PRODUCT CODE: ER1901-84

NAPSIN A Rabbit Polyclonal Antibody (ER1901-84)

Applications

  • WB

  • ICC

  • IHC-P

  • FC

REACTIVITY

  • Human

Western blot analysis of NAPSIN A on human lung tissue lysate. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody was used at a 1:500 dilution in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
  • Western blot analysis of NAPSIN A on human lung tissue lysate. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody was used at a 1:500 dilution in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
  • ICC staining of NAPSIN A in 293T cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ER1901-84,  1/100 dilution) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).
  • Immunohistochemical analysis of paraffin-embedded human lung cancer tissue using anti-NAPSIN A antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ER1901-84,  1/200 dilution) for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. Counter stained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-NAPSIN A antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ER1901-84,  1/200 dilution) for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. Counter stained with hematoxylin and mounted with DPX.
  • Flow cytometric analysis of NAPSIN A was done on A549 cells. The cells were fixed, permeabilized and stained with the primary antibody (ER1901-84, 1/100) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated goat anti-rabbit IgG Secondary antibody at 1/500 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Western blot analysis of NAPSIN A on human lung tissue lysate. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody was used at a 1:500 dilution in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.

Applications

  • WB

  • ICC

  • IHC-P

  • FC

REACTIVITY

  • Human

SPECIFICATIONS

Product Type

Rabbit polyclonal primary

Product Name

NAPSIN A Rabbit Polyclonal Antibody (ER1901-84)

Immunogen

Recombinant protein within human napsin a aa 44-183.

Host

Rabbit

Positive Control

Human lung tissue, 293T, human lung cancer tissue, human kidney tissue.

Conjugation

Unconjugated

Clonality

Polyclonal

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Protein affinity purified.

MOLECULAR WEIGHT

Predicted band size 11/30/45 kDa.

Isotype

IgG

APPLICATION DILUTION

  • WB:1:500-1:1,000

  • ICC:1:50-1:200

  • IHC-P:1:50-1:200

  • FC:1:50-1:100

TARGET

UNIPROT #

PROTEIN NAME

NAPSIN A

SYNONYMS

Asp 4 antibody; ASP4 antibody; Aspartyl protease 4 antibody; KAP antibody; Kdap antibody; Kidney derived aspartic protease like protein antibody; NAP1 antibody; NAPA antibody; Napsa antibody; NAPSA_HUMAN antibody; Napsin 1 antibody; napsin A aspartic peptidase antibody; Napsin A precursor antibody; Napsin-1 antibody; Napsin-A antibody; Pronapsin A antibody; SNAPA antibody; TA01/TA02 antibody

SEQUENCE SIMILARITIES

Belongs to the peptidase A1 family.

TISSUE SPECIFICITY

Expressed predominantly in adult lung (type II pneumocytes) and kidney and in fetal lung. Low levels in adult spleen and very low levels in peripheral blood leukocytes.

SUBCELLULAR LOCATION

Secreted.

FUNCTION

Napsin A is an asparatic protease with a molecular weight of approximately 38 kDa, expressed in type-II pneumocytes and is involved in the N- and C-terminal processing of proSP-B in type-II pneumocytes. TAO2 has been shown to be identical with Napsin A. There is also a Napsin B gene which is transcribed exclusively in cells related to the immune system but lacks a stop codon and may represent a transcribed pseudogene. It is expressed in the cytoplasm and is strongly positive in up to 80% of primary lung adenocarcinomas by immunohistochemistry. Poorly differentiated cancers do not stain as well as those that are well-differentiated. Squamous cell carcinomas and small cell carcinomas of the lung have been negative for napsin A. However, 10% of renal cell carcinomas and thyroid carcinomas are also positive. Renal and thyroid cancers may give false positive results, most likely because of the presence of intrinsic biotin, which can be detected on negative controls. Less than 5% of assorted adenocarcinomas, including those from the breast, pancreas, biliary tract, and colon stain with napsin A: expression, when present in breast and colonic adenocarcinomas, appears to be granular, unlike that in the lung.