Rabbit anti stim1

Manufactured by Cell Signaling Technology

Sourced in United States

Rabbit anti-STIM1 is a polyclonal antibody that recognizes the STIM1 (Stromal Interaction Molecule 1) protein. STIM1 is a calcium sensor that plays a crucial role in regulating calcium homeostasis and signaling in cells.

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STIM1 (20 citations) Anti-STIM1 (10 citations) Rabbit monoclonal anti-human STIM1 antibody (2 citations)

5 protocols using rabbit anti stim1

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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Antibody Immunoprecipitation and Immunoblotting Protocol

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The following antibodies were used in for protein immunoprecipitation or immunoblotting: mouse anti-Flag M2 (Sigma), rabbit anti-Flag (Sigma), anti-GAPDH (Calbiochem), rabbit anti-Stim1 (Cell Signaling), anti-Bip/GRP78 (Abcam), anti-PARP (cleaved) (Cell Signaling), anti-Calsequestrin 2 (Thermo), anti-Fam20C (Tagliabracci et al., 2014 (link)), and anti-His (Invitrogen).

Pollak A.J., Liu C., Gudlur A., Mayfield J.E., Dalton N.D., Gu Y., Chen J., Heller Brown J., Hogan P.G., Wiley S.E., Peterson K.L, & Dixon J.E. (2018). A secretory pathway kinase regulates sarcoplasmic reticulum Ca2+ homeostasis and protects against heart failure. eLife, 7, e41378.

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Antibody Validation for Western Blot and IF

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The following antibodies were used for western blotting (WB) and immunofluorescence (IF): mouse anti-beta-actin (Santa Cruz, catalog number sc-47778; WB 1:2000), mouse anti-HA (ENZO, catalog number ENZ-ABS120-0200; WB 1:1000), mouse anti-transferrin receptor (Invitrogen, catalog number 13–6800; WB 1:1000), rabbit anti-Stim1 (Cell Signaling Technology, catalog number 5668S (D88E10); WB 1:2000), rabbit anti-Orai1 (Sigma Aldrich, catalog number O8264; WB 1:2500), goat anti-myc tag (Abcam, catalog number ab9132; IF 1:2000), rabbit anti-RHBDL2 (Proteintech, catalog number 12467-1-AP; WB 1:250 – only detected RHBDL2 in HaCaT lysates), rabbit anti-V5 tag (Cell Signaling Technology, catalog number 13202S; WB and IF 1:2000). Corresponding species-specific HRP or fluorescently coupled secondary antibodies were used from Santa Cruz and Cell Signaling (WB) or Invitrogen (IF).

Grieve A.G., Yeh Y.C., Chang Y.F., Huang H.Y., Zarcone L., Breuning J., Johnson N., Stříšovský K., Brown M.H., Parekh A.B, & Freeman M. (2021). Conformational surveillance of Orai1 by a rhomboid intramembrane protease prevents inappropriate CRAC channel activation. Molecular Cell, 81(23), 4784-4798.e7.

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Immunostaining of Brain Tissues and Cells

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Brain tissues were fixed in 4 % paraformaldehyde for 24 h at room temperature (RT) and dehydrated in phosphate-buffered saline (PBS) containing 30 % sucrose for 48 h at 4 °C and then coronally sectioned into 30 μm thick slices with a cryostat. Cells were fixed in 4 % paraformaldehyde for 30 min at RT. 30 μm-thick frozen coronal sections or cultured cells were first permeabilized in 0.3 % Triton X-100 for 20 min, incubated with 10 % goat serum for 1 h at RT. The sections or cells were then incubated at 4 °C overnight with the primary antibodies, including mouse anti-shank3 (Neuromab, Cat# 75344, 1:200), rabbit anti-STIM1 (1:100, Cell Signaling Technology, Cat# 5668), rabbit anti-NeuN (1:100, Abcam, Cat# ab177487). After rinsing three times with PBS, sections or cells were incubated with secondary antibodies for 2 h at RT in the dark. Slices were covered with a mounting medium containing DAPI to counterstain nuclei and imaged using a laser scan confocal microscope (FV1000, Olympus, Japan) with the same exposure settings for each group.

Zhang H., Feng Y., Si Y., Lu C., Wang J., Wang S., Li L., Xie W., Yue Z., Yong J., Dai S., Zhang L, & Li X. (2023). Shank3 ameliorates neuronal injury after cerebral ischemia/reperfusion via inhibiting oxidative stress and inflammation. Redox Biology, 69, 102983.

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Western Blot Analysis of Protein Expression

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Cells were lysed in a protein extraction solution (Intron, Seongnam, South Korea). After centrifugation, the samples were boiled at 95 °C for 10 min, and 50 µg protein of each lysate was subjected to electrophoresis on 10% sodium dodecyl sulfate polyacrylamide gels. All samples were electroblotted on polyvinylidene fluoride membranes (Merck Millipore, Burlington, MA, USA) and, after blocking, the blots were incubated with appropriate rabbit anti-THBS4 (Santa Cruz Biotechnology, sc-390734), rabbit anti-PDGFRβ (Abcam, Cambridge, MA, USA, ab32570), mouse anti-phosphorylated PDGFRβ (Santa Cruz Biotechnology, sc-365464), rabbit anti-PDGF-D (Abcam, Cambridge, MA, USA, ab234666), mouse anti-IP3R (Santa Cruz Biotechnology, sc-271197), and rabbit anti-STIM1 (Cell Signaling Technology, Danvers, MA, USA, 5668S) antibodies in TBS-T (TBS-0.05% Tween 20) for 90 min, which was followed by washing three times with TBS-T for 15 min each, and incubation with horseradish peroxidase-conjugated anti-mouse or rabbit immunoglobulin G antibodies for 1 h. After further washing, the blots were incubated for 3 min with Western blotting HRP-substrate (Merck Millipore), and chemiluminescence was detected after exposure of the filters to ECL-Western blot films for 10 s to 10 min. Original Western blots are shown on Figures S7–S14.

Kim M.S., Choi H.S., Wu M., Myung J., Kim E.J., Kim Y.S., Ro S., Ha S.E., Bartlett A., Wei L., Ryu H.S., Choi S.C., Park W.C., Kim K.Y, & Lee M.Y. (2020). Potential Role of PDGFRβ-Associated THBS4 in Colorectal Cancer Development. Cancers, 12(9), 2533.

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