HIF-1 alpha Antibody [Unconjugated]

Western blot shows lysates of MCF-7 human breast cancer cell line untreated (-) or treated (+) with 150 µM CoCl2for 16 hours. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human/Mouse/Rat HIF-1a/HIF1A Antigen-affinity Purified Polyclonal Antibody, followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for HIF-1a/HIF1A at approximately 120 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Mouse primary kidney cells treated with 150 μM CoCl2 for overnight were fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. HIF‑1 alpha /DNA complexes were immunoprecipitated using 5 μg Goat Anti-Human/Mouse/Rat HIF‑1 alpha /HIF1A Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1935) or control antibody (Catalog # AB-108-C) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Goat IgG Secondary Antibody (Catalog # BAF109). Immunocomplexes were captured using 50 μL of MagCellect Streptavidin Ferrofluid (Catalog # MAG999) and DNA was purified using chelating resin solution. The epo promoter was detected by standard PCR.

HIF-1a/HIF1A was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line treated with DFO but is not detected in HIF-1a/HIF1A knockout (KO) HeLa Human Cell Line cell line using Goat Anti-Human/Mouse/Rat HIF-1a/HIF1A Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1935) at 0.3 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Simple Western lane view shows lysates of A549 human lung carcinoma cell line untreated (-) or treated (+) with Hypoxia (1% O2), loaded at 0.2 mg/mL. A specific band was detected for HIF‑1 alpha /HIF1A at approximately 115 kDa (as indicated) using 5 µg/mL of Goat Anti-Human/Mouse/Rat HIF‑1 alpha /HIF1A Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1935) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the12-230 kDa separation system.

Ten days of IH increases hippocampal HIF1a and disrupts Barnes maze performance in wild-type mice but not in HIF1a+/−. A, left, Representative digitized Western blotting images for HIF1a (103 kDa) and PCNA (40 kDa) in hippocampal nuclear protein fractions from control (n = 4) and IH10 (n = 4). Right, Quantification of HIF1a protein normalized to PCNA revealed that nuclear HIF1a was increased in IH10 when compared with control (p = 0.019). B, Total latency to exit the Barnes maze during three training sessions in control (n = 10) and in IH10 (n = 11). Each blue (control) and red (IH10) line represents an individual performance during training. Training to the exit was conducted over three sessions. Each session was separated by 24 hours. C, Left, During the probe trial, the distance traveled to initially enter the exit zone was shorter in control when compared with IH10 (p = 0.048). Right, Latency to initial entry was smaller in control as well (p = 0.034). D, Heat maps of the mean entry probability across all false exits (1–19) and the exit zone during probe trial for the control and IH10. Comparison of entry probability into the exit zone during the probe trial reveals that control has a greater probability for entering the exit zone when compared with IH10 (p = 0.004). E, Left, Representative digitized Western blotting images HIF1a and PCNA in hippocampal nuclear protein fractions from 0-HIF1a+/− (n = 4) and 10-HIF1a+/− (n = 4). Right, Quantification of HIF1a protein normalized to PCNA revealed that nuclear HIF1a is similar between 0-HIF1a+/− and 10-HIF1a+/− (p = 0.84). F, Total latency to exit the Barnes maze during three training sessions in 0-HIF1a+/− (n = 7) and in 10-HIF1a+/− (n = 8). Each gray (0-HIF1a+/−) and yellow (10-HIF1a+/−) line represents an individual performance during training. All experimental groups exhibit decreased total latency over the course of training. G, Left, In HIF1a+/−, the distance initial to initial entry into the exit zone was similar between 0-HIF1a+/− and 10-HIF1a+/− (p = 0.55). Right, Latency to initial entry into the exit zone during the probe trial were similar between 0-HIF1a+/− and 10-HIF1a+/− (p = 0.39). H, Heat maps of the mean entry probability into all zones during the probe trial for 0-HIF1a+/− and 10-HIF1a+/−. Entry probability was similar between 0-HIF1a+/− and 10-HIF1a+/− (p = 0.21); *p 0.05. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32493757), licensed under a CC-BY license. Not internally tested by R&D Systems.

The DU145FP cells express higher levels of cancer stemness related genes. (A) Transcriptome analyses of prostate cancer stemness-related markers. The Log2 transformed ratio of FPKM values (e.g., FP/WT) are indicated by color-coded index bars. (B) Cells were stained with fluorophore conjugated monoclonal antibodies against CD44 and EPCAM, the antibody against integrin alpha 2 beta 3 without fluorophore conjugation was detected by using secondary antibody conjugated with fluorophore, and stained cells were analyzed by flow cytometry. The scatter plots and histograms show representative results. CTRL denote cell stained with isotype control antibody or only secondary antibody; ST denote cell stained with antibody against EPCAM and integrin alpha 2 beta 3; US denote unstained cells. (C) Immunoblotting confirmation of the protein expression of A-tubulin, GAPDH, SOX9, SLUG, HIF1A, HIF2A, CD44 and integrin alpha 2 beta 3. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28698547), licensed under a CC-BY license. Not internally tested by R&D Systems.

