Anti-Pin1 Antibody Picoband™ (monoclonal, 5E5)

Product Name

Anti-Pin1 Antibody Picoband™ (monoclonal, 5E5)

View all PIN1 Antibodies

SKU/Catalog Number

M00467-1

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-Pin1 Antibody Picoband™ (monoclonal, 5E5) catalog # M00467-1. Tested in Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human, Monkey, Rat.

Storage & Handling

At -20°C for one year from date of receipt. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for six months. Avoid repeated freezing and thawing.

Cite This Product

Anti-Pin1 Antibody Picoband™ (monoclonal, 5E5) (Boster Biological Technology, Pleasanton CA, USA, Catalog # M00467-1)

Host

Mouse

Contents

Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.

Clonality

Monoclonal

Clone Number

5E5

Isotype

IgG2b

Immunogen

E.coli-derived human Pin1 recombinant protein (Position: M1-E163). Human Pin1 shares 95% amino acid (aa) sequence identity with mouse Pin1.

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins.

Reactive Species

M00467-1 is reactive to PIN1 in Human, Monkey, Rat

Applications

M00467-1 is guaranteed for Flow Cytometry, IF, ICC, WB Boster Guarantee

Observed Molecular Weight

18 kDa

Calculated molecular weight

18.243kDa

Background of PIN1

Peptidyl-prolyl cis-trans isomerase NIMA-interacting 1, also called DOD, is an enzyme that in humans is encoded by the PIN1 gene. It is mapped to 19p13.2. The enzyme binds to a subset of proteins and thus plays a role as a post phospho PIN1rol in regulating protein function. Studies have shown that the deregulation of PIN1 may play a pivotal role in various diseases. Notably, the up-regulation of PIN1 may be implicated in certain cancers, and the down-regulation of Pin1 may be implicated in Alzheimer's disease. Inhibitors of PIN1 may have therapeutic implications for cancer and immune disorders. PIN1 activity regulates the outcome of proline-directed kinase (e.g. MAPK, CDK or GSK3) signalling and consequently regulates cell proliferation (in part through control of cyclin D1 levels and stability) and cell survival. PIN1 also has an essential role in maintaining cell proliferation and regulating cyclin D1 function.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconsitution

Adding 0.2 ml of distilled water will yield a concentration of 500 µg/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Western blot, 0.25-0.5 µg/ml, Human, Monkey, Rat
Immunocytochemistry/Immunofluorescence, 5 µg/ml, Human
Flow Cytometry, 1-3 µg/1x10⁶ cells, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of Pin1 using anti-Pin1 antibody (M00467-1).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human HEK293 whole cell lysates,
Lane 3: human SW620 whole cell lysates,
Lane 4: monkey COS-7 whole cell lysates,
Lane 5: rat brain tissue lysates,
Lane 6: rat C6 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-Pin1 antigen affinity purified monoclonal antibody (Catalog # M00467-1) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1001) with Tanon 5200 system. A specific band was detected for Pin1 at approximately 18 kDa. The expected band size for Pin1 is at 18 kDa.

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Figure 2. IF analysis of Pin1 using anti-Pin1 antibody (M00467-1).
Pin1 was detected in an immunocytochemical section of Hela cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL mouse anti-Pin1 Antibody (M00467-1) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

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Figure 3. Flow Cytometry analysis of U87 cells using anti-Pin1 antibody (M00467-1).
Overlay histogram showing U87 cells stained with M00467-1 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-Pin1 Antibody (M00467-1, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Gene/Protein Information For PIN1 (Source: Uniprot.org, NCBI)

Alternative Names

dod; EC 5.2.1.8; peptidylprolyl cis/trans isomerase, NIMA-interacting 1; peptidyl-prolyl cis/trans isomerase, NIMA-interacting; peptidyl-prolyl cis-trans isomerase NIMA-interacting 1; Peptidyl-prolyl cis-trans isomerase Pin1; Pin1; PPIase Pin1; prolyl isomerase; protein (peptidyl-prolyl cis/trans isomerase) NIMA-interacting 1; protein (peptidylprolyl cis/trans isomerase) NIMA-interacting 1; Rotamase Pin1; UBL5 PIN1 DOD, UBL5 peptidylprolyl cis/trans isomerase, NIMA-interacting 1 peptidyl-prolyl cis-trans isomerase NIMA-interacting 1|PPIase Pin1|protein (peptidyl-prolyl cis/trans isomerase) NIMA-interacting 1|protein interacting with never in mitosis A1|rotamase Pin1

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