MA1-16504

Product number: MA1-16504 - Provider product page
Provider: Invitrogen Antibodies
Product name: HIF1A Monoclonal Antibody (H1alpha67)
Antibody type: Monoclonal
Antigen: Other
Description: In Western blot, multiple bands may be seen at 100-120 kDa representing post-translational modification of HIF-1 alpha. For WB, testing on nuclear extracts is recommended. Recommended positive controls: COS-7 nuclear hypoxic induced cell lysate, HeLa hypoxic (CoCl2) cell lysate, HepG2 hypoxic (CoCl2) cell lysate, HeLa hypoxic cell lysate, HepG2 hypoxic cell lysate, HIF-1 alpha Knockout HeLa cell lysate
Reactivity: Human, Mouse, Rat, Bovine, Canine, Feline, Porcine, Rabbit, Xenopus
Host: Mouse
Isotype: IgG
Antibody clone number: H1alpha67
Vial size: 100 µL
Concentration: 1.0 mg/mL
Storage: Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

HIF1A protein structure - MA1-16504 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 6 D-Lactate induces HIF-1alpha protein accumulation under normoxic conditions in bFLS. bFLS were stimulated with 5 mM D-lactate for 6 h under normoxic (20% O 2 ) and hypoxic (1% O 2 ) conditions. bTNF-alpha was used as the positive control. HIF-1alpha stabilization was detected in total protein extracts by Western blot using cobalt chloride (CoCl 2 ) as the hypoxia-mimetic agent. (A) Representative HIF-1alpha immunoblot is shown. (B) Densitometry HIF-1alpha values were quantified using Image Studio Lite v5.2 software and normalized to beta-actin. Each bar represents the mean +- SEM, n = 3. ** p < 0.01; **** p < 0.0001; n.s ., not significant.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: FIGURE 1 Effects of oxygen concentration on growth, cell-death, gene expression and metabolism in human colonoids (A) . Experimental design. Differentiated colonoids from six donors were either grown continuously at 20% O 2 or calibrated at 2% O 2 for 16h before treated with TNF (100ngml -1 ), IL17 (100ngml -1 ) or a combination (TNF/IL17) for 24h (B) Detection of morphology and cell death by live imaging of untreated colonoids. Left image shows a 3D depth encoded image of one colonoid at 20% O 2 , according to the color scale from red (superficial) to blue (deep) on the right side of the image. Center and right image panels show colonoids in 20% or 2% O 2 respectively, where NucBlue (r) Live reagent (Hoechst 33342) stains all cell nuclei (blue) and propidium iodide stains dead cell nuclei (red). Scale bars 100um (C) Expression of epithelial cell markers as measured by flow cytometry in untreated colonoids from three donors, in 20% O 2 or 2% O 2 as depicted in panel (A) ( n = 3 + 3 for each marker). Relative change between oxygen concentrations normalized to 20% O 2 (red line at y = 1) displayed, with mean and individual normalized values (D) and (E) Immunostaining of differentiated colonoids grown at 20% or 2% O 2 , representative selection from five assays. Scale bars 50um. (D) Ki67 positive nuclei stained brown, counterstaining with hematoxylin. The graph shows percentage of Ki67 positive nuclei in four independent experiments indicated by dots. Bars show mean with SD (E)

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 4 Immunohistochemical evaluation of letrozole-treated chorioallantoises (CA). ( A ) Immunohistochemical demonstration of capillary densities in chorioallantoic membranes of letrozole-treated and control mares using vWF. The capillary density tended to be greater in the control CAs than in the letrozole-treated CAs, p = 0.07; ( B ) A strong cytoplasmic signal for VEGF is noted within the chorionic epithelium of letrozole-treated group. No difference was observed in the abundance of HIF1A + cells between the groups. ( B ) Red boxes (letrozole) and blue circles (control) represent values for the capillary density scores in the chorioallantoic samples.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 1 Expression analysis of HIF1A in granulosa cells. ( a ) indicates the mRNA expression of HIF1A under different FSH and IGF1 concentrations. ( b ) Indicates the detection of HIF1A protein under normoxia. The lane numbers 1-4 indicate the western runs of individual protein lysates. Columns 1 and 2 represent duplicates of HIF1A probing in FSH (20 ng/mL) and IGF1(50 ng/mL) treated GC under normoxia while columns 3 and 4 represent the Beta actin (BACT) probing in the corresponding samples. The arrow marks in ( b ) indicate the HIF1A (columns 1 and 2) and BACT (columns 3 and 4) protein bands. The image is obtained with the exposure time of 8 seconds in Simple Western instrument. ( c ) illustrates the immunolocalization of HIF1A in different growing ovarian follicles of abattoir ovaries. The brown color immunochemical signal indicates the HIF1A staining inside the follicle. 400x optical magnification was used for primary and secondary and large antral follicle's wall. 100x optical magnification was used for tertiary follicles in the pictures. Inside dotted box indicates the specific follicle for primary, secondary and tertiary follicle sections whereas dotted box points the GC layer in large antral follicle's wall. Data were presented in Mean +- SEM values of three independent cell culture experiments (n = 3) for Fig. 1a. Significant differences were acknowledged at the minimum level of p < 0.05 by one way repeated measures analysis of variance. Pairwise comparisons were ana

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 8 Chromatin immunoprecipitation analysis. ( a ) shows the nucleotide sequence between the physical location of 59122186 to 59122473 nucleotides on chromosome 10 in the bovine genome. Manually curated TATA box and the putative HIF1 binding sites were highlighted in blue and yellow colors. The primer binding sites were indicated with an underline. ( b ) adopted JASPAR Sequence logo of the HIF1 binding site based on ChIP seq data in humans. ( c ) indicates the agarose gel pictures of immune-precipitated and input DNA samples. ( d , e ) depict the relative quantification of HIF1 binding events on the CYP19A1 promoter based on percent input and fold change methods, respectively under FSH and IGF1 Supplemented conditions. Data were represented in mean +- SEM values of three independent experiments (n = 3). Significant differences were acknowledged at the minimum level of p < 0.05 by one way repeated measures analysis of variance. Pairwise comparisons were analyzed using Post hoc Tuckey test.