Anti-PHGDH/Malate dehydrogenase antibody [4A3-1D6]

• Flow Cyt

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Flow Cytometry - Anti-PHGDH/Malate dehydrogenase antibody [4A3-1D6] (AB57030)

Overlay histogram showing HeLa cells stained with ab57030 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab57030, 1μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was DyLight^® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.

This image was generated using the ascites version of the product.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHGDH/Malate dehydrogenase antibody [4A3-1D6] (AB57030)

Immunohistochemical analysis of formalin-fixed paraffin-embedded human lymph node tissue labeling PHGDH with an antibody concentration of 3 µg/ml.

Positive staining on human lymph node tissue.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHGDH/Malate dehydrogenase antibody [4A3-1D6] (AB57030)

Immunohistochemical analysis of formalin-fixed paraffin-embedded human prostate tissue labeling PHGDH with an antibody concentration of 3 µg/mL.

Positive staining on human prostate tissue.

• IP

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Immunoprecipitation - Anti-PHGDH/Malate dehydrogenase antibody [4A3-1D6] (AB57030)

PHGDH/Malate dehydrogenase was immunoprecipitated using 0.5mg Jurkat whole cell extract, 10μg of Mouse monoclonal to PHGDH/Malate dehydrogenase and 50μl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Jurkat whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40μl SDS loading buffer and incubated for 10min at 70^oC; 10μl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab57030.
Secondary : Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution.
Band : 57kDa; PHGDH/Malate dehydrogenase.

This image was generated using the ascites version of the product.

All lanes:

Immunoprecipitation - Anti-PHGDH/Malate dehydrogenase antibody [4A3-1D6] (ab57030)

Predicted band size: 57 kDa

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• WB

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Western blot - Anti-PHGDH/Malate dehydrogenase antibody [4A3-1D6] (AB57030)

PHGDH/Malate dehydrogenase antibody (ab57030) at 1μg/ml + Jurkat cell lysate at 25μg/lane.

This image was generated using the ascites version of the product.

All lanes:

Western blot - Anti-PHGDH/Malate dehydrogenase antibody [4A3-1D6] (ab57030)

Predicted band size: 57 kDa

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• WB

CiteAb

Western blot - Anti-PHGDH/Malate dehydrogenase antibody [4A3-1D6] (AB57030)

PHGDH/Malate dehydrogenase western blot using anti-PHGDH/Malate dehydrogenase antibody [4A3-1D6] ab57030. Publication image and figure legend from Kitazawa, S., Ebara, S., et al., 2017, Oncotarget, PubMed 28423651.

ab57030 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab57030 please see the product overview.

Metabolic switch from aerobic metabolism to glycolysis in SDHB-deficient cells(A) Relative values of OCR and ECAR of HCT116 SDHB knockout cells (#7 and #23) were obtained by setting the OCR and ECAR values of the control cells as 1. The OCR and ECAR were analyzed using an XF96 extracellular flux analyzer (n = 12). Data are given as means ± SD. (B) Metabolite concentrations of pyruvate and lactate were measured using CE-MS 48 and 96 h after cell seeding in HCT116 SDHB knockout cells (#7 and #23) and control cells (n = 3). Data are given as means ± SD. *p < 0.05, **p < 0.01, ***p < 0.001 by the Bonferroni's corrected t-test. (C) Metabolites and enzymes in the glycolysis pathway and its derived pathways. Metabolite abundance 48 h after cell seeding was analyzed using CE-MS (n = 3). Ordinate values were obtained by setting the control group value 48 h after cell seeding as 100%. Data are given as means ± SD. ND : not detected. ND : the metabolite concentration was below the detection limit of the analysis. (D) Expression levels of SDHB, PHGDH, PSAT1, PSPH, and β-actin were determined by western blot 64 h after cell seeding.

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