PA1-16627

antibody from Invitrogen Antibodies

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

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Gel shift (Recommended by provider)

Antibody Data

Product number

PA1-16627

Provider

Invitrogen Antibodies

Product name

HIF1A Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Other

Description

This antibody has 93% sequence homology with bovine and 90% sequence homology with mouse. Suggested positive control: Cos-7 Hypoxia induced nuclear extract, COS-7 nuclear extracts (treated).

Reactivity

Human, Mouse, Rat, Bovine, Porcine

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Concentration

1 mg/mL

Storage

-20° C, Avoid Freeze/Thaw Cycles

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot detection of HIF-1 alpha in Cobalt Chloride treated/untreated Cos-7 nuclear extracts using Product # PA1-16627.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of HIF1A in 0.5 mg/mL Hypoxic HeLa lysate. Samples were incubated in HIF1A polyclonal antibody (Product # PA1-16627). This experiment was performed under reducing conditions using the 12-230 kDa separation system.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of HIF1A in cobalt chloride treated/untreated COS-7 nuclear extracts. Sample was incubated in HIF1A polyclonal antibody (Product # PA1-16627).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of HIF1A in 0.5 mg/mL Hypoxic HeLa lysate. Samples were incubated in HIF1A polyclonal antibody (Product # PA1-16627). This experiment was performed under reducing conditions using the 12-230 kDa separation system.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of HIF1A was performed using 70% confluent log phase HeLa cells treated with 150µM of CoCl2 for 48 hours. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with HIF1A Rabbit Polyclonal Antibody(Product # PA1-16627) at 5 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear localization. Panel e shows untreated cells with cytoplasmic signal. 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 formalin-fixed paraffin-embedded tissue section of human endometrium carcinoma AN3CA cell line based xenograft. Samples were incubated in HIF1A polyclonal antibody (Product # PA1-16627) using a dilution of 1:300. The signal was developed using HRP-labelled secondary antibody and DAB reagent, and 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).