Anti-PCNA Rabbit Monoclonal Antibody

Product Name

Anti-PCNA Rabbit Monoclonal Antibody

View all PCNA Antibodies

SKU/Catalog Number

M00125-1

Size

100 μl

Form

Liquid

Description

Boster Bio Anti-PCNA Rabbit Monoclonal Antibody catalog # M00125-1. Tested in WB, IHC, ICC/IF, IP, Flow Cytometry applications. This antibody reacts with Human, Mouse, Rat.

Storage & Handling

Store at -20°C for one year. For short term storage and frequent use, store at 4°C for up to one month. Avoid repeated freeze-thaw cycles.

Cite This Product

Anti-PCNA Rabbit Monoclonal Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # M00125-1)

Host

Rabbit

Contents

Rabbit IgG in phosphate buffered saline, pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol, 0.4-0.5mg/ml BSA.

Clonality

Monoclonal

Clone Number

DO-16

Isotype

Rabbit IgG

Immunogen

A synthesized peptide derived from human PCNA

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

Reactive Species

M00125-1 is reactive to PCNA in Human, Mouse, Rat

Applications

M00125-1 is guaranteed for Flow Cytometry, IP, IF, IHC, ICC, WB Boster Guarantee

Observed Molecular Weight

36 kDa

Calculated molecular weight

28.769kDa

Background of PCNA

Phosphoinositide-3-kinase (PI3K) that phosphorylates PtdIns (Phosphatidylinositol), PtdIns4P (Phosphatidylinositol 4-phosphate) and PtdIns (4,5) P2 (Phosphatidylinositol 4,5-bisphosphate) to generate phosphatidylinositol 3,4,5-trisphosphate (PIP3) . PIP3 plays a key role by recruiting PH domain-containing proteins to the membrane, including AKT1 and PDPK1, activating signaling cascades involved in cell growth, survival, proliferation, motility and morphology. Participates in cellular signaling in response to various growth factors.

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

Restore with deionized water (or equivalent) for reconstitution volume of 1.0 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.

WB 1:3000-1:10000
IHC 1:50-1:200
ICC/IF 1:100-1:500
IP 1:50-1:100
FC 1:200-1:500

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of PCNA using anti-PCNA antibody (M00125-1).
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 HepG2 whole cell lysates,
Lane 3: human U-937 whole cell lysates,
Lane 4: human 293T whole cell lysates,
Lane 5: mouse spleen tissue lysates,
Lane 6: rat RAW264.7 tissue 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 rabbit anti-PCNA antigen affinity purified monoclonal antibody (Catalog # M00125-1) at 1:3000 overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for PCNA at approximately 36 kDa. The expected band size for PCNA is at 29 kDa.

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Immunohistochemical analysis of paraffin-embedded human colon, using PCNA Antibody.

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Figure 3. IF analysis of PCNA using anti-PCNA antibody (M00125-1) and anti-Beta Tubulin antibody (M01857-3).
PCNA was detected in immunocytochemical section of U2OS cell. 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 at 1:100 with rabbit anti-PCNA Antibody (M00125-1) and mouse anti-Beta Tubulin antibody (M01857-3) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) and Dylight550-conjugated Anti- mouse IgG Secondary Antibody (red)(Catalog#BA1133) were used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

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Immunoprecipitate (IP) analysis using the Antibody at 1:50 dilution. (wb at 1:3K dilution)

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

Alternative Names

cyclin; DNA polymerase delta auxiliary protein; MGC8367; proliferating cell nuclear antigen PCNA ATLD2 proliferating cell nuclear antigen proliferating cell nuclear antigen|DNA polymerase delta auxiliary protein|cyclin

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