Anti-ERp57 Antibody Picoband™ (monoclonal, 7E5)

Product Name

Anti-ERp57 Antibody Picoband™ (monoclonal, 7E5)

View all ERp57/PDIA3 Antibodies

SKU/Catalog Number

M01464-4

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-ERp57 Antibody Picoband™ (monoclonal, 7E5) catalog # M01464-4. Tested in Flow Cytometry, IHC, 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-ERp57 Antibody Picoband™ (monoclonal, 7E5) (Boster Biological Technology, Pleasanton CA, USA, Catalog # M01464-4)

Host

Mouse

Contents

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

Clonality

Monoclonal

Clone Number

7E5

Isotype

Mouse IgG1

Immunogen

A synthetic peptide corresponding to a sequence at the C-terminus of human ERp57, 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

M01464-4 is reactive to PDIA3 in Human

Applications

M01464-4 is guaranteed for Flow Cytometry, IHC, WB Boster Guarantee

Observed Molecular Weight

57 kDa

Calculated molecular weight

56.782kDa

Background of ERp57/PDIA3

PDIA3 (Protein disulfide isomerase family A, member 3), also called GRP58, Erp57 or ER60, is an isomerase enzyme. It is mapped on 15q15.3. PDIA3 is also part of the major histocompatibility complex (MHC) class I peptide-loading complex, which is essential for formation of the final antigen conformation and export from the endoplasmic reticulum to the cell surface. This gene encodes a protein of the endoplasmic reticulum that interacts with lectin chaperones calreticulin and calnexin to modulate folding of newly synthesized glycoproteins. The protein was once thought to be a phospholipase; however, it has been demonstrated that the protein actually has protein disulfide isomerase activity. It is thought that complexes of lectins and this protein mediate protein folding by promoting formation of disulfide bonds in their glycoprotein substrates.

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
Immunohistochemistry(Paraffin-embedded Section), 2-5 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 ERp57 using anti-ERp57 antibody (M01464-4).
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 placenta tissue lysates,
Lane 2: human A549 whole cell lysates,
Lane 3: human SW620 whole cell lysates,
Lane 4: human HEK293 whole cell lysates,
Lane 5: human K562 whole cell lysates,
Lane 6: human Raji whoe 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-ERp57 antigen affinity purified monoclonal antibody (Catalog # M01464-4) 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 1:5000 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 ERp57 at approximately 57 kDa. The expected band size for ERp57 is at 57 kDa.

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Figure 2. IHC analysis of ERp57 using anti-ERp57 antibody (M01464-4).
ERp57 was detected in a paraffin-embedded section of human rectal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml mouse anti-ERp57 Antibody (M01464-4) overnight at 4°C. Peroxidase Conjugated Goat Anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Mouse IgG Super Vision Assay Kit (Catalog # SV0001) with DAB as the chromogen.

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Figure 3. Flow Cytometry analysis of U87 cells using anti-ERp57 antibody (M01464-4).
Overlay histogram showing U87 cells stained with M01464-4 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-ERp57 Antibody (M01464-4, 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 PDIA3 (Source: Uniprot.org, NCBI)

Alternative Names

Disulfide isomerase ER-60; EC 5.3.4.1; endoplasmic reticulum P58; Endoplasmic reticulum resident protein 57; Endoplasmic reticulum resident protein 60; ER protein 60; ER60; ERp57; ERp5758 kDa glucose-regulated protein; ERp60; ERp6058 kDa microsomal protein; ERp61; glucose regulated protein, 58kDa; GRP57; GRP58; GRP58ER protein 57; HsT17083; P58; PDIA3; phospholipase C-alpha; PI-PLC; protein disulfide isomerase family A, member 3; protein disulfide isomerase-associated 3; protein disulfide-isomerase A3 PDIA3 ER60, ERp57, ERp60, ERp61, GRP57, GRP58, HEL-S-269, HEL-S-93n, HsT17083, P58, PI-PLC protein disulfide isomerase family A member 3 protein disulfide-isomerase A3|58 kDa glucose-regulated protein|58 kDa microsomal protein|ER protein 57|ER protein 60|disulfide isomerase ER-60|endoplasmic reticulum P58|endoplasmic reticulum resident protein 57|endoplasmic reticulum resident protein 60|epididymis secretory protein Li 269|epididymis secretory sperm binding protein Li 93n|glucose regulated protein, 58kDa|phospholipase C-alpha|protein disulfide isomerase-associated 3

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