- MA5-15776 - Invitrogen Antibodies ORAI1 antibody

Submitted references Expression of Orai1 and STIM1 in human oral squamous cell carcinogenesis.

Wang YY, Wang WC, Su CW, Hsu CW, Yuan SS, Chen YK
Journal of dental sciences 2022 Jan;17(1):78-88

Differential activation of Ca(2+) influx channels modulate stem cell potency, their proliferation/viability and tissue regeneration.

Ahamad N, Sun Y, Nascimento Da Conceicao V, Xavier Paul Ezhilan CRD, Natarajan M, Singh BB
NPJ Regenerative medicine 2021 Oct 20;6(1):67

Elevated serotonin coordinates mammary metabolism in dairy cows.

Connelly MK, Weaver SR, Kuehnl JM, Fricke HP, Klister M, Hernandez L
Physiological reports 2021 Apr;9(7):e14798

Cannabinoid signalling inhibits sarcoplasmic Ca(2+) release and regulates excitation-contraction coupling in mammalian skeletal muscle.

Oláh T, Bodnár D, Tóth A, Vincze J, Fodor J, Reischl B, Kovács A, Ruzsnavszky O, Dienes B, Szentesi P, Friedrich O, Csernoch L
The Journal of physiology 2016 Dec 15;594(24):7381-7398

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of human ovary tissue lysate using a ORAI1 monoclonal antibody (Product # MA5-15776) at (A) 1 and (B) 2 µg/mL.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot analysis of ORAI1 in human ovary tissue lysate with ORAI1 Monoclonal Antibody (3F6H5) (Product # MA5-15776) at (A) 1 and (B) 2 µg/mL.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of ORAI1 was achieved by transfecting Caco-2 with ORAI1 specific siRNAs (Silencer® select Product # s228396, s39559). Western blot analysis (Fig. a) was performed using whole cell extracts from the ORAI1 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with ORAI1 Monoclonal Antibody (3F6H5) (Product # MA5-15776, 2µg/ml) and Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/ml 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to ORAI1.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis was performed on whole cell extracts (30 µg lysate) of HeLa (Lane 1), MCF7 (Lane 2), DU 145 (Lane 3), NIH/3T3 (Lane 4), C2C12 (Lane 5) and SK-OV-3 (Lane 6). The blot was probed with Anti-ORAI1 Monoclonal Antibody (Product # MA5-15776, 2 µg/ml) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/ml, 1:4000 dilution). A 50 kDa band corresponding to ORAI1 was observed across the cell lines tested.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescent analysis of human spleen cells using a ORAI1 monoclonal antibody (Product # MA5-15776) at a 20 µg/mL dilution.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence of ORAI1 in human spleen tissue with ORAI1 Monoclonal Antibody (3F6H5) (Product # MA5-15776) at 20 µg/mL.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence of ORAI1 in human spleen tissue with ORAI1 Monoclonal Antibody (3F6H5) (Product # MA5-15776) at 20 µg/mL. Red: ORAI1 Antibody [3F6H5] (PM-5205) Blue: DAPI staining

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry of ORAI1 in human spleen tissue with ORAI1 Monoclonal Antibody (3F6H5) (Product # MA5-15776) at 2.5 µg/mL.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry of ORAI1 in human spleen tissue with ORAI1 Monoclonal Antibody (3F6H5) (Product # MA5-15776) at 5 µg/mL.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 4 MSCs cell viability and cell proliferation after treatment of calcium channel blockers. a Western blot showing expression of calcium channel proteins in MSCs. b Application of 2 uM thapsigargin (Tg) in bath solution induced an inward Ca 2+ current which is shown at -80 mV in MSC cells. c Respective IV curves of Ca 2+ currents and its quantitation (5-8 recordings) of current intensity at -80 mV are shown in d . e IV curves of Ca 2+ currents in control non-targeted siRNA (NTsiRNA) and TRPC1siRNA-treated cells. Quantitation (7-9 recordings) of current intensity in individual conditions at -80 mV are shown in f . * Indicate significance ( p < 0.05). g 2APB-induced Ca 2+ currents at -100 and +100 mV. h Respective IV curves and its quantitation (6-8 recordings) of 2APB-induced current intensity in control and SKF-treated cells. i IV curves of 2APB-induced Ca 2+ currents in control non-targeted scrambled siRNA (NTsiRNA) and Orai3siRNA-treated cells. Quantitation (5-6 recordings) of current intensity in individual conditions at -80 mV are shown in j . * Indicate significance ( p < 0.05). k , l MSCs were silenced with individual siRNAs (STIM1, TRPC1, Orai1, and Orai3), and cell viability and proliferation were studied. NS indicates non-significant, whereas, *Significance p

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: FIGURE 4 Milk calcium concentrations (a) ( n = 6), mammary gland mRNA expression of PMCA2 (b) (5-HTP n = 6, n = 6, n = 5; Control n = 6, n = 6, n = 6), PTHLH (c) (5-HTP n = 6, n = 5, n = 6; Control n = 4, n = 4, n = 5), CASR (d) (5-HTP n = 6, n = 5, n = 5; Control n = 3, n = 4, n = 3), ORAI1 (e) (5-HTP n = 6, n = 6, n = 6; Control n = 5, n = 6, n = 6), quantification and western blot of mammary gland ORAI1 (f-g) (5-HTP n = 4, n = 4, n = 3; Control n = 4, n = 4, n = 3), mammary mRNA expression of NCX1 (h) (5-HTP n = 6, n = 6, n = 6; Control n = 6, n = 5, n = 5), SPCA1 (i) (5-HTP n = 6, n = 6, n = 6; Control n = 5, n = 6, n = 5), and SERCA2 (j) (5-HTP n = 6, n = 6, n = 6; Control n = 5, n = 4, n = 3) in multiparous Holstein dairy cows receiving 1 L of 1.5 mg/kg 5-HTP dissolved in saline or 1 L of saline solution (Saline = Control) for three consecutive days. Statistics were performed using PROC MIXED procedure with repeated measures in SAS. Transformed data presented as mean +- SEM. Normally distributed data presented as LSMEANS +- SEM. Milk calcium: treatment ( p = 0.02). ORAI1 mRNA: treatment ( p = 0.09) and treatment*time ( p = 0.06). PTHLH mRNA: time ( p = 0.04) and treatment*time ( p = 0.03). Asterisks denote statistical significance between groups (** p < 0.01, * p < 0.05 and #0.10 < p > 0.05)

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 1 Immunohistochemistry for Orai1 (A) A representative strong immunohistochemical staining of Orai1 protein in human oral squamous cell carcinoma (x 100) (B) A representative weak staining of Orai1 protein in human normal oral mucosa (x 100; x 400 for inset) (C) A representative stronger immunohistochemical staining of Orai1 protein for a human oral potentially malignant disorder with moderate to severe oral epithelial dysplasia (x 100) (D) A representative weaker staining of Orai1 protein for a human oral potentially malignant disorder with mild oral epithelial dysplasia (x 100). Figure 1

MA5-15776

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