Anti-hnRNP E2/PCBP2 Antibody Picoband™

Product Name

Anti-hnRNP E2/PCBP2 Antibody Picoband™

View all PCBP2 Antibodies

SKU/Catalog Number

A02425-3

Size

100 µg/vial

Form

Lyophilized

Description

Boster Bio Anti-hnRNP E2/PCBP2 Antibody Picoband™ catalog # A02425-3. Tested in ELISA, Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human, Mouse, 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-hnRNP E2/PCBP2 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A02425-3)

Host

Rabbit

Contents

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

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

E.coli-derived human hnRNP E2/PCBP2 recombinant protein (Position: D136-K276).

*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

A02425-3 is reactive to PCBP2 in Human, Mouse, Rat

Applications

A02425-3 is guaranteed for ELISA, Flow Cytometry, IF, ICC, WB Boster Guarantee

Observed Molecular Weight

40 kDa

Calculated molecular weight

38.58kDa

Background of PCBP2

Poly(rC)-binding protein 2 is a protein that in humans is encoded by the PCBP2 gene. The protein encoded by this gene appears to be multifunctional. Along with PCBP-1 and hnRNPK, it is one of the major cellular poly(rC)-binding proteins. The encoded protein contains three K-homologous (KH) domains which may be involved in RNA binding. Together with PCBP-1, this protein also functions as a translational coactivator of poliovirus RNA via a sequence-specific interaction with stem-loop IV of the IRES, promoting poliovirus RNA replication by binding to its 5'-terminal cloverleaf structure. It has also been implicated in translational control of the 15-lipoxygenase mRNA, human papillomavirus type 16 L2 mRNA, and hepatitis A virus RNA. The encoded protein is also suggested to play a part in formation of a sequence-specific alpha-globin mRNP complex which is associated with alpha-globin mRNA stability. This multiexon structural mRNA is thought to be retrotransposed to generate PCBP-1, an intronless gene with functions similar to that of PCBP2. This gene and PCBP-1 have paralogous genes (PCBP3 and PCBP4) which are thought to have arisen as a result of duplication events of entire genes. This gene also has two processed pseudogenes (PCBP2P1 and PCBP2P2). Multiple transcript variants encoding different isoforms have been found for this gene.

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, 0.1-0.25 µg/ml, Human, Mouse, Rat
Immunocytochemistry/Immunofluorescence, 5 µg/ml, Human
Flow Cytometry, 1-3 µg/1x10⁶ cells, Human
Direct ELISA, 0.1-0.5 µg/ml, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of hnRNP E2/PCBP2 using anti-hnRNP E2/PCBP2 antibody (A02425-3).
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 Jurkat whole cell lysates,
Lane 3: human 293T whole cell lysates,
Lane 4: human Raji whole cell lysates,
Lane 5: rat PC-12 whole cell lysates,
Lane 6: rat C6 whole cell lysates,
Lane 7: mouse RAW264.7 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 rabbit anti-hnRNP E2/PCBP2 antigen affinity purified polyclonal antibody (Catalog # A02425-3) at 0.25 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-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 hnRNP E2/PCBP2 at approximately 40 kDa. The expected band size for hnRNP E2/PCBP2 is at 39 kDa.

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Figure 2. IF analysis of hnRNP E2/PCBP2 using anti-hnRNP E2/PCBP2 antibody (A02425-3) and anti-Tubulin Alpha antibody (M03989-3).
hnRNP E2/PCBP2 was detected in immunocytochemical section of PC-3 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 with 5 μg/mL rabbit anti-hnRNP E2/PCBP2 Antibody (A02425-3) and mouse anti-Tubulin Alpha antibody (M03989-3) overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) and DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) 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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Figure 3. Flow Cytometry analysis of Hela cells using anti-hnRNP E2/PCBP2 antibody (A02425-3).
Overlay histogram showing Hela cells stained with A02425-3 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-hnRNP E2/PCBP2 Antibody (A02425-3, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Gene/Protein Information For PCBP2 (Source: Uniprot.org, NCBI)

Alternative Names

alpha-CP2; Heterogeneous nuclear ribonucleoprotein E2; heterogenous nuclear ribonucleoprotein E2; hnRNP E2; hnRNP-E2; HNRPE2; MGC110998; poly(rC) binding protein 2; poly(rC)-binding protein 2 PCBP2 HNRNPE2, HNRPE2, hnRNP-E2 poly(rC) binding protein 2 poly(rC)-binding protein 2|alpha-CP2|epididymis secretory sperm binding protein|heterogeneous nuclear ribonucleoprotein E2|heterogenous nuclear ribonucleoprotein E2|hnRNP E2

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