Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human, Mouse, Rat, Transgenic Mouse

Applications

Validated:

Knockout Validated, Western Blot, Immunocytochemistry, Simple Western, Immunoprecipitation, Chromatin Immunoprecipitation (ChIP)

Cited:

Immunohistochemistry, Western Blot, Direct ELISA, Immunocytochemistry, Simple Western, Chromatin Immunoprecipitation (ChIP), IHC-F

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all HIF-1 alpha/HIF1A Primary Antibodies »

Product Specifications

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.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human/Mouse/Rat HIF-1 alpha/HIF1A Antibody

Detection of Human HIF-1a/HIF1A antibody by Western Blot.

Detection of Human HIF‑1 alpha /HIF1A by Western Blot.

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.
Detection of HIF-1a/HIF1A-regulated Genes antibody by Chromatin Immunoprecipitation.

Detection of HIF‑1 alpha /HIF1A-regulated Genes by Chromatin Immunoprecipitation.

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-1a/DNA complexes were immunoprecipitated using 5 µg Goat Anti-Human/Mouse/Rat HIF-1a/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 Antibody Specificity is Shown by Immunocytochemistry in Knockout Cell Line by Immunocytochemistry (ICC).

HIF‑1 alpha /HIF1A Specificity is Shown by Immunocytochemistry in Knockout Cell Line.

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.
Detection of Human HIF-1a/HIF1A antibody by Simple WesternTM.

Detection of Human HIF‑1 alpha /HIF1A by Simple WesternTM.

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-1a/HIF1A at approximately 115 kDa (as indicated) using 5 µg/mL of Goat Anti-Human/Mouse/Rat HIF-1a/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 the 12-230 kDa separation system.
Detection of Mouse HIF-1 alpha by Simple Western

Detection of Mouse HIF-1 alpha by Simple Western

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; **p < 0.01; N.S., p > 0.05. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/32493757), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human HIF-1 alpha by Western Blot

Detection of Human HIF-1 alpha by Western Blot

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 (https://pubmed.ncbi.nlm.nih.gov/28698547), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse HIF-1 alpha by Simple Western

Detection of Mouse HIF-1 alpha by Simple Western

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; **p < 0.01; N.S., p > 0.05. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/32493757), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human HIF-1 alpha/HIF1A by Immunocytochemistry/ Immunofluorescence

Detection of Human HIF-1 alpha/HIF1A by Immunocytochemistry/ Immunofluorescence

Staining of HIF1A in HIF1A KO and WT HUVECs. (a–d) One representative image of the stained HIF1A KO and WT HUVECs, which were stained with Hoechst (blue), anti-actin (green), and anti-HIF1A (red). (a) WT HUVECs cultured in normoxic conditions. (b) WT HUVECs cultured in hypoxic conditions. (c) HIF1A KO HUVECs cultured in normoxic conditions. (d) HIF1A KO HUVECs cultured in hypoxic conditions. (e) Quantification of the HIF1A signal intensity (MFI) in the nucleus and the cytoplasm in WT and KO HUVECs in normoxic and hypoxic conditions, which is compared by two-way ANOVA multiple comparisons test, *** p ≤ 0.001. The experiment was performed with one biological replicate at p. 3 (n = 3) and five images were taken per well. Scale bar 10 µm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36671408), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human HIF-1 alpha/HIF1A by Immunocytochemistry/ Immunofluorescence

Detection of Human HIF-1 alpha/HIF1A by Immunocytochemistry/ Immunofluorescence

Staining of HIF1A in HIF1A KO and WT HUVECs. (a–d) One representative image of the stained HIF1A KO and WT HUVECs, which were stained with Hoechst (blue), anti-actin (green), and anti-HIF1A (red). (a) WT HUVECs cultured in normoxic conditions. (b) WT HUVECs cultured in hypoxic conditions. (c) HIF1A KO HUVECs cultured in normoxic conditions. (d) HIF1A KO HUVECs cultured in hypoxic conditions. (e) Quantification of the HIF1A signal intensity (MFI) in the nucleus and the cytoplasm in WT and KO HUVECs in normoxic and hypoxic conditions, which is compared by two-way ANOVA multiple comparisons test, *** p ≤ 0.001. The experiment was performed with one biological replicate at p. 3 (n = 3) and five images were taken per well. Scale bar 10 µm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36671408), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human/Mouse/Rat HIF-1 alpha/HIF1A Antibody

Application
Recommended Usage

Chromatin Immunoprecipitation (ChIP)

5 µg/5 x 106 cells
Sample: CoCl2 treated mouse primary kidney cells chromatin,  epo promoter detected by standard PCR.

Immunocytochemistry

0.3-15 µg/mL
Sample: Immersion fixed HeLa human cervical epithelial carcinoma cell line treated with DFO

Immunoprecipitation

2 µg/500 µg cell lysate
Sample: MCF‑7 human breast cancer cell line treated with CoCl2, see our available Western blot detection antibodies

Knockout Validated

HIF-1 alpha /HIF1A is specifically detected in HeLa human cervical epithelial carcinoma cell line but is not detectable in HIF-1 alpha/HIF1A knockout HeLa cell line.

Simple Western

5 µg/mL
Sample: A549 human lung carcinoma cell line treated with Hypoxia (1% O2),

Western Blot

0.5 µg/mL
Sample: CoCl2-treated MCF‑7 human breast cancer cell line

Reviewed Applications

Read 3 reviews rated 4 using AF1935 in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

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

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: HIF-1 alpha/HIF1A

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.

Long Name

Hypoxia Inducible Factor 1 Subunit Alpha

Alternate Names

BHLHE78, HIF 1A, HIF-1a, HIF1 alpha, HIF1A, MOP1, PASD8

Entrez Gene IDs

3091 (Human); 15251 (Mouse); 29560 (Rat)

Gene Symbol

HIF1A

UniProt

Q16665.1

Additional HIF-1 alpha/HIF1A Products

Product Documents for Human/Mouse/Rat HIF-1 alpha/HIF1A Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human/Mouse/Rat HIF-1 alpha/HIF1A Antibody

For research use only

Citations for Human/Mouse/Rat HIF-1 alpha/HIF1A Antibody

Customer Reviews for Human/Mouse/Rat HIF-1 alpha/HIF1A Antibody (3)

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3 Customer Ratings
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  • Human/Mouse/Rat HIF-1 alpha/HIF1A Antibody
    Name: Anonymous
    Sample Tested: HASMC human aortic smooth muscle cells
    Species: Human
    Verified Customer | Posted 12/04/2019
    I have tried HIF1A antibodies from several companies, this one is good compared to others, but sometimes there are multiple bands. 1:500
    Human/Mouse/Rat HIF-1 alpha/HIF1A Antibody AF1935
  • Human/Mouse/Rat HIF-1 alpha Antibody
    Name: Bhavin Shah
    Application: Immunohistochemistry
    Sample Tested: PFA fixed P12 mouse brain Cortex-80µm vibratome and 80µm vibratome sections of P12 mouse brain cortex
    Species: mouse brain Tissue and Mouse
    Verified Customer | Posted 10/29/2018
    HIF1a stained brain section -Biotin-streptavidin as secondary- the secondary antibody signal was amplified using Perkin Elmer amplification kit. The P12 section shows upper layer of neurons as Hif1a positive
    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
    Human/Mouse/Rat HIF-1 alpha/HIF1A Antibody AF1935
  • Name: Anonymous
    Application: Western Blot
    Sample Tested: See PMIC 23918831
    Species: Mouse
    Verified Customer | Posted 01/05/2015

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