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Product Info Summary
| SKU: | PB9512 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | Flow Cytometry, IF, ICC, WB |
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Product info
Product Name
Anti-STAT5b Antibody Picoband®
SKU/Catalog Number
PB9512
PB0541 is an alternative SKU for this antibody, used in previous lots.
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-STAT5b Antibody Picoband® catalog # PB9512. Tested in Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human, Mouse, Rat. 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
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-STAT5b Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9512)
Host
Rabbit
Contents
Each vial contains antibody formulated with stabilizing components, 0.9 mg NaCl, 0.2 mg Na2HPO4, and 0.05 mg NaN3.
*This antibody is supplied in a stabilized formulation.
Compatibility with conjugation reactions depends on the chemistry of the conjugation method used.
For conjugation methods that are not compatible with the stabilizing components present in this formulation, a carrier-free antibody format is required.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
A synthetic peptide corresponding to a sequence at the C-terminus of human STAT5b, different from the related mouse and rat sequences by two amino acids.
Cross-reactivity
No cross-reactivity with other proteins
Reactive Species
PB9512 is reactive to STAT5B in Human, Mouse, Rat
Observed Molecular Weight
90 kDa
Calculated molecular weight
89.9 kDa
Background of STAT5B
STAT5B (Signal transducer and activator of transcription 5B) is a protein that in humans is encoded by the STAT5B gene. STAT5B orthologs have been identified in most placentals for which complete genome data are available. By FISH, Lin et al. (1996) mapped STAT5B to 17q11.2. The protein encoded by this gene is a member of the STAT family of transcription factors. In response to cytokines and growth factors, STAT family members are phosphorylated by the receptor associated kinases, and then form homo- or heterodimers that translocate to the cell nucleus where they act as transcription activators. This protein mediates the signal transduction triggered by various cell ligands, such as IL2, IL4, CSF1, and different growth hormones. It has been shown to be involved in diverse biological processes, such as TCR signaling, apoptosis, adult mammary gland development, and sexual dimorphism of liver gene expression. This gene was found to fuse to retinoic acid receptor-alpha (RARA) gene in a small subset of acute promyelocytic leukemias (APML). The dysregulation of the signaling pathways mediated by this protein may be the cause of the APML.
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
PB9512 is guaranteed for Flow Cytometry, IF, ICC, 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, 0.1-0.5μg/ml, Human, Mouse, Rat
Immunocytochemistry/Immunofluorescence, 2μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
Positive Control
WB: K562 Whole Cell, HEK293 Whole Cell, PANC-1 Whole Cell, SW620 Whole Cell, SGC-7901 Whole Cell, Rat Lung TissueRat Liver TissueMouse Spleen TissueMouse Spleen TissueMouse Lung Tissue
ICC/IF: U20S cell
FCM: Jurkat cell, A431 cell
Validation Images & Assay Conditions
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Western blot analysis of STAT5b using anti-STAT5b antibody (PB9512).
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 50ug of sample under reducing conditions.
Lane 1: K562 Whole Cell Lysate,
Lane 2: HEK293 Whole Cell Lysate,
Lane 3: PANC-1 Whole Cell Lysate,
Lane 4: SW620 Whole Cell Lysate,
Lane 5: SGC-7901 Whole Cell Lysate,
Lane 6: Rat Lung Tissue Lysate.
Lane 7: Rat Liver Tissue Lysate.
Lane 8: Mouse Spleen Tissue Lysate.
Lane 9: Mouse Spleen Tissue Lysate.
Lane 10: Mouse Lung Tissue Lysate.
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 rabbit anti-STAT5b antigen affinity purified polyclonal antibody (Catalog # PB9512) 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 1:10000 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 STAT5b at approximately 90KD. The expected band size for STAT5b is at 90KD.
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Flow Cytometry analysis of Jurkat cells using anti-STAT5b antibody (PB9512).
Overlay histogram showing Jurkat cells stained with PB9512 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-STAT5b Antibody (PB9512, 1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
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Flow Cytometry analysis of A431 cells using anti-STAT5b antibody (PB9512).
Overlay histogram showing A431 cells stained with PB9512 (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 rabbit anti-STAT5b Antibody (PB9512, 1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
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IF analysis of STAT5b using anti-STAT5b antibody (PB9512).
STAT5b was detected in immunocytochemical section of U20S cells. 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 2μg/mL rabbit anti-STAT5b Antibody (PB9512) overnight at 4°C. DyLight®594 Conjugated Goat Anti-Rabbit IgG (BA1142) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Specific Publications For Anti-STAT5b Antibody Picoband® (PB9512)
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1 Customer Q&As for Anti-STAT5b Antibody Picoband®
Question
We are currently using anti-STAT5b antibody PB9512 for human tissue, and we are content with the Flow Cytometry results. The species of reactivity given in the datasheet says human, rat. Is it likely that the antibody can work on primate tissues as well?
Verified Customer
Verified customer
Asked: 2019-01-16
Answer
The anti-STAT5b antibody (PB9512) has not been tested for cross reactivity specifically with primate tissues, but there is a good chance of cross reactivity. We have an innovator award program that if you test this antibody and show it works in primate you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2019-01-16
