PA3-16521

antibody from Invitrogen Antibodies

Targeting: HIF1A bHLHe78, HIF-1alpha, HIF1, MOP1, PASD8

Western blot (Data presented on provider website)

ELISA (Recommended by provider)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunoprecipitation (Recommended by provider)

Immunohistochemistry (Supportive data in Antibodypedia)

Gel shift (Recommended by provider)

Chromatin Immunoprecipitation (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA3-16521

Provider

Invitrogen Antibodies

Product name

HIF1A Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Other

Description

For ChIP, nuclear extracts are recommended. Prior to immunostaining paraffin tissues, antigen retrieval with sodium citrate buffer (pH 6.0) is recommended. Suggested positive control: Cos-7 Hypoxia Induced nuclear extract Kit6.

Reactivity

Human, Mouse, Rat, Bovine, Canine, Guinea Pig, Xenopus, Zebrafish

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Concentration

1 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

References

Submitted references

α-Lipoic Acid Maintains Brain Glucose Metabolism via BDNF/TrkB/HIF-1α Signaling Pathway in P301S Mice.

Zhang YH, Yan XZ, Xu SF, Pang ZQ, Li LB, Yang Y, Fan YG, Wang Z, Yu X, Guo C, Ao Q
Frontiers in aging neuroscience 2020;12:262

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Nox2 and Cyclosporine-Induced Renal Hypoxia.

Djamali A, Wilson NA, Sadowski EA, Zha W, Niles D, Hafez O, Dorn JR, Mehner TR, Grimm PC, Hoffmann FM, Zhong W, Fain SB, Reese SR
Transplantation 2016 Jun;100(6):1198-210

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Dihydrotestosterone attenuates hypoxia inducible factor-1α and cyclooxygenase-2 in cerebral arteries during hypoxia or hypoxia with glucose deprivation.

Zuloaga KL, Gonzales RJ
American journal of physiology. Heart and circulatory physiology 2011 Nov;301(5):H1882-90

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Mouse snail is a target gene for HIF.

Luo D, Wang J, Li J, Post M
Molecular cancer research : MCR 2011 Feb;9(2):234-45

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of HIF1A was performed using 70% confluent log phase MCF7 cells treated with Cobalt chloride (150 µM for 48hrs). The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with HIF1A Polyclonal Antibody (Product # PA3-16521) at 1:200 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:3000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nucleus localization in CoCl2 treated MCF7 cells. Panel e represents no staining observed for untreated MCF7 cells. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical analysis of HIF1A in human kidney, renal tubular epithelium in cortex. Samples were incubated in HIF1A polyclonal antibody (Product # PA3-16521).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical analysis of HIF1A in formalin-fixed paraffin-embedded tissue section of human endometrium carcinoma AN3CA cell line based xenograft. Samples were incubated in HIF1A polyclonal antibody (Product # PA3-16521) using a dilution of 1:300 followed by a HRP-labeled secondary antibody and DAB reagent. 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).

Chromatin Immunoprecipitation

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Chromatin Immunoprecipitation (ChIP) assay of endogenous HIF1A was performed using HIF1A Polyclonal Antibody (Product # PA3-16521, 5 µg) on sheared chromatin from MCF-7 cells treated with Cobalt Chloride (150 µM for 48hrs) using the MAGnify ChIP System kit (Product # 49-2024). Chromatin from untreated MCF-7 cells were prepared the same way. Normal Rabbit IgG was used as a negative IP control. The purified DNA was analyzed by qPCR using primers binding to VEGFA-PR, ARRDC3, ERRF1-site 1 (active) and ERRF1-site 2 and SAT alpha (inactive). Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

NULL

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 3 LA mediated the upregulation of hypoxia-inducible factor-1alpha (HIF-1alpha) expression via brain-derived neurotrophic factor/tyrosine Kinase receptor B (BDNF/TrkB) pathway. (A) Western blot results showed the protein expression levels of nuclear HIF-1alpha, Lamin, and cytosol HIF-1alpha, BDNF, p-TrkA/B, TrkB. (B) Quantified results of nuclear HIF-1alpha levels. Lamin served as an internal loading control. (C-F) Relative protein levels of cytosol HIF-1alpha, BDNF, p-TrkB, TrkB-FL which were established after normalization to beta-actin. (G,H) mRNA levels of HIF-1alpha and BDNF. Values are represented as the means +- SEM ( n = 7). * p < 0.05, ** p < 0.01 compared with the vehicle group.