Ten days of IH increases hippocampal HIF1a and disrupts Barnes maze performance in wild-type mice but not in HIF1a+/−. A, left, Representative digitized Western blotting images for HIF1a (103 kDa) and PCNA (40 kDa) in hippocampal nuclear protein fractions from control (n = 4) and IH10 (n = 4). Right, Quantification of HIF1a protein normalized to PCNA revealed that nuclear HIF1a was increased in IH10 when compared with control (p = 0.019). B, Total latency to exit the Barnes maze during three training sessions in control (n = 10) and in IH10 (n = 11). Each blue (control) and red (IH10) line represents an individual performance during training. Training to the exit was conducted over three sessions. Each session was separated by 24 hours. C, Left, During the probe trial, the distance traveled to initially enter the exit zone was shorter in control when compared with IH10 (p = 0.048). Right, Latency to initial entry was smaller in control as well (p = 0.034). D, Heat maps of the mean entry probability across all false exits (1–19) and the exit zone during probe trial for the control and IH10. Comparison of entry probability into the exit zone during the probe trial reveals that control has a greater probability for entering the exit zone when compared with IH10 (p = 0.004). E, Left, Representative digitized Western blotting images HIF1a and PCNA in hippocampal nuclear protein fractions from 0-HIF1a+/− (n = 4) and 10-HIF1a+/− (n = 4). Right, Quantification of HIF1a protein normalized to PCNA revealed that nuclear HIF1a is similar between 0-HIF1a+/− and 10-HIF1a+/− (p = 0.84). F, Total latency to exit the Barnes maze during three training sessions in 0-HIF1a+/− (n = 7) and in 10-HIF1a+/− (n = 8). Each gray (0-HIF1a+/−) and yellow (10-HIF1a+/−) line represents an individual performance during training. All experimental groups exhibit decreased total latency over the course of training. G, Left, In HIF1a+/−, the distance initial to initial entry into the exit zone was similar between 0-HIF1a+/− and 10-HIF1a+/− (p = 0.55). Right, Latency to initial entry into the exit zone during the probe trial were similar between 0-HIF1a+/− and 10-HIF1a+/− (p = 0.39). H, Heat maps of the mean entry probability into all zones during the probe trial for 0-HIF1a+/− and 10-HIF1a+/−. Entry probability was similar between 0-HIF1a+/− and 10-HIF1a+/− (p = 0.21); *p 0.05. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32493757), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu, Mu, Rt
• Applications: WB, Simple Western, IP, ChIP, ICC/IF, KO
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

HIF-1 alpha Antibody [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant human HIF-1 alpha /HIF1A
  Arg575-Asn826
  Accession # Q16665.1
• Specificity: Detects human, mouse and rat HIF-1 alpha /HIF1A.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: HIF1A
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Chromatin Immunoprecipitation (ChIP) 5 ug/5 x 10^6 cells
  • Immunocytochemistry 0.3-15 ug/mL
  • Immunoprecipitation 2 ug/500 ug cell lysate
  • Knockout Validated 0.3-15 ug/mL
  • Simple Western 5 ug/mL
  • Western Blot 0.5 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 HIF-1 alpha Antibody [Unconjugated]

• AINT
• anti-HIF-1 alpha
• anti-HIF1A
• ARNT interacting protein
• ARNT-interacting protein
• Basic-helix-loop-helix-PAS protein MOP1
• BHLHE78
• Class E basic helix-loop-helix protein 78
• HIF 1A
• HIF1 alpha
• HIF-1 alpha
• HIF1
• HIF1A
• HIF-1a
• HIF-1alpha
• HIF-1-alpha
• HIF1-alpha
• hypoxia inducible factor 1 alpha subunit, hypoxia inducible factor 1 subunit alpha
• hypoxia inducible factor 1, alpha subunit (basic helix-loop-helix transcription factor)
• hypoxia-inducible factor 1-alpha
• Member of PAS protein 1
• member of PAS superfamily 1
• MOP1
• PAS domain-containing protein 8
• PASD8

Background

The hypoxia-inducible transcription factor 1 alpha (HIF-1 alpha ) is the regulated member of the transcription factor heterodimer HIF-1. HIF-1 binds to hypoxia-response elements (HREs) in the promoters of many genes involved in adapting to an environment of insufficient oxygen or hypoxia. Hypoxic tissue environments occur in vascular and pulmonary diseases as well as cancer, which illustrates the broad impact of gene regulation by HIF-1 alpha .

