Anti-PGP9.5 Antibody Picoband™ (monoclonal, 3E4)

Product Name

Anti-PGP9.5 Antibody Picoband™ (monoclonal, 3E4)

View all UCH-L1/PGP9.5 Antibodies

SKU/Catalog Number

M01018-6

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-PGP9.5 Antibody Picoband™ (monoclonal, 3E4) catalog # M01018-6. Tested in Flow Cytometry, IHC, WB applications. This antibody reacts with Human, Mouse, Rat.

Storage & Handling

Store 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 freeze-thaw cycles.

Cite This Product

Anti-PGP9.5 Antibody Picoband™ (monoclonal, 3E4) (Boster Biological Technology, Pleasanton CA, USA, Catalog # M01018-6)

Host

Mouse

Contents

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

Clonality

Monoclonal

Clone Number

3E4

Isotype

Mouse IgG1

Immunogen

A synthetic peptide corresponding to a sequence at the C-terminus of human PGP9.5, different from the related mouse and rat sequences by two amino acids.

*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

M01018-6 is reactive to UCHL1 in Human, Mouse, Rat

Applications

M01018-6 is guaranteed for Flow Cytometry, IHC, WB Boster Guarantee

Observed Molecular Weight

27 kDa

Calculated molecular weight

24.824kDa

Background of UCH-L1/PGP9.5

UCH-L1, also known as PGP9.5, is a member of a gene family whose products hydrolyze small C-terminal adducts of ubiquitin to generate the ubiquitin monomer. Expression of UCH-L1 is highly specific to neurons and to cells of the diffuse neuroendocrine system and their tumors. It is abundantly present in all neurons (accounts for 1-2% of total brain protein), expressed specifically in neurons and testis/ovary. The catalytic triad of UCH-L1 contains a cysteine at position 90, an aspartate at position 176, and a histidine at position 161 that are responsible for its hydrolase activity.

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

Add 0.2ml of distilled water will yield a concentration of 500ug/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, 0.25-0.5μg/ml, Human, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Human, Rat
Flow Cytometry, 1-3μg/1x10⁶ cells, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of PGP9.5 using anti-PGP9.5 antibody (M01018-6).
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 30ug of sample under reducing conditions.
Lane 1: human U87 whole cell lysates,
Lane 2: human SH-SY5Y whole cell lysates,
Lane 3: rat brain tissue lysates,
Lane 4: mouse brain tissue lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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-PGP9.5 antigen affinity purified monoclonal antibody (Catalog # M01018-6) 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 PGP9.5 at approximately 27KD. The expected band size for PGP9.5 is at 27KD.

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Figure 2. IHC analysis of PGP9.5 using anti-PGP9.5 antibody (M01018-6).
PGP9.5 was detected in paraffin-embedded section of human renal clear cell carcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml mouse anti-PGP9.5 Antibody (M01018-6) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.

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Figure 3. IHC analysis of PGP9.5 using anti-PGP9.5 antibody (M01018-6).
PGP9.5 was detected in paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml mouse anti-PGP9.5 Antibody (M01018-6) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.

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Figure 4. Flow Cytometry analysis of 293T cells using anti-PGP9.5 antibody (M01018-6).
Overlay histogram showing 293T cells stained with M01018-6 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-PGP9.5 Antibody (M01018-6, 1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Gene/Protein Information For UCHL1 (Source: Uniprot.org, NCBI)

Alternative Names

EC 3.4.19.12; EC 6.-; Neuron cytoplasmic protein 9.5; PARK5; PGP 9.5; PGP9.5; PGP9.5Uch-L1; PGP95; ubiquitin carboxyl-terminal esterase L1 (ubiquitin thiolesterase); ubiquitin carboxyl-terminal hydrolase isozyme L1; ubiquitin C-terminal hydrolase; Ubiquitin thioesterase L1; UCHL1; UCH-L1 UCHL1 HEL-117, HEL-S-53, NDGOA, PARK5, PGP 9.5, PGP9.5, PGP95, SPG79, Uch-L1 ubiquitin C-terminal hydrolase L1 ubiquitin carboxyl-terminal hydrolase isozyme L1|epididymis luminal protein 117|epididymis secretory protein Li 53|neuron cytoplasmic protein 9.5|ubiquitin carboxyl-terminal esterase L1 (ubiquitin thiolesterase)|ubiquitin thioesterase L1|ubiquitin thiolesterase

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