyroid (f, g) ovaries. Nonlabeling was observed when the primary antibody was omitted (negative control; h). Bars: (d) 200 μm; (f) & (h) 100 μm; & (e) & (g) 20 μm. Granulose cells, gc; theca cells, tc. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/28133606), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Aromatase Antibody

Immunohistochemistry: Aromatase Antibody [NB100-1596] -

Immunohistochemistry: Aromatase Antibody [NB100-1596] - Expression of the cytochrome P450 aromatase in the ovary of control (C; open bar; n = 6) & hypothyroid (Hypo; solid bar; n = 6) rabbits. Representative immunoblot showing the expression of aromatase (a) & Ponceau's Red stained membrane (b). Relative expression of aromatase in C & Hypo groups (c). Data are mean ± SEM. ∗∗P < 0.01. Immunoreactivity of antiaromatase (+) was identified in granulosa cells of antral follicles (tertiary, tf; Graafian, gf) of control (d, e) & hypothyroid (f, g) ovaries. Nonlabeling was observed when the primary antibody was omitted (negative control; h). Bars: (d) 200 μm; (f) & (h) 100 μm; & (e) & (g) 20 μm. Granulose cells, gc; theca cells, tc. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/28133606), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Aromatase Antibody

Immunohistochemistry: Aromatase Antibody [NB100-1596] -

Immunohistochemistry: Aromatase Antibody [NB100-1596] - Expression of the cytochrome P450 aromatase in the ovary of control (C; open bar; n = 6) & hypothyroid (Hypo; solid bar; n = 6) rabbits. Representative immunoblot showing the expression of aromatase (a) & Ponceau's Red stained membrane (b). Relative expression of aromatase in C & Hypo groups (c). Data are mean ± SEM. ∗∗P < 0.01. Immunoreactivity of antiaromatase (+) was identified in granulosa cells of antral follicles (tertiary, tf; Graafian, gf) of control (d, e) & hypothyroid (f, g) ovaries. Nonlabeling was observed when the primary antibody was omitted (negative control; h). Bars: (d) 200 μm; (f) & (h) 100 μm; & (e) & (g) 20 μm. Granulose cells, gc; theca cells, tc. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/28133606), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Aromatase Antibody

Simple Western: Aromatase Antibody [NB100-1596] -

Simple Western: Aromatase Antibody [NB100-1596] - Metformin decreases aromatase-positive, tumor-associated macrophages. a Quantification of aromatase-positive cells & b CD68-positive (CD68+) macrophages costaining for aromatase in the tumor border of tissues from control or metformin-treated rats. c Representative control & metformin-treated IHC images of CD68 & aromatase dual staining in tissue bordering mammary tumors (T) (arrows: yellow = CD68+aromatase+, brown = CD68+; scale bar = 50um). d Aromatase expression by Western blotting (WES system) from in vitro activated M1 & M2 rat macrophages using two different primary antibodies (Ab #1: Novus NB100-1596; Ab #2: clone 677, Baylor College of Medicine). Controls include ovary (positive control) & ovary in which the primary antibody was preincubated with an aromatase blocking peptide for 1 h, as well as KGN cells with (positive control) & without (negative control) forskolin treatment. e IHC intensity score for ER & PR (0 = no stain; 1 = weak; 2 = moderate; 3 = strong staining). *p < 0.05 Image collected & cropped by CiteAb from the following publication (https://breast-cancer-research.biomedcentral.com/articles/10.1186/s1305…), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for Aromatase Antibody - BSA Free

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:100

Immunohistochemistry

1:200. Use reported in scientific literature (PMID 28133606)

Immunohistochemistry-Paraffin

1:200

Simple Western

reported in scientific literature (PMID 29898754)

Western Blot

1 ug/ml
Application Notes
In Western blot, a band is seen at ~55 kDa in human brain, and Immunocytochemistry/Immunofluorescence where membrane staining is observed in SH-SY-5Y cells. The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: Aromatase

