Product Datasheet. HIF-1 alpha Antibody NB Unit Size: 0.1 ml

Transcription

1 Product Datasheet HIF-1 alpha Antibody NB Unit Size: 0.1 ml Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles. Reviews: 13 Publications: 139 Protocols, Publications, Related Products, Reviews, Research Tools and Images at: Updated 10/28/2018 v.20.1 Earn rewards for product reviews and publications. Submit a publication at Submit a review at

2 NB HIF-1 alpha Antibody Product Information Unit Size Concentration Storage Clonality Preservative Isotype Purity Buffer Target Molecular Weight 0.1 ml 1.0 mg/ml Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles. Polyclonal 0.05% Sodium Azide IgG Immunogen affinity purified PBS 93 kda Page 1 of 7 v.20.1 Updated 10/28/2018 Product Description Host Rabbit Gene ID 3091 Gene Symbol Species Reactivity Notes Immunogen HIF1A Human, Mouse, Rat, Canine, Guinea Pig, Primate, Xenopus, Zebrafish Canine reactivity was reported in scientific literature (PMID: ). Mouse reactivity results are mixed. Cross-reactivity with Zebrafish was reported via a customer review. A fusion protein made to an internal sequence of human HIF-1 alpha (containing amino acids ) [UniProt Q16665]. Product Application Details Applications Recommended Dilutions Application Notes Western Blot, Simple Western, Gel Super Shift Assays, Immunocytochemistry/Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Immunoprecipitation, Knockout Validated Western Blot 1:500-1:1000, Simple Western 1:100, Immunohistochemistry 1:100-1:500, Immunocytochemistry/Immunofluorescence 1:100-1:500, Immunoprecipitation 1:1000, Immunohistochemistry-Paraffin 1:100-1:500, Immunohistochemistry-Frozen 1:100-1:500, Gel Super Shift Assays 1:1-1:100, Knockout Validated ChIP usage was reported in scientific literature (PMID: ). ICC/IF usage was reported in scientific literature (PMID: , ). In WB a band is observed approx. 120 kda. IHC applications have been mixed on Mouse. For ChIP, nuclear extracts are recommended. Prior to immunostaining paraffin tissues, antigen retrieval with sodium citrate buffer (ph 6.0) is recommended. In Simple Western only ul of the recommended dilution is used per data point. Knock Out Validation was reported in scientific literature (PMID: ).

3 Images Western Blot: HIF-1 alpha Antibody [NB ] - Analysis of HIF-1 alpha in overexpression and endogenous HIF1 alpha & HIF2 alpha using anti-hif-1 alpha antibody. The data showed that HIF-1 alpha antibody did not react to HIF-2 alpha overexpression. Image from verified customer review. Page 2 of 7 v.20.1 Updated 10/28/2018 Western Blot: HIF-1 alpha Antibody [NB ] - Analysis of NB , anti-hif-1 alpha, on normoxic and hypoxic nuclear rat cell lysates. Immunoprecipitation: HIF-1 alpha Antibody [NB ] - PC-3 Cell Lysates run with Cobalt Chloride treatments. Immunoprecipitation was performed with protein A/G Agarose beads. This data was provided courtesy of Kelie Reece, Figg lab, NCI. Immunocytochemistry/Immunofluorescence: HIF-1 alpha Antibody [NB ] - Analysis of HIF-1 alpha in ARPE-19 cells using anti-hif-1 alpha antibody. Image from verified customer review.

4 Immunohistochemistry-Paraffin: HIF-1 alpha Antibody [NB ] - Staining in the canine CL on days 10 to 70 after ovulation. PC = positive control (human placenta). NC = negative control. Image from verified customer review. Page 3 of 7 v.20.1 Updated 10/28/2018 Simple Western: HIF-1 alpha Antibody [NB ] - Image shows a specific band for HIF-1 alpha in 0.2 mg/ml of Hypoxic HeLa lysate. This experiment was performed under reducing conditions using the kda separation system. Immunohistochemistry-Paraffin: HIF-1 alpha Antibody [NB ] - Analysis of FFPE tissue section of human endometrium carcinoma AN3CA cell line based xenograft using rabbit polyclonal HIF1 alpha antibody at 1:300 dilution. The signal was developed using HRP-labelled secondary antibody and DAB reagent, and the section was further counterstained using hematoxylin. The tested section depicted mainly a diffused cytoplasmic staining but there were some cells which showed nuclear signal also (representing hypoxic cells). Immunohistochemistry-Paraffin: HIF-1 alpha Antibody [NB ] - Analysis of HIF-1 alpha on Zebrafish retina tissue using HIF-1 alpha antibody. Mouse tissue was used as a control. Image from verified customer review.

5 Western Blot: HIF-1 alpha Antibody [NB ] - 20 ug whole cell lysates (human ovarian cancer cell lines). Image from verified customer review. Page 4 of 7 v.20.1 Updated 10/28/2018 Western Blot: HIF-1 alpha Antibody [NB ] - Analysis of HIF-1 alpha in human Retinal and Choroidal primary endothelia cells using anti-hif-1 alpha antibody. Image from verified customer review. Immunohistochemistry: HIF-1 alpha Antibody [NB ] - Staining of human kidney, renal tubular epithelium in Cortex.

