polyclonal rabbit anti human nterminal vapb antibody  (Proteintech)

Journal: European journal of neurology

Article Title: Expression of vesicle-associated membrane-protein-associated protein B cleavage products in peripheral blood leukocytes and cerebrospinal fluid of patients with sporadic amyotrophic lateral sclerosis.

doi: 10.1111/ene.12334

Figure Lengend Snippet: Figure 1 Schematic representation of structural domains of human VAPB protein. Positions of VAPB mutations (p.P56S, p.T46I and p.V234I) and immunogen peptide (31–180 aa) of the wild-type VAPB used to generate the anti-human VAPB antibody (Proteintech Group Inc.) are shown.

Article Snippet: Membranes blocked with TBS buffer plus 0.1% Tween-20 and 5% non-fat milk for 1 h were incubated with a polyclonal rabbit anti-human (Nterminal) VAPB antibody (1:400, Proteintech Group Inc., Chicago, IL, USA) overnight at 4°C.

Techniques:

Journal: European journal of neurology

Article Title: Expression of vesicle-associated membrane-protein-associated protein B cleavage products in peripheral blood leukocytes and cerebrospinal fluid of patients with sporadic amyotrophic lateral sclerosis.

doi: 10.1111/ene.12334

Figure Lengend Snippet: Figure 2 Expression of VAPB and VAPA proteins in PBL and CSF from healthy (HC) and neurological (NC) controls. (a), (b), (d) Representative immunoblots showing the expression pattern of VAPB (a), (b) and VAPA (d) identified in PBL and CSF. (c) Peptide competition experiments to verify specificity of the anti- VAPB antibody. Western blots were carried out with (a) 100 lg or (b)–(d) 65 lg proteins of extracts. Differences in the molecu- lar sizes of VAPB and VAPA bands are reported in leucocytes. VAPB and VAPA exhibit two close bands that approximate the molecular size of 17 kDa. In CSF, only the 14 kDa band of VAPB was identified in most samples examined, and no VAPA product was detected; in rare instances, a 24 kDa band of VAPB (asterisk) was additionally detected.

Article Snippet: Membranes blocked with TBS buffer plus 0.1% Tween-20 and 5% non-fat milk for 1 h were incubated with a polyclonal rabbit anti-human (Nterminal) VAPB antibody (1:400, Proteintech Group Inc., Chicago, IL, USA) overnight at 4°C.

Techniques: Expressing, Western Blot

Journal: European journal of neurology

Article Title: Expression of vesicle-associated membrane-protein-associated protein B cleavage products in peripheral blood leukocytes and cerebrospinal fluid of patients with sporadic amyotrophic lateral sclerosis.

doi: 10.1111/ene.12334

Figure Lengend Snippet: Figure 3 Scatter plots of the expression levels of VAPB MSP fragments in PBL of SALS patients and controls. Levels of the 17 kDa (a) or 14 kDa (b) fragments are expressed as the ratio between their intensities and those of b-actin immunoreactive bands. No significant differences for the two fragments were found amongst HC, NC and SALS.

Article Snippet: Membranes blocked with TBS buffer plus 0.1% Tween-20 and 5% non-fat milk for 1 h were incubated with a polyclonal rabbit anti-human (Nterminal) VAPB antibody (1:400, Proteintech Group Inc., Chicago, IL, USA) overnight at 4°C.

Techniques: Expressing

Journal: European journal of neurology

Article Title: Expression of vesicle-associated membrane-protein-associated protein B cleavage products in peripheral blood leukocytes and cerebrospinal fluid of patients with sporadic amyotrophic lateral sclerosis.

doi: 10.1111/ene.12334

Figure Lengend Snippet: Figure 4 Analyses of the VAPB MSP fragment in CSF from neurological controls (NC) and SALS patients with bulbar (ALS-B) and spinal onset (ALS-S). (a) Representative immunoblots of the CSF 14 kDa fragment from 15 subjects analysed of each group; albumin immunoblotting was used to control protein loading. (b), (c) Scatter plots of the expression levels of the CSF VAPB MSP fragment detected in the samples examined (NC, n = 33; SALS, n = 46; ALS-S, n = 22; ALS-B, n = 24). Western blot analyses of VAPB MSP were performed in triplicate using the same amount of proteins (65 lg) for each CSF sample and by reprobing membranes with albu- min antibody. Levels of the 14 kDa fragment result from the ratio of the fragment over albumin density in each sample on western blotting. (a) *Mann–Withney U test (P < 0.0001, NC vs. SALS); (b) *Kruskal–Wallis test with Dunn’s multiple comparison test (P < 0.0001, NC vs. ALS-B).

Article Snippet: Membranes blocked with TBS buffer plus 0.1% Tween-20 and 5% non-fat milk for 1 h were incubated with a polyclonal rabbit anti-human (Nterminal) VAPB antibody (1:400, Proteintech Group Inc., Chicago, IL, USA) overnight at 4°C.

Techniques: Western Blot, Control, Expressing, Comparison