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PAP (Prostatic Acid Phophatase), Affinity Purified Antibody, Chicken

MBS555374

0.2 mL

430 USD

Antibody

PAP (Prostatic Acid Phophatase), Affinity Purified Antibody, Chicken

[PAP (Prostatic Acid Phophatase)]

[prostatic acid phosphatase isoform PAP; Prostatic acid phosphatase; prostatic acid phosphatase; acid phosphatase, prostate; 5'-nucleotidase (EC:3.1.3.5); 5'-NT]

[PAP]

[ACPP; ACPP; ACP3; 5'-NT; ACP-3; PAP; 5'-NT; TMPase]

PPAP_HUMAN

pab

Chicken

Mouse

386

Liquid. PBS (50% by volume) and glycerol (50% by volume) supplemented with 0.02% sodium azide as a preservative. Concentration: 10 mg/ml of chicken total IgY antibody supplemented with 20 ug/ml of affinity purified antibody.

Store at 4°C in the dark. Under these conditions, the antibody should have a shelf life of at least 12 months (provided they remain sterile). Do not freeze the antibody unless you want to store it for longer periods of time. Note: each time an antibody preparation is frozen, about half its binding activity is lost.

Immunohistochemistry (IHC)

Immunohistochemistry: 1:500-1:1,000. Dilutions listed as a recommendation. Optimal dilution should be determined by investigator. Immunostaining Cell Cultures . 1. Draw of culture medium with aspirator and add 1 ml of 3.7 % formalin in PBS solution to the dish. (make up from 10mls Fisher 37% formalin plus 90mls PBS, the Fisher formalin contains 37% formaldehyde plus about 1% methanol which may be relevant sometimes). Let sit at room temp for 1 minute. (can add 0.1% Tween 20 to PBS used here and all subsequent steps to reduce background; probably best not to do this first time round though as it may extract your antigen or help wash your cells off the dish). 2. Take off the formalin/PBS and add 1ml of cold methanol (-20°C, kept in well sealed bottle in fridge). Let sit for no more than 1 minute. 3. Take off methanol and add 1ml of PBS, not letting the specimen dry out. To block nonspecific antibody binding can add ~10ml (=1%) of goat serum (Sigma), and can incubate for 30 minutes. Can then add antibody reagents. Typically 100ml of hybridoma tissue culture supernatent or 1ml of mouse ascites fluid or crude serum. Incubate for 1 hour at room temp. (or can go at 37°C for 30 minutes to 1 hour, or can do 4°C overnight, exact time not too critical). Can do very gentle shaking for well adherent cell lines (3T3, Hek293 etc.). 4. Remove primary antibody and replace with 1 ml of PBS. Let sit for 5-10 minutes, replace PBS and repeat twice, to give three washes in PBS. 5. Add 0.5 mls of secondary antibody. These are fluorescently labeled Goat anti mouse or rabbit antibodies and are conjugated to ALEXA dyes and are from Molecular probes (Eugene Oregon, the ALEXA dyes are sulphonated rhodamine compounds and are much more stable to UV than FITC, TRITC, Texas red etc.). Typically make 1:2,000 dilutions of these secondaries in PBS plus 1% goat serum, BSA or non fat milk carrier. Incubate for 1 hour at room temp. (or can go at 37°C for 30 minutes to 1 hour, or can do 4°C overnight). Can do gentle shaking for well adherent cell lines (3T3, HEK293 etc.). 6. Remove secondary antibody and replace with 1 ml of PBS. Let sit for 5-10 minutes, replace PBS and repeat twice, to give three washes in PBS. 7. Drop on one drop of Fisher mounting medium onto dish and apply 22mm square coverslip. View in the microscope!

