Survivin Overexpression Lysate

Western Blot: Survivin Overexpression Lysate [NBL1-07987] - Western blot validation with an anti-DDK antibody. Left: Empty vector transfected control cell lysate (HEK293 cell lysate); Right: Survivin Overexpression Lysate [NBL1-07987].

• Reactivity: Hu
• Applications: WB

Survivin Overexpression Lysate Summary

• Description:

  Survivin Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC205935
  
  Accession#: NM_001168
  
  Protein Tag: C-MYC/DDK
  
  You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.
• Gene: BIRC5

Applications/Dilutions

• Dilutions:
  • Western Blot
• Application Notes: This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct.
  
  Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.
• Theoretical MW: 16.2 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• Storage: Store at -80C. Avoid freeze-thaw cycles.
• Buffer: RIPA buffer

Lysate Details for Array

• Type: Overexpression

Notes

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

Alternate Names for Survivin Overexpression Lysate

• API4
• apoptosis inhibitor 4
• Apoptosis inhibitor survivin
• baculoviral IAP repeat-containing 5
• Baculoviral IAP Repeat-Containing Protein 5
• BIRC5
• EPR-1
• IAP4
• survivin variant 3 alpha
• Survivin

Background

Survivin (BIRC5 or IAP4) is the smallest member of the inhibitor of apoptosis (IAP) proteins, containing only a single baculovirus IAP repeat (BIR) domain and lacking the C-terminal RING domain. Survivin has a theoretical molecular weight of 16.5 kDa (wild-type protein) and exists as ten splice variant forms including the most predominant isoforms, survivin-2B and surviving-DeltaEx3 (1). It also undergoes phosphorylation by PKA, Plk1, Cdk1, CKII and Aurora B at multiple sites (e.g. Serine 20 and Threonine 34, 53 and 117).

Besides being highly abundant in fetal development and expressed in proliferating adult cells such as activated T lymphocytes, erythroblasts, and self-renewing stem cells, survivin is generally absent in adult tissues. However, it is elevated in common cancers such as lung, colon, pancreas, breast and prostate where it drives proliferation, metastasis, poor prognosis, and decreased patient survival (2).

Survivin has been shown to be involved in multiple cellular processes including cell cycle progression, mitotic spindle assembly, kinetochore attachment, angiogenesis, migration, and its anti-apoptotic activity has been linked to both its monomeric and homodimeric forms. Survivin impacts the function of other IAP members, c-IAP1 and c-IAP-2, or modulates the inhibitory activity of XIAP against caspases by forming a stable complex with XIAP and HBXIP. During the intrinsic apoptotic pathway, survivin may prevent the release of mitochondrial APAF1 into the cytoplasm or hinder the association of SMAC with other IAPS, which results in prolonged cell survival (3).

References

1. Sah NK, Seniya C. (2015) Survivin splice variants and their diagnostic significance. Tumour Biol. 36(9):6623-31. PMID: 26245993

2. Lladser A, Sanhueza C, Kiessling R, Quest AF. (2011) Is survivin the potential Achilles' heel of cancer? Adv Cancer Res. 111:1-37. PMID: 21704829

3. Wheatley SP, Altieri DC. (2019) Survivin at a glance. J Cell Sci. 132(7). PMID: 30948431

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

FAQs for Survivin Lysate (NBL1-07987). (Showing 1 - 3 of 3 FAQ).

1. Can I use this antibody with species other than those listed?
   • The species we have listed are validated and therefore have a 100% guarantee to work with this antibody. We cannot guarantee that this will work with other species.
2. Does this lysate have a tag?
   • Yes, the overexpressed protein contains a C-terminal DDK tag.
3. Can this be used as a control lysate?
   • Yes, this lysate is intended for use as a positive control in Western Blot.

Bioinformatics

• Gene Symbol: BIRC5