Anti-PSE/SPDEF Antibody Picoband™

Product Name

Anti-PSE/SPDEF Antibody Picoband™

View all SPDEF Antibodies

SKU/Catalog Number

A04625-1

Size

100 µg/vial

Form

Lyophilized

Description

Boster Bio Anti-PSE/SPDEF Antibody Picoband™ catalog # A04625-1. Tested in ELISA, Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human.

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-PSE/SPDEF Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A04625-1)

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 PSE/SPDEF recombinant protein (Position: E52-I335).

*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

A04625-1 is reactive to SPDEF in Human

Applications

A04625-1 is guaranteed for ELISA, Flow Cytometry, IF, ICC, WB Boster Guarantee

Observed Molecular Weight

44 kDa

Calculated molecular weight

37.518kDa

Background of SPDEF

SAM pointed domain-containing Ets transcription factor is a protein that in humans is encoded by the SPDEF gene. The protein encoded by this gene belongs to the ETS family of transcription factors. It is highly expressed in the prostate epithelial cells, and functions as an androgen-independent transactivator of prostate-specific antigen (PSA) promoter. Higher expression of this protein has also been reported in brain, breast, lung and ovarian tumors, compared to the corresponding normal tissues, and it shows better tumor-association than other cancer-associated molecules, making it a more suitable target for developing specific cancer therapies. Alternatively spliced 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, 0.25-0.5 µg/ml, Human
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 PSE/SPDEF using anti-PSE/SPDEF antibody (A04625-1).
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 MCF-7 whole cell lysates,
Lane 2: human PC-3 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-PSE/SPDEF antigen affinity purified polyclonal antibody (Catalog # A04625-1) 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-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 PSE/SPDEF at approximately 44 kDa. The expected band size for PSE/SPDEF is at 38 kDa.

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Figure 2. IF analysis of PSE/SPDEF using anti-PSE/SPDEF antibody (A04625-1) and anti-Tubulin Alpha antibody (M03989-3).
PSE/SPDEF was detected in immunocytochemical section of MCF-7 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-PSE/SPDEF Antibody (A04625-1) 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 MCF-7 cells using anti-PSE/SPDEF antibody (A04625-1).
Overlay histogram showing MCF-7 cells stained with A04625-1 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-PSE/SPDEF Antibody (A04625-1, 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.

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Figure 4. Flow Cytometry analysis of PC-3 cells using anti-PSE/SPDEF antibody (A04625-1).
Overlay histogram showing PC-3 cells stained with A04625-1 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-PSE/SPDEF Antibody (A04625-1, 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 SPDEF (Source: Uniprot.org, NCBI)

Alternative Names

bA375E1.3; PDEFRP11-375E1__A.3; Prostate epithelium-specific Ets transcription factor; Prostate-derived Ets factor; Prostate-specific Ets; PSE; SAM pointed domain containing ets transcription factor; SAM pointed domain-containing Ets transcription factor SPDEF PDEF, bA375E1.3 SAM pointed domain containing ETS transcription factor SAM pointed domain-containing Ets transcription factor|prostate epithelium-specific Ets transcription factor|prostate-derived Ets factor|prostate-specific Ets

For more info on SPDEF, check out the SPDEF Infographic

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In this infographic, you will see the following information for SPDEF: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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