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 HIF-1 alpha Antibody (AF1935)(47)

Publications using AF1935

• Sanmarco, LM;Rone, JM;Polonio, CM;Fernandez Lahore, G;Giovannoni, F;Ferrara, K;Gutierrez-Vazquez, C;Li, N;Sokolovska, A;Plasencia, A;Faust Akl, C;Nanda, P;Heck, ES;Li, Z;Lee, HG;Chao, CC;Rejano-Gordillo, CM;Fonseca-Castro, PH;Illouz, T;Linnerbauer, M;Kenison, JE;Barilla, RM;Farrenkopf, D;Stevens, NA;Piester, G;Chung, EN;Dailey, L;Kuchroo, VK;Hava, D;Wheeler, MA;Clish, C;Nowarski, R;Balsa, E;Lora, JM;Quintana, FJ; Lactate limits CNS autoimmunity by stabilizing HIF-1? in dendritic cells Nature 2023-08-09 [PMID: 37558878] (Immunohistochemistry, Mouse)
• Cornuault, L;Hérion, FX;Bourguignon, C;Rouault, P;Foussard, N;Alzieu, P;Chapouly, C;Gadeau, AP;Couffinhal, T;Renault, MA; Partial Mural Cell Ablation Disrupts Coronary Vasculature Integrity and Induces Systolic Dysfunction Journal of the American Heart Association 2023-06-22 [PMID: 37345826] (Immunohistochemistry, Transgenic Mouse)
• Yoshinori Aoki, Hongmei Dai, Fumika Furuta, Tomohisa Akamatsu, Takuya Oshima, Naoto Takahashi et al. LOX-1 mediates inflammatory activation of microglial cells through the p38-MAPK/NF-kappa B pathways under hypoxic-ischemic conditions Cell Communication and Signaling 6/2/2023 [PMID: 37268943]
• LM Sanmarco, JM Rone, CM Polonio, F Giovannoni, GF Lahore, K Ferrara, C Gutierrez-, N Li, A Sokolovska, A Plasencia, CF Akl, P Nanda, ES Heck, Z Li, HG Lee, CC Chao, CM Rejano-Gor, PH Fonseca-Ca, T Illouz, M Linnerbaue, JE Kenison, RM Barilla, D Farrenkopf, G Piester, L Dailey, VK Kuchroo, D Hava, MA Wheeler, C Clish, R Nowarski, E Balsa, JM Lora, FJ Quintana Engineered probiotics limit CNS autoimmunity by stabilizing HIF-1alpha in dendritic cells bioRxiv : the preprint server for biology, 2023-03-21;0(0):. 2023-03-21 [PMID: 36993446] (IHC, Mouse)
• Ioana V. Militaru, Alina Adriana Rus, Cristian V.A. Munteanu, Georgiana Manica, Stefana M. Petrescu New panel of biomarkers to discriminate between amelanotic and melanotic metastatic melanoma Frontiers in Oncology 1/26/2023 [PMID: 36776379]
• Ping Li, Dong-Ping Shi, Tao Jin, Dong Tang, Wei Wang, Liu-Hua Wang MTA1 aggravates experimental colitis in mice by promoting transcription factor HIF1A and up-regulating AQP4 expression Cell Death Discovery 6/28/2022 [PMID: 35764613]
• Eleanor R. Burgess, Rebekah L. I. Crake, Elisabeth Phillips, Helen R. Morrin, Janice A. Royds, Tania L. Slatter et al. Increased Ascorbate Content of Glioblastoma Is Associated With a Suppressed Hypoxic Response and Improved Patient Survival Frontiers in Oncology 3/28/2022 [PMID: 35419292]
