PA5-27359

antibody from Invitrogen Antibodies

Targeting: G6PD G6PD1

Western blot (Data presented on provider website)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-27359

Provider

Invitrogen Antibodies

Product name

G6PD Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Recombinant protein fragment

Description

Recommended positive controls: 293T, A431, H1299, HeLaS3, HepG2, Raji, Neuro 2A, C8D30, NIH-3T3, Raw264.7, C2C12, PC-12, Rat2. Predicted reactivity: Rat (93%), Xenopus laevis (82%), Rabbit (95%), Sheep (92%), Rhesus Monkey (99%), Bovine (95%). Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.

Reactivity

Human, Mouse, Rat

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Concentration

0.81 mg/mL

Storage

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

References

Submitted references

Breast cancer stem cells rely on fermentative glycolysis and are sensitive to 2-deoxyglucose treatment.

Ciavardelli D, Rossi C, Barcaroli D, Volpe S, Consalvo A, Zucchelli M, De Cola A, Scavo E, Carollo R, D'Agostino D, Forlì F, D'Aguanno S, Todaro M, Stassi G, Di Ilio C, De Laurenzi V, Urbani A
Cell death & disease 2014 Jul 17;5(7):e1336

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p63 isoforms regulate metabolism of cancer stem cells.

D'Aguanno S, Barcaroli D, Rossi C, Zucchelli M, Ciavardelli D, Cortese C, De Cola A, Volpe S, D'Agostino D, Todaro M, Stassi G, Di Ilio C, Urbani A, De Laurenzi V
Journal of proteome research 2014 Apr 4;13(4):2120-36

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of G6PD was performed using HeLa cells. 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 1 hour at room temperature. The cells were labeled with G6PD Goat Polyclonal Antibody (Product # PA5-27359) at 1:100 dilution in 0.1% BSA and incubated overnight at 4 degree and then labeled with Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Recombinant 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 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 composite image showing Nuclear and Cytoplasmic localization of G6PD. Panel e 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 paraffin-embedded U87 xenograft, using G6PD (Product # PA5-27359) antibody at 1:300 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 4 The expression levels and activities of glycolytic enzymes are increased in BCSCs and correlate with altered energy and redox status of BCSCs. ( a ) WB showing PKM2, LDH-A, and G6PDH expression in BCSCs compared with SDACs. beta -Actin is used as a protein-loading control. A representative WB of three independent experiments is shown. ( b ) PK and ( c ) LDH, and enzyme activities evaluated in spheres (BCSCs) as compared with differentiated cells (SDACs). Bar graphs show the mean activity ( n =6) expressed as mu mol/mg Protein /min, and bars represent the S.E.M. ( d ) WB showing the expression of PKM2 and LDH-A in cells sorted for CD24 expression. beta -Actin is used as a loading control. ( e ) PK and ( f ) LDH activity in CD24 -/low cells compared with CD24 + BCSCs. Bar graphs show the mean activity ( n =6) expressed as mu mol/mg Protein /min, and bars represent the S.E.M. ( g ) G6PDH enzyme activity evaluated in spheres (BCSCs) as compared with differentiated cells (SDACs). Bar graphs show the mean activity ( n =6) expressed as mu mol/mg Protein /min, and bars represent the S.E.M. ( h ) NADP + /NADPH, ( i ) NAD + /NADH, and ( j ) ADP/ATP ratios in BCSCs compared with SDACs. Treatment with rotenone does not affect ATP production in BCSCs, whereas strongly increases the ADP/ATP ratio and inhibits mitochondrial ATP production in SDACs. No changes in the ADP/ATP ratio are observed in BCSCs and SDACs after addition of dimethylsulphoxide (DMSO) alone used as a vehicle con