Aromatase (ARO1) or cytochrome P450 19A1 (CYP19A1) belongs to cytochrome P450 family of enzymes, which are monooxygenases known for their essential role in drug catabolism and cholesterol/steroid synthesis. Employing heme-group act as a co-factor, ARO1 catalyzes aromatic C18 estrogens formation from C19 androgens and is responsible for conversion of testosterone to estradiol and androstenedione to estrone. ARO1 is localized to cellular membranes as a peripheral membrane protein and in addition to female reproductive tract and adipose tissue, aromatase can be found in testis, liver, brain as well as certain tumors including breast cancer. During ontogenesis, it plays an important role in the development of CNS and gonads as well as sex differentiation. Defective ARO1 may leads to aromatase excess syndrome (AEXS) and aromatase deficiency (AROD). Moreover, obesity has been linked to abnormally high ARO1 expression in mammary tissues, increased local estrogen production, and susceptibility to breast hyperplasia and cancer. ARO1 is the key enzyme in intratumoral estrogen production in postmenopausal mammary cancer.

Long Name

Aromatase

Alternate Names

ARO1, CYAR, CYP19, CYP19A1, CYPXIX, EC 1.14.14.14, Estrogen synthase

Gene Symbol

CYP19A1

Additional Aromatase Products

Product Documents for Aromatase Antibody - BSA Free

Certificate of Analysis

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Product Specific Notices for Aromatase Antibody - BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Citations for Aromatase Antibody - BSA Free

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Protocols

View specific protocols for Aromatase Antibody - BSA Free (NB100-1596):

Immunocytochemistry Protocol

Culture cells to appropriate density in 35 mm culture dishes or 6-well plates.

1. Remove culture medium and wash the cells briefly in PBS. Add 10% formalin to the dish and fix at room temperature for 10 minutes.
2. Remove the formalin and wash the cells in PBS.
3. Permeablize the cells with 0.1% Triton X100 or other suitable detergent for 10 min.
4. Remove the permeablization buffer and wash three times for 10 minutes each in PBS. Be sure to not let the specimen dry out.
5. To block nonspecific antibody binding, incubate in 10% normal goat serum from 1 hour to overnight at room temperature.
6. Add primary antibody at appropriate dilution and incubate overnight at 4C.
7. Remove primary antibody and replace with PBS. Wash three times for 10 minutes each.
8. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
9. Remove secondary antibody and replace with PBS. Wash three times for 10 minutes each.
10. Counter stain DNA with DAPi if required.

Immunohistochemistry-Paraffin Embedded Sections

Antigen Unmasking:
Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes (keep slides in the sodium citrate buffer at all times).

Staining:
1. Wash sections in deionized water three times for 5 minutes each.
2. Wash sections in PBS for 5 minutes.
3. Block each section with 100-400 ul blocking solution (1% BSA in PBS) for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul diluted primary antibody. Incubate overnight at 4 C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul HRP polymer conjugated secondary antibody. Incubate 30 minutes at room temperature.
7. Wash sections three times in wash buffer for 5 minutes each.
8. Add 100-400 ul DAB substrate to each section and monitor staining closely.
9. As soon as the sections develop, immerse slides in deionized water.
10. Counterstain sections in hematoxylin.
11. Wash sections in deionized water two times for 5 minutes each.
12. Dehydrate sections.
13. Mount coverslips.


Western Blot Protocol

1. Perform SDS-PAGE on samples to be analyzed, loading 10-25 ug of total protein per lane.
2. Transfer proteins to PVDF membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
3. Stain the membrane with Ponceau S (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
4. Rinse the blot TBS -0.05% Tween 20 (TBST).
5. Block the membrane in 5% Non-fat milk in TBST (blocking buffer) for at least 1 hour.
6. Wash the membrane in TBST three times for 10 minutes each.
7. Dilute primary antibody in blocking buffer and incubate overnight at 4C with gentle rocking.
8. Wash the membrane in TBST three times for 10 minutes each.
9. Incubate the membrane in diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturer's instructions) for 1 hour at room temperature.
10. Wash the blot in TBST three times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturer's instructions.

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for Aromatase Antibody - BSA Free

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  • Q: I have a customer with a question about antibody NB100-1596 anti-aromatase. The customer is questioning why the recommended storage conditions are 4C with no freezing. They are concerned that the antibody will not be stable unless it is frozen.

    A: For this product, we found that the antibody was quite unstable after just one freeze thaw cycle, which is why we recommend storage at 4C. In our experience, the antibody performs quite well when stored at 4C; even for long periods of time. I hope this will help your customer. Please let them know that if they freeze the antibody, we will not be able to guarantee function.

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