Testing Data

Type Info: Chicken IgY

Immunogen: Recombinant mouse PAP protein was expressed using a baculoviral-delivery system. Immunostaining Tissue: Solutions . PBS - sodium phosphate-buffered (100 mM; pH 7.2) isotonic (0.9% NaCl, w/v) saline Antibody dilution buffer (PBS with 0.1% non-ionic detergent, such as Triton X-100 or Tween-20). For anti-fading, use Catalog#: MBS555421 or make your won fluorescein anti-fading reagent -- Make up a 2 mg/ml phenylene diamine solution in PBS (phenylene diamine requires extensive vortexing to put it into solution). Once the phenylene diamine is completely dissolved, add an equal volume of glycerol and mix. This reagent will last about a week at -20 degree C. Discard this reagent when it starts to turn dark brown. Other Reagents . Fluorescein-labeled goat anti-chicken IgY. 1. Prepare your tissue sections or cultured cells as you normally would. Wash your sections or cells for 1 min with PBS at room temperature. 2. Incubate your sections or cells with your chicken primary antibodies (diluted in "antibody dilution buffer") for at least 1 hour at room temperature. The concentration of your antibody may be anywhere from 1:50-1:150 depending on the titre of the antibody and the concentration of your antigen. 3. Wash your sections or cells over a 10 minute period at room temperature (with two changes of PBS). 4. Incubate your sections or cells with fluorescein-labeled goat anti-chicken IgY (1:500 dilution in "antibody dilution buffer” for 1 hour at room temperature. Be sure to keep these slides or culture dishes in subdued light (e.g., in a drawer) to avoid bleaching of the fluorescein dye. 5. Repeat step #4. 6. Add a drop of "fluorescence anti-fading reagent" (i-BRITE Plus) to your sections or cells. Place a coverslip over the section. If you want to reduce messiness, you may also seal the coverslip by painting the edges with nail polish. 7. Store the slides or culture dishes in the refrigerator (in the dark).

Antibodies; Pain and Inflammation; PAP (Prostatic Acid Phosphatase)

Mouse PAP is a 43,698 dalton protein (381 amino acids; NCBI accession numberAAF23171) associated with prostatic cancer cells, as well as primary afferent sensory neurons involved in the pain pathway. This protein is an enzyme that dephosphorylates adenosine monophosphate (AMP) in the dorsal horn gray matter of the spinal cord, generating free adenosine. Injections of PAP into the dorsal horn of experimental mice has been shown to decrease pain perception by acting in an antinociceptive, antihyperalgesic, and antiallodynic fashion.

6382064

NP_001090.2

NM_001099.4

40,443 Da

This gene encodes an enzyme that catalyzes the conversion of orthophosphoric monoester to alcohol and orthophosphate. It is synthesized under androgen regulation and is secreted by the epithelial cells of the prostate gland. An alternatively spliced transcript variant encoding a longer isoform has been found for this gene. This isoform contains a transmembrane domain and is localized in the plasma membrane-endosomal-lysosomal pathway. [provided by RefSeq, Sep 2008]

ACPP: A non-specific tyrosine phosphatase that dephosphorylates a diverse number of substrates under acidic conditions (pH 4-6) including alkyl, aryl, and acyl orthophosphate monoesters and phosphorylated proteins. Has lipid phosphatase activity and inactivates lysophosphatidic acid in seminal plasma. Belongs to the histidine acid phosphatase family. 2 isoforms of the human protein are produced by alternative splicing. Protein type: EC 3.1.3.2; Phosphatase; Cofactor and Vitamin Metabolism - riboflavin; EC 3.1.3.5; Motility/polarity/chemotaxis. Chromosomal Location of Human Ortholog: 3q22.1. Cellular Component: multivesicular body; extracellular space; Golgi cisterna; lysosomal membrane; apical part of cell; integral to membrane; plasma membrane; intracellular; vesicle membrane; nucleus; secretory granule; filopodium. Molecular Function: 5 -nucleotidase activity; choline binding; identical protein binding; acid phosphatase activity; protein homodimerization activity; phosphoric monoester hydrolase activity; thiamin phosphate phosphatase activity. Biological Process: thiamin metabolic process; dephosphorylation; positive regulation of adenosine receptor signaling pathway; nucleotide metabolic process; regulation of sensory perception of pain; adenosine metabolic process; purine base metabolic process

0.2 mL

430 USD