Rabbit anti grp78

Manufactured by Proteintech

Sourced in United States

Rabbit anti-GRP78 is a primary antibody that recognizes the 78 kDa glucose-regulated protein (GRP78), also known as BiP. GRP78 is a molecular chaperone that plays a crucial role in the endoplasmic reticulum stress response. This antibody can be used to detect and study the expression and localization of GRP78 in various biological samples.

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4 protocols using rabbit anti grp78

Quantifying Immune Signaling in Brain Tissue

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Hippocampi were homogenized on ice in 2% SDS, 95 mM NaCl, 25 mM Tris, pH 7.4, 10 mM EDTA, and protease inhibitor mixture (Roche, Switzerland). After centrifugation, extracts were used for Western blot. Extracts from brains, as well as cultured BV2 and N2a cells, were run on reducing SDS/PAGE gels, transferred to PVDF membrane (EMD Millipore), incubated overnight at 4 °C in primary antibody [rabbit anti-TLR4 (Wanleibio, China); rabbit anti-MyD88 (Abcam); rabbit anti-phospho-NF-κB p65 (Ser536) (Cell Signaling Technology, USA); rabbit anti-NF-κB p65 Rabbit mAb (Cell Signaling Technology); rabbit anti-GRP78 (ProteinTech)], and then incubated with an appropriate HRP-conjugated secondary antibody. Blots were developed with high-intensity chemiluminescence Western blotting substrate (Tanon, China) and captured by Tannon 5200Multi Imager (Tannon). Protein levels were quantified using densitometry with ImageJ software. Western blot results for brain protein extracts were normalized to actin.

Xu J., Yang C., Zeng S., Wang X., Yang P, & Qin L. (2023). Disturbance of neuron–microglia crosstalk mediated by GRP78 in Neuropsychiatric systemic lupus erythematosus mice. Journal of Neuroinflammation, 20, 150.

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Immunofluorescence Staining of Cell Junctions

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For immunofluorescence staining, cells or spinal cord sections were fixed in 4% PFA for 15 min and subsequently permeabilized in 0.2% Triton X-100/PBS for 20 min. The samples were treated with 5% bovine serum albumin (BSA) for 1 h at room temperature to block nonspecific binding. The samples were incubated overnight at 4 °C with primary antibody specific for rabbit anti-ZO1 (1:50, Invitrogen), rabbit anti-claudin-5 (1:100; Invitrogen), rabbit anti-occludin (1:50; Abcam), mouse anti-CD31 (1:200, Invitrogen), mouse anti-NeuN (1:500, Abcam), and rabbit anti-GRP78 (1:200, Proteintech). Samples were washed thrice with PBS and incubated with Alexa 594- or Alexa 488-conjugated secondary antibodies (1:200, Jackson ImmunoResearch, USA) for 1 h in the dark. Finally, nuclei were labeled with DAPI and images were obtained under similar exposure time and conditions.

Lu X., Lv C., Zhao Y., Wang Y., Li Y., Ji C., Wang Z., Ye W., Yu S., Bai J, & Cai W. (2022). TSG-6 released from adipose stem cells-derived small extracellular vesicle protects against spinal cord ischemia reperfusion injury by inhibiting endoplasmic reticulum stress. Stem Cell Research & Therapy, 13, 291.

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Antibody and Chemical Reagents for PRRSV Study

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Primary antibodies were mouse anti-PRRSV N protein (produced in our laboratory), mouse anti-β-actin (Proteintech, #A5316), mouse anti-Flag (Proteintech, #80010-1-RR), rabbit anti-LC3 (Proteintech, #14600-1-AP), rabbit anti-ATG7 (Cell Signaling Technology, #2631), rabbit anti-CaMKII (Abcam, #ab168818), rabbit anti-p-AMPK (Thr172) (Cell Signaling Technology, #2535), rabbit anti-AMPK (Cell Signaling Technology, #2532), rabbit anti-p-mTOR(Ser2448) (Cell Signaling Technology, #5536), rabbit anti-mTOR (Cell Signaling Technology, #2983), rabbit anti-STIM1 (Cell Signaling Technology, #4916), mouse anti-Orai1 (Proteintech, #66223–1), rabbit anti-GRP78 (Proteintech, #11587-1-AP) and rabbit anti-Calnexin (Abcam, #ab92573). HRP-labeled rabbit or mouse secondary antibodies were purchased from Beyotime (China).
Chemicals used included 2-Aminoethyl diphenylborinate (2-APB) (Selleck, #S6657), 3-MA (Selleck, #S2767), 4-Phenylbutyric acid (4-PBA) (Selleck, #S3592), BAPTA-AM (Selleck, #S7534), Dantrolene sodium (Selleck, #S5478), Dorsomorphin (Compound C) 2HCL (Selleck, #S7306), KN-93 (Selleck, #S7423), ML-9 HCL (Selleck, #S6847), Procaine (Selleck, #S4668), Rapamycin (MedChemExpress, #HY-10219), Tetracaine-HCL (Selleck, #S2573), Thapsigargin (Selleck, #S7895), Tunicamycin (Beyotime, #SC0393).

Diao F., Jiang C., Sun Y., Gao Y., Bai J., Nauwynck H., Wang X., Yang Y., Jiang P, & Liu X. (2023). Porcine reproductive and respiratory syndrome virus infection triggers autophagy via ER stress-induced calcium signaling to facilitate virus replication. PLOS Pathogens, 19(3), e1011295.

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BMEC Proteome Response to NEFA Exposure

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The BMECs were grown in a 6-well plate and treated with various NEFA concentrations (0, 0.3, 0.6, and 0.9 mM) for 6 h. Whole BMECs protein lysates were centrifuged at 15,000 rpm for 15 min at 4 °C, and supernatants were quantified for total protein using a BCA protein assay kit (Beyotime, Beijing, China), according to the manufacturer’s instructions. Proteins were separated by SDS-PAGE and transferred to a PVDF membrane. The membranes were probed with the following primary antibodies: rabbit anti-GRP78, Bcl2, Caspase3, CHOP, IRE1 (1:1000, Proteintech, Chicago, IL, USA), rabbit anti-HSP70, PERK, phosphor IRE1, XBP1, ATF6, p38 MAPK, phosphor-p38 MAPK, ERK, phosphor-ERK, JNK and phosphor-JNK (1:1000, ABclonal, Boston, MA, USA), rabbit anti-ATF4, phosphor PERK (1:500, Bioss, Beijing, China), rabbit anti-Bax, and Tubulin (1:1000, Bioworld, Louis Park, MN, USA). The blots were incubated with HRP-conjugated secondary antibodies, and the signals were detected by enhanced chemiluminescence (ECL) Western blot detection reagents (Pierce, Rockford, IL, USA). Immunoblots were scanned and densitometry was performed using ImageJ software (v 1.48, National Institutes of Health, Bethesda, MD, USA).

Yan Y., Huang J., Huan C., Li L, & Li C. (2022). Non-Esterified Fatty Acid Induces ER Stress-Mediated Apoptosis via ROS/MAPK Signaling Pathway in Bovine Mammary Epithelial Cells. Metabolites, 12(9), 803.

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