bcl-x Antibody [Unconjugated]

Western blot shows lysates of A431 human epithelial carcinoma cell line (5 x 104cells, lane 1 and 2.5 x 104cells, lane 2) and NIH-3T3 mouse embryonic fibroblast cell line. PVDF membrane was probed with 0.3 µg/mL of Rabbit Anti-Human Bcl-x Antigen Affinity-purified Polyclonal Antibody (Catalog # AF800) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Bcl-x at approximately 26 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.

Bcl-x was immunoprecipitated from lysates (7 x 105cells) of A431 human epithelial carcinoma cell line following incubation with 3 µg Rabbit Anti-Human Bcl‑x Antigen Affinity-purified Polyclonal Antibody (Catalog # AF800) for 30‑60 minutes at 4 °C. Bcl-x-antibody complexes were absorbed using Protein A Immunoprecipitin (Life Technologies). Immuno-precipitated Bcl-x was detected by Western blot using 0.3 µg/mL Rabbit Anti-Human Bcl‑x Antigen Affinity-purified Polyclonal Antibody (Catalog # AF800). View our recommended buffer recipes for immunoprecipitation.

Simple Western lane view shows lysates of A431 human epithelial carcinoma cell line and NIH‑3T3 mouse embryonic fibroblast cell line, loaded at 0.2 mg/mL. A specific band was detected for Bcl‑x at approximately 34 kDa (as indicated) using 3 µg/mL of Rabbit Anti-Human/Mouse Bcl‑x Antigen Affinity-purified Polyclonal Antibody (Catalog # AF800). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Necroptosis induction in parental CNE-2 and photon-resistant CNE-2-RR cells following X-rays or carbon-ion irradiation. (A) Endogenous changes of CNE-2-RR in the levels of caspase-8, PARP and other regulators of necroptosis compared with CNE-2 cells. Protein levels (mean±SD) in CNE-2-RR relative to the CNE-2 cells are presented. **P≤0.01. (B) Dose dependent phosphorylation of MLKL in X-ray irradiated cells at 48 hours. (C) Dose dependent phosphorylation of MLKL, PARP and cleaved-PARP in carbon ion irradiated cells at 48 hours. (D) Levels of phosphorylated forms MLKL and Bcl-x at 48 hours after X-ray (12 Gy) and carbon-ion (4 Gy) irradiation in HepG2 and Hep2 cells. (E) Levels of phosphorylated forms MLKL and Bcl-x at 48 hours after X-ray (4 Gy and 10 Gy) and carbon-ion (2 Gy and 4 Gy) irradiation in CNE-2 and CNE-2-RR cells. (F) Fold change in p-MLKL levels (mean ± SEM) in irradiated cells (Carbon ion: 2Gy and 4 Gy; X-ray: 4Gy and 10Gy) relative to the un-irradiated cells at 48 h. **P≤0.01, ****P≤0.0001. ns, no significance. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/33531997), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu, Mu
• Applications: WB, Simple Western, IP
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: LYOPH

bcl-x Antibody [Unconjugated] Summary

• Immunogen: KLH-coupled Bcl-x synthetic peptide
  CSAINGNPSWHLADSPAVNGA
• Specificity: Detects human and mouse Bcl‑x in Western blots.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: BCL2L1
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunoprecipitation 5 ug/10^6 cells
  • Simple Western 3 ug/mL
  • Western Blot 0.3 ug/mL
• Application Notes: In Simple Western only 10-15 uL of the recommended dilution is used per data point.

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 0.2 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 bcl-x Antibody [Unconjugated]

• Apoptosis regulator Bcl-X;BCL2L;BCL2L1;Bcl-2-like protein 1;BCLX;Bcl-X;BCL-XL/S;PPP1R52
• BCL2L1
• Bclx
• Bcl-x

Background

The peptide corresponds to amino acids 49-68 of human and mouse, long and short forms of Bcl-x. The sequence used for immunization differs significantly from the sequences of the corresponding region of other members of the Bcl-2 family. Cysteine was added to the amino-terminal for coupling to KLH and for coupling to an affinity matrix.

