rabbit anti gapdh antigen affinity purified pab  (Boster Bio)

Journal: Open Medicine

Article Title: EphA3 targeted by miR-3666 contributes to melanoma malignancy via activating ERK1/2 and p38 MAPK pathways

doi: 10.1515/med-2022-0597

Figure Lengend Snippet: Real-time PCR primer synthesis list

Article Snippet: Proteins (20 μg per lane) were resolved in 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis and detected on poly(vinylidene fluoride) membranes after which was blocked by 5% skim milk powder at room temperature for 2 h. Protein detection on membranes was detected with anti-p38MAPK (FNab06077, 1:1,000, FineTest, Biological Technology Co., Ltd, China), anti-p-p38MAPK (44-684G, 1:1,000, ThermoFisher), anti-p-ERK1/2 (44-680G, 1:1,000, ThermoFisher), anti-ERK1/2 (13-6200, 1:1,000,ThermoFisher), anti-EphA3 (310438-T08, 1:1,000, SinoBiological, China), and anti-GAPDH (100242-T08, 1:1,000, SinoBiological) at 4°C for 24 h. Next, horseradish peroxidase-conjugated secondary antibodies (A32731, 1:1,000, ThermoFisher) were added and incubated with these membranes at room temperature for 1 h. Immune blots were developed with enhanced chemiluminescence (GE Healthcare, USA) and counted quantitatively by ImageJ.

Techniques: Real-time Polymerase Chain Reaction

Journal: Oncology Letters

Article Title: Serum anti-GAPDH autoantibody levels reflect the severity of cervical lesions: A potential serum biomarker for cervical cancer screening

doi: 10.3892/ol.2019.10326

Figure Lengend Snippet: Reactivity of sera from healthy females against HeLa cell cytosolic proteins using western blotting. The arrow indicates a major HeLa cell cytosolic protein detected by the sera from healthy females. The major protein was identified as GAPDH isoform-1 using nano ACQUITY ultra performance liquid chromatography coupled directly to a linear trap quadrupole-orbitrap-mass spectrometer. β-actin polyclonal antibody was used as a positive internal loading control in western blotting.

Article Snippet: Mixtures of sera from the group of control females (n=20) and rabbit anti-human GAPDH polyclonal antibody (cat. no.10094-T52, Sino Biological, Beijing, China) were diluted 1:200 in TBS containing 0.1% Tween-20 (TBST) and 2.5% skimmed milk, and 1:1000 in TBST containing 0.5% skimmed milk, respectively, and reacted with the membranes for 2 h at room temperature.

Techniques: Western Blot, Liquid Chromatography, Mass Spectrometry

Journal: Oncology Letters

Article Title: Serum anti-GAPDH autoantibody levels reflect the severity of cervical lesions: A potential serum biomarker for cervical cancer screening

doi: 10.3892/ol.2019.10326

Figure Lengend Snippet: Purification of HeLa-GAPDH from HeLa cell cytosolic proteins by cation-exchange chromatography. (A) HeLa cell cytosolic proteins were separated by cation-exchange chromartography. The proteins were separated by SDS-PAGE and visualized with silver staining. (B) Western blot analysis of proteins from cation-exchange chromatography using healthy donor sera. (C) Western blot analysis where HeLa-GAPDH was detected in fractions from cation-exchange chromatography using anti-human GAPDH polyclonal antibody. HeLa-GAPDH is indicated by arrowheads. M, protein markers; LS, loading sample; FT, flow-through; CW, column wash.

Article Snippet: Mixtures of sera from the group of control females (n=20) and rabbit anti-human GAPDH polyclonal antibody (cat. no.10094-T52, Sino Biological, Beijing, China) were diluted 1:200 in TBS containing 0.1% Tween-20 (TBST) and 2.5% skimmed milk, and 1:1000 in TBST containing 0.5% skimmed milk, respectively, and reacted with the membranes for 2 h at room temperature.

Techniques: Purification, Chromatography, SDS Page, Silver Staining, Western Blot