• T Kjeld, AB Isbrand, K Linnet, B Zerahn, J Højberg, EG Hansen, LC Gormsen, J Bejder, T Krag, J Vissing, HE Bøtker, HC Arendrup Extreme Hypoxia Causing Brady-Arrythmias During Apnea in Elite Breath-Hold Divers Frontiers in Physiology, 2021-12-03;12(0):712573. 2021-12-03 [PMID: 34925050] (Western Blot, Human)
• G Zeng, T Wang, J Zhang, YJ Kang, L Feng FLI1 mediates the selective expression of hypoxia-inducible factor-1 target genes in endothelial cells under hypoxic conditions FEBS Open Bio, 2021-06-08;0(0):. 2021-06-08 [PMID: 34102031] (ICC, Western Blot, Human)
• X Fan, Z Zhao, J Song, D Zhang, F Wu, J Tu, M Xu, J Ji LncRNA-SNHG6 promotes the progression of hepatocellular carcinoma by targeting miR-6509-5p and HIF1A Cancer Cell International, 2021-03-04;21(1):150. 2021-03-04 [PMID: 33663502] (Western Blot, Human)
• Miao Liu, Nan Li, Chun Qu, Yilin Gao, Lijie Wu, Liangbiao George Hu Amylin deposition activates HIF1 alpha and 6-phosphofructo-2-kinase/fructose-2, 6-biphosphatase 3 (PFKFB3) signaling in failing hearts of non-human primates Communications Biology 2/12/2021 [PMID: 33580152]
• NC Balukoff, JJD Ho, PR Theodoridi, M Wang, M Bokros, LM Llanio, JR Krieger, JH Schatz, S Lee A translational program that suppresses metabolism to shield the genome Nat Commun, 2020-11-13;11(1):5755. 11/13/2020 [PMID: 33188200]
• B Di Marco, EE Crouch, B Shah, C Duman, MF Paredes, C Ruiz de Al, EJ Huang, J Alfonso Reciprocal Interaction between Vascular Filopodia and Neural Stem Cells Shapes Neurogenesis in the Ventral Telencephalon Cell Rep, 2020-10-13;33(2):108256. 2020-10-13 [PMID: 33053356] (IHC, Mouse)
• M Felsenstei, A Blank, AD Bungert, A Mueller, A Ghori, I Kremenetsk, O Rung, T Broggini, K Turkowski, L Scherschin, J Raggatz, P Vajkoczy, S Brandenbur CCR2 of Tumor Microenvironmental Cells Is a Relevant Modulator of Glioma Biology Cancers (Basel), 2020-07-13;12(7):. 2020-07-13 [PMID: 32668709] (IHC-F, Mouse)
• Arias-Cavieres A, Khuu M A et al. A HIF1a-Dependent Pro-Oxidant State Disrupts Synaptic Plasticity and Impairs Spatial Memory in Response to Intermittent Hypoxia. Eneuro 2024-06-19 [PMID: 32493757] (Simple Western, Mouse)
• Becker LM, O'Connell JT, Vo AP et al. Epigenetic Reprogramming of Cancer-Associated Fibroblasts Deregulates Glucose Metabolism and Facilitates Progression of Breast Cancer Cell Rep 6/2/2020 [PMID: 32492417]
• Ho JJD, Balukoff NC, Theodoridis PR et al. A network of RNA-binding proteins controls translation efficiency to activate anaerobic metabolism Nat Commun 5/29/2020 [PMID: 32472050]
  