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 bcl-x Antibody (AF800)(11)

Publications using AF800

• M Kimsa-Dude, A Synowiec-W, A Krawczyk, A Kosowska, M Kimsa-Furd, T Francuz The Apoptotic Effect of Caffeic or Chlorogenic Acid on the C32 Cells That Have Simultaneously Been Exposed to a Static Magnetic Field International Journal of Molecular Sciences, 2022-03-31;23(7):. 2022-03-31 [PMID: 35409218] (Western Blot, Human)
• C Bao, Y Sun, B Dwarakanat, Y Dong, Y Huang, X Wu, C Guha, L Kong, JJ Lu Carbon ion triggered immunogenic necroptosis of nasopharyngeal carcinoma cells involving necroptotic inhibitor BCL-x Journal of Cancer, 2021-01-01;12(5):1520-1530. 2021-01-01 [PMID: 33531997] (Western Blot, Human)
• Yu H, Ding J, Zhu H et al. LOXL1 confers antiapoptosis and promotes gliomagenesis through stabilizing BAG2 Cell Death Differ. 5/18/2020 [PMID: 32424143]
• N S Seng, J Megyesi, A Tarcsafalvi, P M Price Mimicking Cdk2 phosphorylation of Bcl-xL at Ser73 results in caspase activation and Bcl-xL cleavage Cell Death Discovery 5/31/2016 [PMID: 27239332]
• J Megyesi, A Tarcsafalvi, NSHL Seng, R Hodeify, P M Price Cdk2 phosphorylation of Bcl-xL after stress converts it to a pro-apoptotic protein mimicking Bax/Bak Cell Death Discovery 5/27/2016 [PMID: 27226901]
• Schmitt E, Beauchemin M, Bertrand R Nuclear colocalization and interaction between bcl-xL and cdk1(cdc2) during G2/M cell-cycle checkpoint. Oncogene, 2007-03-19;26(40):5851-65. 2007-03-19 [PMID: 17369848] (Immunoprecipitation, Western Blot, Human)
• Armstrong T, Murphy LB, Bateman AC, Conti JA, Fine DR, Johnson CD, Benyon RC, Iredale JP Type I collagen promotes the malignant phenotype of pancreatic ductal adenocarcinoma. Clin. Cancer Res., 2004-11-01;10(21):7427-37. 2004-11-01 [PMID: 15534120] (Western Blot, Human)
• Mullen P, McPhillips F, MacLeod K, Monia B, Smyth JF, Langdon SP Antisense oligonucleotide targeting of Raf-1: importance of raf-1 mRNA expression levels and raf-1-dependent signaling in determining growth response in ovarian cancer. Clin. Cancer Res., 2004-03-15;10(6):2100-8. 2004-03-15 [PMID: 15041731] (Western Blot, Human)
• Raman VS, Akondy RS, Rath S, Bal V, George A Ligation of CD27 on B cells in vivo during primary immunization enhances commitment to memory B cell responses. J. Immunol., 2003-12-01;171(11):5876-81. 2003-12-01 [PMID: 14634097] (Western Blot, Mouse)
• Sade H, Krishna S, Sarin A The anti-apoptotic effect of Notch-1 requires p56lck-dependent, Akt/PKB-mediated signaling in T cells. J. Biol. Chem., 2003-10-28;279(4):2937-44. 2003-10-28 [PMID: 14583609] (Western Blot, Human, Mouse)
• Ye Q, Press B, Kissler S, Yang XF, Lu L, Bassing CH, Sleckman BP, Jansson M, Panoutsakopoulou V, Trimble LA, Alt FW, Cantor H T cell costimulation through CD28 depends on induction of the Bcl-xgamma isoform: analysis of Bcl-xgamma-deficient mice. J. Exp. Med., 2002-07-01;196(1):87-95. 2002-07-01 [PMID: 12093873] (Western Blot, Mouse)

Review for bcl-x Antibody (AF800) (1)

Average Rating: 5

(Based on 1 review)

We have 1 review tested in 1 species: Human.

reviewed by: Verified Customer WB Human 01/09/2015

Summary

• Application: Western Blot
• Sample Tested: See PMID 22582944
• Species: Human

Bioinformatics

• Gene Symbol: BCL2L1