MA1-16517

antibody from Invitrogen Antibodies

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

Western blot (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

MA1-16517

Provider

Invitrogen Antibodies

Product name

HIF1A Monoclonal Antibody (H1alpha 67-7)

Antibody type

Monoclonal

Antigen

Other

Description

In Western blot, multiple bands may be seen at 120 kDa representing post-translational modification of HIF-1 alpha. Nuclear extracts are recommended. Suggested positive control: Cos7 CoCl2-treated nuclear extract.

Reactivity

Human

Host

Mouse

Isotype

IgG

Antibody clone number

H1alpha 67-7

Vial size

100 µL

Concentration

1.5 mg/mL

Storage

4° C, do not freeze

References

Submitted references

Intrabody Targeting HIF-1α Mediates Transcriptional Downregulation of Target Genes Related to Solid Tumors.

Hu Y, Romão E, Vincke C, Brys L, Elkrim Y, Vandevenne M, Liu C, Muyldermans S
International journal of molecular sciences 2021 Nov 15;22(22)

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of HIF1A in 1: Cos7 CoCl2 treated cells. 2: Untreated cells. Samples were incubated in HIF1A monoclonal antibody (Product # MA1-16517).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-HIF1A Monoclonal Antibody (H1alpha 67-7) (Product # MA1-16517) and a 92kDa band corresponding to Hypoxia-inducible factor 1-alpha was observed in Hela upon Cobalt chloride treatment. Nuclear enriched extracts (50 µg lysate) of HeLa (Lane 1) and HeLa treated with Cobalt chloride (150uM for 48hr) (Lane 2) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 5 Immunoprecipitation of HIF-1alpha protein by selected Nbs. HeLa cells stimulated with CoCl 2 were harvested and lysed with freeze-thaw cycles. The cell lysate was collected and incubated with Nbs to capture HIF-1alpha protein. Protein A/G coated agarose beads were incubated with monoclonal anti-His tag IgG to precipitate the Nbs from HeLa cell lysate. The precipitation was loaded on the SDS-PAGE and transferred onto a NC membrane for detection of the native HIF-1alpha via Western blot with mouse anti-HIF-1alpha IgG and HRP conjugated anti-mouse IgG. Immunoprecipitation with mouse anti-HIF-1alpha IgG instead of Nbs was performed and served as the positive control. Immunoprecipitation with mouse anti-His tag IgG instead of Nbs was performed and served as the negative control. The light and heavy chains from mouse anti-His IgG were indicated as 'L-chain' and 'H-chain', respectively.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 4 Determination of the condition to stabilize endogenous HIF-1alpha protein in HeLa cells via Western blot. HeLa cells were stimulated with the condition indicated on the figure or left untreated. Then, cells were collected and lysed by freeze-thaw cycles. The released cytoplasm was separated by SDS-PAGE and transferred to a nitrocellulose membrane for Western blot analysis. Mouse anti-HIF-1alpha IgG and anti-tubulin IgG were incubated with the membranes and HRP conjugated anti-mouse IgG was used to confirm the presence of the bands. The statistical difference in the figures is indicated as follows: * ( p < 0.05), **** ( p < 0.0001).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 5 Immunoprecipitation of HIF-1alpha protein by selected Nbs. HeLa cells stimulated with CoCl 2 were harvested and lysed with freeze-thaw cycles. The cell lysate was collected and incubated with Nbs to capture HIF-1alpha protein. Protein A/G coated agarose beads were incubated with monoclonal anti-His tag IgG to precipitate the Nbs from HeLa cell lysate. The precipitation was loaded on the SDS-PAGE and transferred onto a NC membrane for detection of the native HIF-1alpha via Western blot with mouse anti-HIF-1alpha IgG and HRP conjugated anti-mouse IgG. Immunoprecipitation with mouse anti-HIF-1alpha IgG instead of Nbs was performed and served as the positive control. Immunoprecipitation with mouse anti-His tag IgG instead of Nbs was performed and served as the negative control. The light and heavy chains from mouse anti-His IgG were indicated as 'L-chain' and 'H-chain', respectively.