  Details:
  Citation using the HRP format of this antibody.
• X Li, Y Zhao, C Chen, L Yang, HH Lee, Z Wang, N Zhang, MG Kolonin, Z An, X Ge, PE Scherer, K Sun The critical role of MMP14 in adipose tissue remodeling during obesity Mol. Cell. Biol., 2020-03-30;0(0):. 2020-03-30 [PMID: 31988105] (ChIP (chromatin immunoprecipit, Human)
• Cindy V Leiton, Elyssa Chen, Alissa Cutrone, Kristy Conn, Kennelia Mellanson, Dania M Malik et al. Astrocyte HIF-2α supports learning in a passive avoidance paradigm under hypoxic stress Hypoxia (Auckl) 12/7/2018 [PMID: 30519596]
• Michael A. Palladino, Genevieve A. Fasano, Dharm Patel, Christine Dugan, Marie London Effects of lipopolysaccharide-induced inflammation on hypoxia and inflammatory gene expression pathways of the rat testis Basic and Clinical Andrology 12/1/2018 [PMID: 30473791]
• K Tawara, H Scott, J Emathinger, A Ide, R Fox, D Greiner, D LaJoie, D Hedeen, M Nandakumar, AJ Oler, R Holzer, C Jorcyk Co-Expression of VEGF and IL-6 Family Cytokines is Associated with Decreased Survival in HER2 Negative Breast Cancer Patients: Subtype-Specific IL-6 Family Cytokine-Mediated VEGF Secretion Transl Oncol, 2018-11-12;12(2):245-255. 2018-11-12 [PMID: 30439625] (Western Blot, Human)
• AN Billin, SE Honeycutt, AV McDougal, JP Kerr, Z Chen, JM Freudenber, DK Rajpal, G Luo, HF Kramer, RS Geske, F Fang, B Yao, RV Clark, J Lepore, A Cobitz, R Miller, K Nosaka, AC Hinken, AJ Russell HIF prolyl hydroxylase inhibition protects skeletal muscle from eccentric contraction-induced injury Skelet Muscle, 2018-11-13;8(1):35. 2018-11-13 [PMID: 30424786] (Western Blot, Mouse)
• Q Yang, J Xu, Q Ma, Z Liu, V Sudhahar, Y Cao, L Wang, X Zeng, Y Zhou, M Zhang, Y Xu, Y Wang, NL Weintraub, C Zhang, T Fukai, C Wu, L Huang, Z Han, T Wang, DJ Fulton, M Hong, Y Huo PRKAA1/AMPK?1-driven glycolysis in endothelial cells exposed to disturbed flow protects against atherosclerosis Nat Commun, 2018-11-07;9(1):4667. 2018-11-07 [PMID: 30405100] (Western Blot, Human)
• Satoshi Sugita, Hideki Enokida, Hirofumi Yoshino, Kazutaka Miyamoto, Masaya Yonemori, Takashi Sakaguchi et al. HRAS as a potential therapeutic target of salirasib RAS inhibitor in bladder cancer International Journal of Oncology 6/11/2018 [PMID: 29901113]
• Z Liu, S Yan, J Wang, Y Xu, Y Wang, S Zhang, X Xu, Q Yang, X Zeng, Y Zhou, X Gu, S Lu, Z Fu, DJ Fulton, NL Weintraub, RB Caldwell, W Zhang, C Wu, XL Liu, JF Chen, A Ahmad, I Kaddour-Dj, M Al-Shabraw, Q Li, X Jiang, Y Sun, A Sodhi, L Smith, M Hong, Y Huo Endothelial adenosine A2a receptor-mediated glycolysis is essential for pathological retinal angiogenesis Nat Commun, 2017-09-19;8(1):584. 2017-09-19 [PMID: 28928465] (Western Blot, Human)
• Ying Xi, Thomas Kim, Alexis N. Brumwell, Ian H. Driver, Ying Wei, Victor Tan et al. Local lung hypoxia determines epithelial fate decisions during alveolar regeneration Nature Cell Biology 8/1/2017 [PMID: 28737769]
• L Li, R Zviti, C Ha, ZV Wang, JA Hill, F Lin Forkhead Box O3 (FoxO3) Regulates Kidney Tubular Autophagy Following Urinary Tract Obstruction J. Biol. Chem., 2017-07-13;0(0):. 2017-07-13 [PMID: 28705935] (Western Blot, Mouse)
• X Li, J Lu, Q Kan, X Li, Q Fan, Y Li, R Huang, A Slipicevic, HP Dong, L Eide, J Wang, H Zhang, V Berge, MA Goscinski, G Kvalheim, JM Nesland, Z Suo Metabolic reprogramming is associated with flavopiridol resistance in prostate cancer DU145 cells Sci Rep, 2017-07-11;7(1):5081. 2017-07-11 [PMID: 28698547] (Western Blot, Human)
• P Himmels, I Paredes, H Adler, A Karakatsan, R Luck, HH Marti, O Ermakova, E Rempel, ET Stoeckli, C Ruiz de Al Motor neurons control blood vessel patterning in the developing spinal cord Nat Commun, 2017-03-06;8(0):14583. 2017-03-06 [PMID: 28262664] (IHC, Mouse)
• Stacie K. Loftus, Laura L. Baxter, Julia C. Cronin, Temesgen D. Fufa, William J. NISC Comparative Sequencing Program, William J. NISC Comparative Sequencing Program Hypoxia-induced HIF1 alpha targets in melanocytes reveal a molecular profile associated with poor melanoma prognosis Pigment Cell & Melanoma Research 5/1/2017 [PMID: 28168807]
• Gabi U Dachs Ascorbate availability affects tumor implantation-take rate and increases tumor rejection in Gulo(-/-) mice Hypoxia (Auckl), 2016-04-08;4(0):41-52. 2016-04-08 [PMID: 27800507] (Western Blot, Mouse)
