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Key Product Details

Species Reactivity

Human, Mouse, Rat

Applications

Flow Cytometry, Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit

Format

BSA Free

Concentration

1.0 mg/ml

Product Summary for Aminopeptidase N/CD13 Antibody - BSA Free

Immunogen

Partial recombinant mouse CD13 protein (amino acids 69-966) [UniProt P97449]

Clonality

Polyclonal

Host

Rabbit

Scientific Data Images for Aminopeptidase N/CD13 Antibody - BSA Free

Western Blot: Aminopeptidase N/CD13 Antibody [NBP2-77451] - Total protein from human mouse and rat small intestine was separated on a 7.5% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 2.0 ug/ml anti-CD13 in blocking buffer and detected with an anti-rabbit HRP secondary antibody using West Pico PLUS chemiluminescence detection reagent.
Immunohistochemistry-Paraffin: Aminopeptidase N/CD13 Antibody [NBP2-77451] - IHC analysis of a formalin fixed paraffin embedded tissue section of human prostate cancer using CD13 antibody at 1:50 dilution. The signal was developed with HRP-DAB detection method and the nuclei were counterstained with hematoxylin staining. This CD13 antibody generated a specific membrane staining in the cancer cells and the staining was localized more towards the secretory surface of the glands.
Flow Cytometry: Aminopeptidase N/CD13 Antibody [NBP2-77451] - A surface stain was performed on THP-1 cells with CD13 Antibody NBP2-77451 (blue) and a matched isotype control (orange). Cells were incubated in an antibody dilution of 2.5 ug/mL for 20 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550.

Applications for Aminopeptidase N/CD13 Antibody - BSA Free

Application
Recommended Usage

Flow Cytometry

2-5 ug/ml

Immunohistochemistry

1:50 - 1:200

Immunohistochemistry-Paraffin

1:50 - 1:200

Western Blot

0.5 - 2.0 ug/ml
Please Note: Optimal dilutions of this antibody should be experimentally determined.

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 for up to 3 months. For longer storage, aliquot and store at -20C.

Background: Aminopeptidase N/CD13

CD13 (aminopeptidase N, APN) is a 150 kDa type II transmembrane zinc-binding ectopeptidase expressed on various cell types. This metalloprotease preferentially catalyzes removal of neutral amino acids from small peptides, thus activating or inactivating bioactive peptides. CD13 has also role in extracellular matrix degradation, antigen processing and signal transduction, is important in inflammatory responses, regulates intercellular contact, cell motility and vascularization. CD13 is involved in protection of leukemic cells against apoptosis and its expression associated with poor prognosis of carcinomas.

Alternate Names

Aminopeptidase M, ANPEP, APN, CD13, gp150, PEPN

Gene Symbol

ANPEP

Additional Aminopeptidase N/CD13 Products

Product Documents for Aminopeptidase N/CD13 Antibody - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for Aminopeptidase N/CD13 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.

Protocols

View specific protocols for Aminopeptidase N/CD13 Antibody - BSA Free (NBP2-77451):

Aminopeptidase N/CD13 Antibody:
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.

Staining:
1. Wash sections in deionized water three times for 5 minutes each.
2. Wash sections in wash buffer for 5 minutes.
3. Block each section with 100-400 ul blocking solution 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 biotinylated diluted secondary antibody. Incubate 30 minutes at room temperature.
7. Remove secondary antibody solution and wash sections three times with wash buffer for 5 minutes each.
8. Add 100-400 ul Streptavidin-HRP reagent to each section and incubate for 30 minutes at room temperature.
9. Wash sections three times in wash buffer for 5 minutes each.
10. Add 100-400 ul DAB substrate to each section and monitor staining closely.
11. As soon as the sections develop, immerse slides in deionized water.
12. Counterstain sections in hematoxylin.
13. Wash sections in deionized water two times for 5 minutes each.
14. Dehydrate sections.
15. Mount coverslips.
Aminopeptidase N/CD13 Antibody:
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 anti-CD13 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 manufacturers instructions.

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