PA5-53023

antibody from Invitrogen Antibodies

Targeting: VAPB ALS8, VAP-B, VAP-C

Western blot

Immunoprecipitation

Immunohistochemistry

Antibody data

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Validation data

Product number: PA5-53023 - Provider product page
Provider: Invitrogen Antibodies
Product name: VAPB Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Recombinant full-length protein
Description: Immunogen sequence: VWKEAKPEDL MDSKLRCVFE LPAENDKPHD VEINKIISTT ASKTETPIVS KSLSSSLDDT EVKKVMEECK RLQGEVQRLR EENKQFKEED GLRMRKTVQS NSPISALAPT GK Highest antigen sequence identity to the following orthologs: Mouse - 82%, Rat - 79%.
Reactivity: Human, Mouse, Rat
Host: Rabbit
Isotype: IgG
Vial size: 100 µL
Concentration: 0.3 mg/mL
Storage: Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

VAPB protein structure - PA5-53023 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
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)
Main image
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)
Main image
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)
Main image
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