• Pandya PH, Wilkes DS. HIF-1alpha regulates CD55 expression in airway epithelium. Am J Respir Cell Mol Biol 8/6/2016 [PMID: 27494303]
• Ingrid Espinoza, Marcelo J. Sakiyama, Tangeng Ma, Logan Fair, Xinchun Zhou, Mohamed Hassan et al. Hypoxia on the Expression of Hepatoma Upregulated Protein in Prostate Cancer Cells Frontiers in Oncology 6/15/2016 [PMID: 27379206]
• P Storti, V Marchica, I Airoldi, G Donofrio, E Fiorini, V Ferri et al. Galectin-1 suppression delineates a new strategy to inhibit myeloma-induced angiogenesis and tumoral growth in vivo Leukemia 12/1/2016 [PMID: 27311934]
• Identification of ?-Dystrobrevin as a Direct Target of miR-143: Involvement in Early Stages of Neural Differentiation PLoS ONE, 2016-05-25;11(5):e0156325. 2016-05-25 [PMID: 27223470] (Western Blot, Human)
• HEREGULIN/ErbB3 SIGNALING ENHANCES CXCR4-DRIVEN Rac1 ACTIVATION AND BREAST CANCER CELL MOTILITY VIA HIF-1? Mol Cell Biol, 2016-07-14;0(0):. 2016-07-14 [PMID: 27185877] (ChIP, Human)
• Zuo R, Gu X, Qi Q, Wang T, Zhao X, Liu J, Yang Z Warburg-like Glycolysis and Lactate Shuttle in Mouse Decidua during Early Pregnancy. J Biol Chem, 2015-07-15;290(35):21280-91. 2015-07-15 [PMID: 26178372] (Western Blot, Mouse)
• Li R, Uttarwar L, Gao B, Charbonneau M, Shi Y, Chan J, Dubois C, Krepinsky J High Glucose Up-regulates ADAM17 through HIF-1alpha in Mesangial Cells. J Biol Chem, 2015-07-14;290(35):21603-14. 2015-07-14 [PMID: 26175156] (ChIP, Rat)
• Rosa Paolillo, Isabella Spinello, Maria Teresa Quaranta, Luca Pasquini, Elvira Pelosi, Francesco Lo Coco et al. Human TM9SF4 Is a New Gene Down-Regulated by Hypoxia and Involved in Cell Adhesion of Leukemic Cells PLOS ONE 5/11/2015 [PMID: 25961573]
• Tholen M, Wolanski J, Stolze B, Chiabudini M, Gajda M, Bronsert P, Stickeler E, Rospert S, Reinheckel T Stress-resistant Translation of Cathepsin L mRNA in Breast Cancer Progression. J Biol Chem, 2015-05-08;290(25):15758-69. 2015-05-08 [PMID: 25957406] (Western Blot, Human)
• Palladino M, Shah A, Tyson R, Horvath J, Dugan C, Karpodinis M Myeloid cell leukemia-1 (Mc1-1) is a candidate target gene of hypoxia-inducible factor-1 (HIF-1) in the testis. Reprod Biol Endocrinol, 2012-12-05;10(0):104. 2012-12-05 [PMID: 23216940] (ChIP, Direct ELISA, Mouse)
• Liang D, Ma Y, Liu J, Trope CG, Holm R, Nesland JM, Suo Z The hypoxic microenvironment upgrades stem-like properties of ovarian cancer cells. BMC Cancer, 2012-05-29;12(0):201. 2012-05-29 [PMID: 22642602] (Western Blot, Human)
• Robador PA, San Jose G, Rodriguez C, Guadall A, Moreno MU, Beaumont J, Fortuno A, Diez J, Martinez-Gonzalez J, Zalba G HIF-1-mediated up-regulation of cardiotrophin-1 is involved in the survival response of cardiomyocytes to hypoxia. Cardiovasc. Res., 2011-07-19;92(2):247-55. 2011-07-19 [PMID: 21771897] (ChIP, Human)
• Joshi D, Patel H, Baker DM, Shiwen X, Abraham DJ, Tsui JC Development of an in vitro model of myotube ischemia. Lab. Invest., 2011-05-23;91(8):1241-52. 2011-05-23 [PMID: 21606923] (Western Blot, Mouse)
• Bollig-Fischer A, Dziubinski M, Boyer A, Haddad R, Giroux CN, Ethier SP HER-2 signaling, acquisition of growth factor independence, and regulation of biological networks associated with cell transformation. Cancer Res., 2010-08-24;70(20):7862-73. 2010-08-24 [PMID: 20736364] (Western Blot, Human)
• Wang B, Wood IS, Trayhurn P Hypoxia induces leptin gene expression and secretion in human preadipocytes: differential effects of hypoxia on adipokine expression by preadipocytes. J. Endocrinol., 2008-05-07;198(1):127-34. 2008-05-07 [PMID: 18463145] (Western Blot, Human)
• Colla S, Tagliaferri S, Morandi F, Lunghi P, Donofrio G, Martorana D, Mancini C, Lazzaretti M, Mazzera L, Ravanetti L, Bonomini S, Ferrari L, Miranda C, Ladetto M, Neri TM, Neri A, Greco A, Mangoni M, Bonati A, Rizzoli V, Giuliani N The new tumor-suppressor gene inhibitor of growth family member 4 (ING4) regulates the production of proangiogenic molecules by myeloma cells and suppresses hypoxia-inducible factor-1 alpha (HIF-1alpha) activity: involvement in myeloma-induced angiogenesis. Blood, 2007-09-11;110(13):4464-75. 2007-09-11 [PMID: 17848618] (Western Blot, Human)

