STAT5A Antibody (251619) [Unconjugated]

Western blot shows lysates of K562 human chronic myelogenous leukemia cell line and M1 mouse myeloid leukemia cell line. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human/Mouse STAT5a Monoclonal Antibody (Catalog # MAB2174) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for STAT5a at approximately 91 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

STAT5a was detected in immersion fixed K562 human chronic myelogenous leukemia cell line using Mouse Anti-Human/Mouse STAT5a Monoclonal Antibody (Catalog # MAB2174) at 25 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Effects of inhibition of IGF-IR on IGF-IR, BCR-ABL and downstream target proteins in CML cell lines. (A) PPP induces concentration-dependent decrease in IGF-IR tyrosine kinase activity in K562 and KBM-5 cell lines. In contrast, PPP fails to cause similar effect in BCR-ABL tyrosine kinase activity. The results represent the means ± S.D. of three consistent experiments. *: P < 0.01 and †: P < 0.001 compared with control untreated cells. (B) Western blotting and co-immunoprecipitation studies confirm that PPP decreases the tyrosine phosphorylation of IGF-IR in a concentration-dependent fashion (results shown are representative and were obtained from the KBM-5 cell line). The basal levels of IGF-IR did not change after treatment with PPP. The phosphorylation level of BCR-ABL remains unchanged after treatment with PPP. The decrease in pIGF-IR is associated with down-regulation of pAkt and pSTAT5, two oncogenic proteins in CML. Changes are not seen in Akt and STAT5. Also, PPP induces changes consistent with apoptotic cell death including down-regulation of Bcl-2, Bcl-XL and caspase-3. Moreover, treatment with PPP induces up-regulation of cyclin B1 and down-regulation of cyclin E and pCdc2, whereas the levels of Cdc2 and p16 remain unchanged. Overall, the changes in the cell cycle regulatory proteins are consistent with G2/M-phase cell cycle arrest. beta -Actin shows equal loading of the proteins. (C) IGF-IR siRNA decreases IGF-IR levels in the KBM-5 cell line and this decrease is associated with down-regulation of pAkt and pSTAT5. Basal levels of these two proteins remain unchanged. IGF-IR siRNA did not induce alterations in BCR-ABL or pBCR-ABL protein. beta -Actin confirms equal loading of the proteins. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/19508387), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu, Mu
• Applications: WB, ICC/IF
• Clone: 251619
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: LYOPH

STAT5A Antibody (251619) [Unconjugated] Summary

• Immunogen: Human STAT5a synthetic peptide
  SLDSRLSPPAGLFTSARGSLS
  Accession # NP_003143
• Specificity: Detects human and mouse STAT5a. Does not cross-react with STAT5b.
• Source: N/A
• Isotype: IgG3
• Clonality: Monoclonal
• Host: Mouse
• Gene: STAT5A
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 8-25 ug/mL
  • Western Blot 2 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for STAT5A Antibody (251619) [Unconjugated]

• MGF
• signal transducer and activator of transcription 5A
• STAT5
• STAT5a

Background

STAT family proteins mediate cytokine signaling by acting as signal transducers in the cytoplasm and transcription activators in the nucleus. STAT5a and STAT5b are encoded by separate genes and share 93% amino acid identity.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for STAT5A Antibody (MAB2174)(7)

Publications using MAB2174

• Meah, A;Vedarethinam, V;Bronstein, R;Gujarati, N;Jain, T;Mallipattu, SK;Li, Y;Wang, J; Single-Cell Spatial MIST for Versatile, Scalable Detection of Protein Markers Biosensors 2023-08-27 [PMID: 37754086] (IHC, Mouse)
• Iván Parra-Izquierdo, Alexander R. Melrose, Jiaqing Pang, Hari Hara Sudhan Lakshmanan, Stéphanie E. Reitsma, Sai Hitesh Vavilapalli et al. Janus kinase inhibitors ruxolitinib and baricitinib impair glycoprotein-VI mediated platelet function Platelets 4/3/2022 [PMID: 34097573]
• Iván Parra-Izquierdo, Alexander R. Melrose, Jiaqing Pang, Hari Hara Sudhan Lakshmanan, Stéphanie E. Reitsma, Sai Hitesh Vavilapalli et al. Janus kinase inhibitors ruxolitinib and baricitinib impair glycoprotein-VI mediated platelet function Platelets 2022-04-03 [PMID: 34097573] (Western Blot, Immunocytochemistry, Human)
• H Zhu, RH Blum, D Bernareggi, EH Ask, Z Wu, HJ Hoel, Z Meng, C Wu, KL Guan, KJ Malmberg, DS Kaufman Metabolic Reprograming via Deletion of CISH in Human iPSC-Derived NK Cells Promotes In�Vivo Persistence and Enhances Anti-tumor Activity Cell Stem Cell, 2020-06-11;0(0):. 2020-06-11 [PMID: 32531207] (Western Blot, Human)
• Sivareddy Kotla, Nikhlesh K. Singh, James G. Traylor, A. Wayne Orr, Gadiparthi N. Rao ROS-dependent Syk and Pyk2-mediated STAT1 activation is required for 15(S)-hydroxyeicosatetraenoic acid-induced CD36 expression and foam cell formation Free Radical Biology and Medicine 11/1/2014 [PMID: 25152235]
• Ping Shi, Joya Chandra, Xiaoping Sun, Mate Gergely, Jorge E. Cortes, Guillermo Garcia-Manero et al. Inhibition of IGF-IR tyrosine kinase induces apoptosis and cell cycle arrest in imatinib-resistant chronic myeloid leukaemia cells Journal of Cellular and Molecular Medicine 6/11/2009 [PMID: 19508387]
• Bao YC, Nomura Y, Moon Y, Tonozuka Y, Minoshima Y, Hatori T, Tsuchiya A, Kiyono M, Nosaka T, Nakajima H Rac1 and a GTPase-activating protein, MgcRacGAP, are required for nuclear translocation of STAT transcription factors. J. Cell Biol., 2006-12-18;175(6):937-46. 2006-12-18 [PMID: 17178910] (ICC, Western Blot, Human, Mouse)

Review for STAT5A Antibody (MAB2174) (1)

Average Rating: 3

(Based on 1 review)

We have 1 review tested in 1 species: Human.

reviewed by: Verified Customer Simple Western Human 11/28/2016

Summary

• Application: Simple Western
• Sample Tested: k562 cells
• Species: Human

Comments

• Comments: Theoretical pI = 5.98; no peak close to that value
  
  Cells used = K562 (human) lysed in Protein Simple bicine/CHAPS buffer (040-764) loaded at 100 ng/µL; Gradient = 5 – 8 nested, ladder# 3
  
  Primary antibody at 1:50
  
  Secondary antibody = Protein Simple biotinylated anti-mouse (041-127) with Protein Simple streptavidin-HRP (041-106) both at 1:100
  
  Predicted m.w. = 90.6 kDa; well defined peak at 95 kDa, good agreement between replicates
  
  Cells used = K562 (human) lysed in Protein Simple bicine/CHAPS buffer (040-764) loaded at 500 ng/µL
  
  Primary antibody at 1:50

Bioinformatics

• Gene Symbol: STAT5A
• Uniprot:
  • NP_003143()