Key Product Details
Validated by
Knockout/Knockdown, Biological Validation
Species Reactivity
Validated:
Human, Mouse
Cited:
Human, Mouse
Applications
Validated:
Knockout Validated, Western Blot
Cited:
Western Blot
Label
Unconjugated
Antibody Source
Monoclonal Mouse IgG2A Clone # 117320
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Product Specifications
Immunogen
E. coli-derived recombinant human XIAP
Met1-Ser497
Accession # P98170
Met1-Ser497
Accession # P98170
Specificity
Detects human and mouse XIAP in direct ELISAs and Western blots. In direct ELISAs, no cross-reactivity with recombinant human (rh) cIAP-1 or rhcIAP-2 is observed.
Clonality
Monoclonal
Host
Mouse
Isotype
IgG2A
Scientific Data Images for Human/Mouse XIAP Antibody
Detection of Human/Mouse XIAP by Western Blot.
Western blot shows lysates of WS-1 human fetal skin fibroblast cell line and L-929 mouse fibrosarcoma cell line. PVDF membrane was probed with 2 µg/mL Mouse Anti-Human/Mouse XIAP Monoclonal Antibody (Catalog # MAB822) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). For additional reference, Recombinant Human XIAP Full Length (Catalog # 822-XF) and recombinant mouse XIAP (5 ng/lane) were included. A specific band for XIAP was detected at approximately 55 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 6.
Western Blot Shows Human XIAP Specificity by Using Knockout Cell Line.
Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and XIAP knockout HeLa cell line (KO). PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human/Mouse XIAP Monoclonal Antibody (Catalog # MAB822) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for XIAP at approximately 52 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. GAPDH (Catalog # MAB5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of Human XIAP by Western Blot
VP-16 had a more potent effect on LX-2 cells than normal hepatocytes.(a–c) Three cell lines were treated with 0–4 μM VP-16 for 72 h. (a) Cytotoxicity was measured using the CCK-8 assay. (b) Apoptotic cells were analyzed using flow cytometry. Annexin V-positive cells are displayed in the bar chart as a percentage of the untreated controls. (c) Apoptosis-regulating protein levels were analyzed by western blotting. beta -actin was used as a loading control. The data are presented as the mean ± SD of three independent experiments. *P < 0.05, ***P < 0.001. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/27680712), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse XIAP by Western Blot
cIAP2 and XIAP regulate autophagosome fusion, but not cIAP1.a MEFs expressing mCherry-GFP-LC3b were transfected with siRNA against either cIAP1, cIAP2 or XIAP. Cells were analysed with the microscope and the number of mCherry+, and GFP + puncta/cell were calculated and the ratio of GFP + /mCherry + puncta is indicated. Shown are the means and the error bars represent the SEM of at least three independent experiments. Westerns show efficient knockdown of cIAP1 and XIAP expression. cIAP2 siRNA efficiency was determined by real time PCR as shown in the graph below the westerns. b Wild type and cIAP2−/− XIAP−/− dermal fibroblasts expressing mCherry-GFP-LC3b were treated analysed on the microscope and mCherry+, and GFP + puncta/cell were calculated. The ratio of GFP+/mCherry + puncta is indicated. c Wild type and cIAP2−/− XIAP−/− dermal fibroblasts were left in complete media (CM) or starved for 2 h in EBSS. Cells were lysed and proteins analysed by western blot for XIAP, LC3, and Actin. cIAP2−/− was confirmed by PCR due to lack of effective antibodies for mouse cIAP2 (see supplemental Fig. 2). d Immunofluoresence showing accumulation of LC3 in starved cIAP2−/− XIAP−/− dermal fibroblasts. Wild type and cIAP2−/− XIAP−/− dermal fibroblasts were incubated in complete media or starved in EBSS for 2 h, then fixed and stained for LC3 (green channel) and LAMP2 (red channel). Nuclei are stained blue with Hoechst. Shown in upper panels are overviews. Lower panels show zoomed in regions indicated in the upper panels Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/29743550), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse XIAP by Western Blot
cIAP2 and XIAP regulate autophagosome fusion, but not cIAP1.a MEFs expressing mCherry-GFP-LC3b were transfected with siRNA against either cIAP1, cIAP2 or XIAP. Cells were analysed with the microscope and the number of mCherry+, and GFP + puncta/cell were calculated and the ratio of GFP + /mCherry + puncta is indicated. Shown are the means and the error bars represent the SEM of at least three independent experiments. Westerns show efficient knockdown of cIAP1 and XIAP expression. cIAP2 siRNA efficiency was determined by real time PCR as shown in the graph below the westerns. b Wild type and cIAP2−/− XIAP−/− dermal fibroblasts expressing mCherry-GFP-LC3b were treated analysed on the microscope and mCherry+, and GFP + puncta/cell were calculated. The ratio of GFP+/mCherry + puncta is indicated. c Wild type and cIAP2−/− XIAP−/− dermal fibroblasts were left in complete media (CM) or starved for 2 h in EBSS. Cells were lysed and proteins analysed by western blot for XIAP, LC3, and Actin. cIAP2−/− was confirmed by PCR due to lack of effective antibodies for mouse cIAP2 (see supplemental Fig. 2). d Immunofluoresence showing accumulation of LC3 in starved cIAP2−/− XIAP−/− dermal fibroblasts. Wild type and cIAP2−/− XIAP−/− dermal fibroblasts were incubated in complete media or starved in EBSS for 2 h, then fixed and stained for LC3 (green channel) and LAMP2 (red channel). Nuclei are stained blue with Hoechst. Shown in upper panels are overviews. Lower panels show zoomed in regions indicated in the upper panels Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/29743550), licensed under a CC-BY license. Not internally tested by R&D Systems.Applications for Human/Mouse XIAP Antibody
Application
Recommended Usage
Knockout Validated
XIAP
is specifically detected in HeLa human cervical epithelial carcinoma parental cell line but is not detectable in
XIAP knockout HeLa cell line.
Western Blot
2 µg/mL
Sample: WS-1 human fetal skin fibroblast cell line and L-929 mouse fibrosarcoma cell line
Sample: WS-1 human fetal skin fibroblast cell line and L-929 mouse fibrosarcoma cell line
Reviewed Applications
Read 3 reviews rated 4 using MAB822 in the following applications:
Formulation, Preparation, and Storage
Purification
Protein A or G purified from hybridoma culture supernatant
Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.
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Formulation
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.
Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
Stability & 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.
Calculators
Background: XIAP
Long Name
X-linked Inhibitor of Apoptosis
Alternate Names
BIRC4
Gene Symbol
XIAP
UniProt
Additional XIAP Products
Product Documents for Human/Mouse XIAP Antibody
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Human/Mouse XIAP Antibody
For research use only
Related Research Areas
Citations for Human/Mouse XIAP Antibody
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Customer Reviews for Human/Mouse XIAP Antibody (3)
4 out of 5
3 Customer Ratings
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Application: MicroarraySample Tested: EDTA PlasmaSpecies: HumanVerified Customer | Posted 12/06/2019Antibody was printed on custom arrays and incubated with fluorescently labeled human EDTA plasma
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Application: ELISASample Tested: PlasmaSpecies: HumanVerified Customer | Posted 11/10/2018
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Application: Western BlotSample Tested: See PMID 20074295Species: HumanVerified Customer | Posted 02/25/2015
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Cellular Response to Hypoxia Protocols
- R&D Systems Quality Control Western Blot Protocol
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
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Associated Pathways
Ubiquitination Cascade Pathway