Reviews for HIF-1 alpha Antibody (AF1935) (2)

Average Rating: 4

(Based on 2 reviews)

We have 2 reviews tested in 2 species: Mouse, Mouse and mouse brain Tissue.

reviewed by: Bhavin Shah IHC Mouse and mouse brain Tissue 10/29/2018

Summary

• Application: Immunohistochemistry
• Sample Tested: 80µm vibratome sections of P12 mouse brain cortex,PFA fixed P12 mouse brain Cortex-80µm vibratome
• Species: Mouse and mouse brain Tissue
• Lot: KAK0214121

Comments

• Comments: Perkin Elmer, antigen-antibody signal amplification Kit. Immunohistochemistry, vibratome sections of 80µm of PFA fixed P12 mouse brain cortex. Can also be used for Frozen PFA fixed sections

reviewed by: Verified Customer WB Mouse 01/05/2015

Summary

• Application: Western Blot
• Sample Tested: See PMIC 23918831
• Species: Mouse

FAQs for HIF-1 alpha Antibody (AF1935). (Showing 1 - 10 of 11 FAQs).

1. Why is there a difference between the theoretical MW for HIF1A and the observed MW for HIF-1 alpha?
   • HIF1A, like many other proteins, has post-translational modifications. Depending on the size, amount and nature of the post-translational modifications, it can cause subtle to very large changes in molecular weight.
2. Which antibody(ies) do you recommend for the detection of HIF-1a by immunohistochemistry in the sections of paraffin-embedded mouse liver samples? I would appreciate if you can give me several choices and rank them in the order of performance. My goal is to distinguish HIF upregulation by prolyl hydroxylase inhibitor in different liver cells.
   • All of our antibodies are of high quality and are well tested/validated in species/applications we list on the datasheet. However, we suggest the following four HIF-1 alpha antibodies based upon customer reviews, as well as the number of peer reviewed publications in which these products have been cited by researchers from reputed institutes. (1) HIF-1 alpha Antibody (H1alpha67) (cat# NB100-105) (cited in at least 218 peer reviewed publications) (2) HIF-1 alpha Antibody (cat# NB100-479) (cited in at least 51 peer reviewed publications) (3) HIF-1 alpha Antibody (H1alpha67) (cat# NB100-123 ) (cited in at least 38 peer reviewed publications) (4) HIF-1 alpha Antibody (cat# NB100-449) (cited in at least 31 peer reviewed publications).
3. I would like to know, does a path exist for detection of HIF 1 in venous blood before and after revascularization of the leg? 
   • We are not entirely sure if HIF-1 alpha will be present in the leg after revascularization. It may be present, but you may want to search the literature to see if this has been looked at before. If not, then this would certainly be an experiment worth doing.
4. What is the molecular weight (kDa) of protein HIF 1 alpha in western blot?
   • The theoretical molecular weight of HIF 1-alpha is ~93kDa. However, you will likely see a band between 100-120kDa due to phosphorylation.
5. We got the Hif1a (NB100-105) antibody from you guys. I used the concentration that is mentioned on your website, but I am getting a band of a completely different size (~70kDa) and not the 120 kDa mentioned.
   • HIF-1 alpha is a notoriously difficult protein to work with due to its rapid degradation. Therefore, the ~70kDa bands are most likely degradation products. It is very important to lyse the cells in hypoxic conditions. We strongly recommend lysing the cells directly into the Laemmli buffer and doing that quickly, so that the exposure to oxygen is minimized.Please go through our hypoxia related FAQs, you should find them very informative.Also, running a positive control may help confirm the band specificity in your samples. You may prepare them yourself or choose some from our catalog, for example: 1) HeLa Hypoxic / Normoxic Cell Lysate (NBP2-36452)2) HeLa Hypoxic (CoCl2) / Normoxic Cell Lysate (NBP2-36450)
6. I performed several Western Blots of HIF-1 alpha with different lysis buffers, whole lysates, and cytoplasm/nuclei extractions. I can’t seem to get a good western blot (poor signal, band much lower than expected, etc.). Can someone suggest some technical considerations/tricks I should consider using?