6 Publications Zhang X, Chen L. Effects of CoCl2-simulated hypoxia on the expression levels of matrix metalloproteinases in renal adenocarcinoma cells and renal tubular epithelial cells Exp Ther Med Aug :00AM [PMID: ] (WB, Human) Ronellenfitsch MW, Zeiner PS, Mittelbronn M et al. Akt and mtorc1 signaling as predictive biomarkers for the EGFR antibody nimotuzumab in glioblastoma Acta Neuropathol Commun Aug :00AM [PMID: ] (IHC, Human) Mazumdar Jolly, O'brien W Timothy, Johnson Randall S et al. O2 regulates stem cells through Wnt/B-catenin signalling. Nature Cell Biology 2010 [PMID: ] (ChIP, Mouse) McClendon J, Jansing NL, Redente EF et al. Hypoxia-Inducible Factor 1a Signaling Promotes Repair of the Alveolar Epithelium after Acute Lung Injury. Am. J. Pathol. Aug :00AM [PMID: ] (WB, Mouse) Zheng J, Zhang M, Weng H. Induction of the mitochondrial NDUFA4L2 protein by HIF-1a regulates heart regeneration by promoting the survival of cardiac stem cell. Biochem. Biophys. Res. Commun. Jun :00AM [PMID: ] (ChIP, Human) Chhipa RR, Fan Q, Anderson J et al. AMP kinase promotes glioblastoma bioenergetics and tumour growth. Nat. Cell Biol. Jul :00AM [PMID: ] (ChIP, Human) Peruzzotti-Jametti L, Bernstock JD, Vicario N et al. Macrophage-Derived Extracellular Succinate Licenses Neural Stem Cells to Suppress Chronic Neuroinflammation. Cell Stem Cell. Mar :00AM [PMID: ] (WB) Trollmann R1, Strasser K, Keller S et al. Placental HIFs as markers of cerebral hypoxic distress in fetal mice. Am J Physiol Regul Integr Comp Physiol Dec [PMID: ] (WB, Mouse) Eriksson JA, Wanka C, Burger MC et al. Suppression of oxidative phosphorylation confers resistance against bevacizumab in experimental glioma J. Neurochem Nov 27 [PMID: ] (Human) Li D, Du Y, Yuan X et al. Hepatic hypoxia-inducible factors inhibit PPARalpha expression to exacerbate acetaminophen induced oxidative stress and hepatotoxicity. Free Radic. Biol. Med Jun 03 [PMID: ] Zhang X, Liu L, Liu J et al. Endothelial cells co-cultured with renal carcinoma cells significantly reduce RECK expression under chemical hypoxia. Cell Biol. Int May 31 [PMID: ] (Human) Jang JE, Ko MS, Yun JY et al. Nitric Oxide Produced by Macrophages Inhibits Adipocyte Differentiation and Promotes Profibrogenic Responses in Preadipocytes to Induce Adipose Tissue Fibrosis. Diabetes 2016 Sep [PMID: ] (WB, Mouse) More publications at Page 5 of 7 v.20.1 Updated 10/28/2018

7 Page 6 of 7 v.20.1 Updated 10/28/2018 Procedures Western blot Protocol specific for HIF-1 alpha antibody (NB ) Western Blot Protocol 1. Perform SDS-PAGE on hypoxic samples to be analyzed, loading 30 ug of nuclear protein per lane. 2. Transfer proteins to membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus. 3. Stain according to standard Ponceau S procedure (or similar product) to assess transfer success, and mark molecular weight standards where appropriate. 4. Rinse the blot. 5. Block the membrane using standard blocking buffer for at least 1 hour. 6. Wash the membrane in wash buffer three times for 10 minutes each. 7. Dilute primary antibody in blocking buffer and incubate 1 hour at room temperature. 8. Wash the membrane in wash buffer three times for 10 minutes each. 9. Apply the diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature. 10. Wash the blot in wash buffer three times for 10 minutes each (this step can be repeated as required to reduce background). 11. Apply the detection reagent of choice in accordance with the manufacturers instructions. Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of %. Immunohistochemistry-Frozen Protocol for HIF-1 alpha Antibody (NB ) Immunohistochemistry-Paraffin Embedded Sections Antigen Unmasking: Bring slides to a boil in 10 mm sodium citrate buffer (ph 6.0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes. Staining: 1. Wash sections in deionized water three times for 5 minutes each. 2. Wash sections in wash buffer for 5 minutes. 3. Block each section with ul blocking solution for 1 hour at room temperature. 4. Remove blocking solution and add ul diluted primary antibody. Incubate overnight at 4 C. 5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each. 6. Add ul biotinylated diluted secondary antibody. Incubate 30 minutes at room temperature. 7. Remove secondary antibody solution and wash sections three times with wash buffer for 5 minutes each. 8. Add ul Streptavidin-HRP reagent to each section and incubate for 30 minutes at room temperature. 9. Wash sections three times in wash buffer for 5 minutes each. 10. Add ul DAB substrate to each section and monitor staining closely. 11. As soon as the sections develop, immerse slides in deionized water. 12. Counterstain sections in hematoxylin. 13. Wash sections in deionized water two times for 5 minutes each. 14. Dehydrate sections. 15. Mount coverslips.

8 Novus Biologicals USA E. Briarwood Avenue Centennial, CO USA Phone: Toll Free: Fax: Novus Biologicals Canada 461 North Service Road West, Unit B37 Oakville, ON L6M 2V5 Canada Phone: Toll Free: Fax: Novus Biologicals Europe 19 Barton Lane Abingdon Science Park Abingdon, OX14 3NB, United Kingdom Phone: (44) (0) Free Phone: Fax: (44) (0) General Contact Information Technical Support: Orders: General: Products Related to NB NBP HAF008 NB7156 NBP HeLa Hypoxic / Normoxic Cell Lysate Goat anti-rabbit IgG Secondary Antibody [HRP (Horseradish Peroxidase)] Goat anti-rabbit IgG (H+L) Secondary Antibody Rabbit IgG Isotype Control 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. For more information on our 100% guarantee, please visit Earn gift cards/discounts by submitting a review: Earn gift cards/discounts by submitting a publication using this product: