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Product Info Summary
| SKU: | M01464-4 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human |
| Host: | Mouse |
| Application: | Flow Cytometry, IHC, WB |
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Product info
Product Name
Anti-ERp57 Antibody Picoband® (monoclonal, 7E5)
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. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
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.
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
M01464-4 is reactive to PDIA3 in Human
Observed Molecular Weight
57 kDa
Calculated molecular weight
56.8 kDa
Background of 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.
Application & Images
Applications
M01464-4 is guaranteed for Flow Cytometry, IHC, WB Boster Guarantee
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 (Fixed), 1-3 μg/1x106 cells, Human
Positive Control
WB: human placenta tissue, human A549 whole cell, human SW620 whole cell, human HEK293 whole cell, human K562 whole cell, human Raji whoe cell
IHC: human rectal cancer tissue
FCM: U87 cell
Validation Images & Assay Conditions
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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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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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Flow Cytometry analysis of U87 cells using anti-ERp57 antibody (M01464-4).
Overlay histogram showing U87 cells stained with M01464-4 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. 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 without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Specific Publications For Anti-ERp57 Antibody Picoband® (monoclonal, 7E5) (M01464-4)
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