   • A major issue that researchers working with HIF-1 alpha is degradation due to exposure to oxygen. In western blot, this results in a weaker band and/or the appearance of multiple low molecular weight bands (40-80 kDa). We recommend preparing the lysates after collection of cells/tissues as quickly as possible (on ice), preferably in a hypoxic chamber. We also recommend including a true hypoxia mimetic control (eg: cells treated with CoCl2, DMOG… etc.). The controls help distinguish your band of interest from potential degradation/dimer bands.For more troubleshooting tips and frequently asked questions regarding hypoxia/HIFs, you can refer to our hypoxia-related FAQs.
7. I am doing HIF1 westerns in HIF-overexpressing mouse liver and adipose tissue using Novus antirabbit HIF1a antibody with overnight incubation. I am getting strong bands around 90kDa. I am aware that HIF theoretical molecular weight is 93kDa, but in westerns, the HIF band is usually around 120kDa according to my internet research. Can someone let me know if I’m getting the right HIF band or just some non-specific bands? Thanks.
   • (1)    HIF-1 alpha’s theoretical molecular weight is 93kDa. The post translationally modified/ubiquitinated form of HIF-1 alpha protein (fails to undergo proteasomal degradation) shows up as a band in the 110-130 kDa range on a Western blot.(2)    The dimeric protein may appear at a position above 200 kDa on non-reducing gels.(3)    Importantly, HIFs are among the most rapidly degradable proteins; therefore, sample preparation is highly important when analyzing HIF1 alpha or HIF2 alpha. When degraded, HIF-1 alpha may show up between 40-80 kDa position on Western blot. Degradation may be avoided by preparing the samples as soon as possible after collection of cells/tissues in hypoxic chamber. Notably, the tissues/cells should be kept on ice during lysate preparation and the lysates should be analyzed as soon as possible.(4)    For troubleshooting suggestions/feedback on more than 25 similar frequently asked questions, I would recommend visiting Novus page: FAQs - Hypoxia and HIFs (5)    Last but not the least, Novus technical support team may be contacted via email
8. I have Hif1a nuclear protein extract at -80C. I am wondering if anyone knows how long it would be good for at that temperature since HIf1a is known to be degraded easily.Thank you!
   • You could try a few things to further inhibit the degradation.1) Use the protease inhibitors (if you are not already using them).2) Lyse cells into a buffer that contains SDS or LDS (eg: Laemmli's buffer), since SDS and LDS denature and inhibit proteases. Lysis may even be performed with reducing agents in the buffer (eg. DTT), but this will make your lysates unsuitable for BCA assay.3) Lysing samples rapidly ensures that the samples are instantly homogenized (it also shears DNA released by the SDS).5) Flash-freezing samples in liquid nitrogen rather than freezing at -80*C reduces the window of time for protease activity.6) Freeze samples in individual aliquots, instead of thawing the same vial multiple times.
9. I am curious to know the biochemical reactions of CoCl2 that mimic hypoxia. Is it that CoCl2 can bind any ubiquitin enzyme which regulates their degradation?
   • CoCl2 inhibits PHD enzymes (the body’s “oxygen sensors”) by replacing the Fe ion with Co, preventing these enzymes from marking HIF-1 alpha for degradation. CoCl2-based hypoxia mimetic samples are often used as positive control in HIF analysis. For more troubleshooting tips and frequently asked questions regarding hypoxia/HIFs, you can refer to our hypoxia-related FAQs.
10. I am curious to know the biochemical reactions of CoCl2 that mimic hypoxia. Is it that CoCl2 can bind any ubiquitin enzyme which regulates their degradation?
    • CoCl2 inhibits PHD enzymes (the body’s “oxygen sensors”) by replacing the Fe ion with Co, preventing these enzymes from marking HIF-1 alpha for degradation. CoCl2-based hypoxia mimetic samples are often used as positive control in HIF analysis. For more troubleshooting tips and frequently asked questions regarding hypoxia/HIFs, you can refer to our hypoxia-related FAQs.
11. Is cross-reactivity with HIF-2 alpha tested/predicted?
    • Although we don’t have cross-reactivity data with regards to HIF-2 alpha, we predict minimal cross-reactivity based on low sequence similarity observed from BLAST analysis between HIF-1 alpha and HIF-2 alpha.
Show All 11 FAQs.

Bioinformatics

• Gene Symbol: HIF1A
• Uniprot:
  • Q16665.1()