Anti-STAT5b Antibody Picoband™

Product Name

Anti-STAT5b Antibody Picoband™

View all STAT5B Antibodies

SKU/Catalog Number

PB9512

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.

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 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.

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.

*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

PB9512 is reactive to STAT5B in Human, Mouse, Rat

Applications

PB9512 is guaranteed for Flow Cytometry, IF, ICC, WB Boster Guarantee

Observed Molecular Weight

90 kDa

Calculated molecular weight

89.866kDa

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.

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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, 0.1-0.5μg/ml, Human, Mouse, Rat
Immunocytochemistry/Immunofluorescence, 2μg/ml, Human
Flow Cytometry, 1-3μg/1x10⁶ cells, Human

Validation Images & Assay Conditions

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Figure 1. 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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Figure 2. 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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Figure 3. Flow Cytometry analysis of A431 cells using anti-STAT5b antibody (PB9512).
Overlay histogram showing A431 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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Figure 4. 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.

Gene/Protein Information For STAT5B (Source: Uniprot.org, NCBI)

Alternative Names

signal transducer and activator of transcription 5B; STAT5; STAT5b; transcription factor STAT5B STAT5B GHISID2, STAT5 signal transducer and activator of transcription 5B signal transducer and activator of transcription 5B|transcription factor STAT5B

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In this infographic, you will see the following information for STAT5B: 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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