- PA5-53023 - Invitrogen Antibodies VAPB antibody

Dorsch AD, Hölper JE, Franzke K, Zaeck LM, Mettenleiter TC, Klupp BG
Viruses 2021 Jun 10;13(6)

Bae H, Lee JY, Song J, Song G, Lim W
Journal of cellular physiology 2021 Feb;236(2):1025-1042

Bae H, Song G, Lim W
Pharmaceutics 2020 May 28;12(6)

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of VAPB in human cell lines Caco-2 and HEK293 using a VAPB Polyclonal Antibody (Product # PA5-53023). Corresponding VAPB RNA-seq data are presented for the same cell lines. Loading control: Anti-COX4I1.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of VAPB in mouse cell line NIH-3T3 and rat cell line NBT-II using a VAPB Polyclonal Antibody (Product # PA5-53023).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot of VAPB was performed by loading 50 µg of WT (lane 1) and VAPB CRISPR KO (lane 2) HeLa cell lysates in RIPA buffer onto a 5-16% gradient polyacrylamide gel. Proteins on the blots were visualized with Ponceau staining (below immunoblot). Proteins were transferred to nitrocellulose membrane and blocked in 5% milk for 1 hr. VAPB was detected at approximately 28 kDa using a VAPB polyclonal antibody (Product # PA5-53023) at a dilution of 1:250 in 5% BSA in TBS with 0.1% Tween 20 (TBST) overnight at 4°C. The peroxidase-conjugated secondary antibody (Product # 65-6120) was diluted to 0.2 µg/mL in TBST with 5% milk for 1 hr. Chemiluminescent detection was performed using Pierce ECL Western Blotting Substrate (Product # 32106). Data courtesy of YCharOS Inc., an open science company with the mission of characterizing commercially available antibodies using knockout validation.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescent staining of VAPB in human cell line U-2 OS shows positivity in endoplasmic reticulum. Samples were probed using a VAPB Polyclonal Antibody (Product # PA5-53023).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemical staining of VAPB in human cerebral cortex using a VAPB Polyclonal Antibody (Product # PA5-53023) shows strong cytoplasmic positivity in neuronal cells.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemical staining of VAPB in human small intestine using a VAPB Polyclonal Antibody (Product # PA5-53023) shows strong cytoplasmic positivity in glandular cells.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemical staining of VAPB in human kidney using a VAPB Polyclonal Antibody (Product # PA5-53023) shows moderate cytoplasmic positivity in cells in tubules.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemical staining of VAPB in human liver using a VAPB Polyclonal Antibody (Product # PA5-53023) shows moderate cytoplasmic positivity in hepatocytes.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunoprecipitation of VAPB was performed on HeLa cell lysates. Antibody-bead conjugates were prepared by adding 1 µg of VAPB polyclonal antibody (Product # PA5-53023) with 30 µL of protein A-Sepharose beads and rocked overnight at 4°C. 1 mg of lysate was incubated with an antibody-bead conjugate for 2 hours at 4°C. Following centrifugation and multiple washes, 10% starting material (SM), 10% unbound fraction (UB) and immunoprecipitated fraction (IP) were processed for immunoblot using a different VAPB antibody. Ponceau stained transfer of blot is shown. Data courtesy of YCharOS Inc., an open science company with the mission of characterizing commercially available antibodies using knockout validation.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 2 Immunoblots showing VAPA and VAPB in RK13, Vero and HeLa cells and the absence of VAPA and/or VAPB in the corresponding RK13 KO and DKO cell lines. Proteins in lysates of RK13, Vero and HeLa cells ( A ) or RK13 and the single as well as the double VAP KO cell lines ( B ) were separated on SDS-10 % polyacrylamide gels, and parallel blots were incubated with polyclonal sera against VAPA or VAPB. Masses of marker proteins are given on the left in kDa. As loading control, parallel ( A ) or the same blots were (re-)probed with anti-alpha-tubulin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 4 Increased expression of endoplasmic reticulum (ER) stress sensor proteins and ER-mitochondrial axis proteins due to stigmasterol treatment in ovarian cancer cells. ( A , B ) Immunoblots representing the expression of unfolded protein response (UPR) proteins following stigmasterol treatments (0, 5, 10, and 20 mug/mL). TUBA was used as a control and is shown at the bottom of the figure. ( C , D ) Immunoblots representing the expression of ER-mitochondrial axis proteins following stigmasterol treatments (0, 5, 10, and 20 mug/mL). TUBA was used as a control and is shown at the bottom of the figure. ( E , F ) Immunoblots representing the expression of autophagy proteins following stigmasterol treatment. TUBA was used as a control and is shown at the bottom of the figure.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: 8 Figure Osthole influenced intracellular mechanisms related to endoplasmic reticulum (ER)-mitochondrial functions. Expression of ER-mitochondrial function targets such as (a) ULK1, (b) GRP75, (c) MFN2, (d) P62, (e) FAM82A2, (f) VAPB, (g) VDAC, (h) IP3R1, and (i) IP3R2 was analyzed by western blot analysis in ES2 and OV90 cells treated with various doses of osthole (0, 5, 10, and 20 muM). Intensity of the target protein was detected and analyzed relative to total protein or alpha-tubulin protein. *** p < .001, ** p < .01, and * p < .05 indicate significant differences compared to vehicle-treated cells. IP3R1, inositol 1,4,5-trisphosphate (IP3) receptor type 1; IP3, inositol trisphosphate; MFN2, mitofusin-2; VAPB, vesicle-associated membrane protein-associated protein B/C; VDAC, voltage-dependent anion channel

PA5-53023