720178

antibody from Invitrogen Antibodies

Targeting: STMN2 SCG10, SCGN10

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

720178

Provider

Invitrogen Antibodies

Product name

STMN2 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Recombinant full-length protein

Reactivity

Human, Mouse, Rat

Host

Rabbit

Isotype

IgG

Vial size

100 µg

Concentration

0.5 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

References

Submitted references

Stathmin-2 Mediates Glucagon Secretion From Pancreatic α-Cells.

Asadi F, Dhanvantari S
Frontiers in endocrinology 2020;11:29

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The landscape of multiscale transcriptomic networks and key regulators in Parkinson's disease.

Wang Q, Zhang Y, Wang M, Song WM, Shen Q, McKenzie A, Choi I, Zhou X, Pan PY, Yue Z, Zhang B
Nature communications 2019 Nov 20;10(1):5234

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on whole cell extracts (30 µg lysate) of SH-SY5Y (Lane 1), IMR-32 (Lane 2), Rat Brain (Lane 3) and Mouse Brain (Lane4). The blots were probed with Anti- SCG10/Stathmin-2 Rabbit Polyclonal Antibody (Product # 720178, 0.5-1 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 21 kDa band corresponding to SCG10/Stathmin-2 was observed across cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris gel (Product # NP0321BOX), XCell SureLock Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence was performed on fixed and permeabilized SHSY-5Y cells for detection of Stathmin-2 using Anti-Stathmin-2 Rabbit Polyclonal Antibody (Product # 720178, 1 µg/mL), alpha-Tubulin was detected using Anti-alpha Tubulin Monoclonal Antibody (Product # 32-2500, 1 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000), Goat anti-Mouse IgG Secondary Antibody, Alexa Fluor®594conjugate (Product # A-11032, 1:400) respectively. Panel a) shows representative cells that were stained for detection and localization of Stathmin-2 protein (green), Panel b) is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938,). Panel c) represents cytoskeletal alpha-tubulin staining (red). Panel d) is a composite image of Panels a, b and c clearly demonstrating cytoplasmic localization of Stathmin-2. Panel e) represents control cells with no primary Antibody to assess background.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow Cytometry analysis of SCG10/Stathmin-2 was performed on SHSY5Y cells labeled with Anti-SCG10/Stathmin-2 Rabbit Polyclonal Antibody (Product# 720178, 2-4 ug/ 1M cells) or with Rabbit isotype control and detected with Goat anti-Rabbit IgG (H+L) Superclonalª Secondary Antibody, (Alexa Fluor¨ 488 conjugate, Product # A27034, 0.4 ug/ml, 1:2500) as represented by the red and pink histograms respectively. The purple histogram represents unstained control cells and the green histogram represents no-primary-Antibody control. A representative of 10,000 cells were acquired and analyzed for each sample using an Attune¨ Acoustic Focusing Cytometer (4468770).

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 4 Silencing Stathmin-2 increased glucagon secretion and overexpression of stathmin-2 suppressed glucagon secretion in alphaTC1-6 cells. Wild type (wt; control) and stathmin-2 depleted (Stmn2-KD) aTC1-6 cells were pre-incubated 2 h in serum-free medium and then incubated with or without KCl (55 mM) for 15 min. (A) Glucagon secretion is significantly stimulated by KCL in wt cells, while in Stmn-KD cells, basal glucagon secretion is increased and does not respond to KCl. * p < 0.01 compared to basal secretion in wt cells. (B) Stmn2 mRNA levels are decreased by about 70% after siRNA-mediated depletion in alphaTC1-6 cells. Values are means +- SEM ( n = 5), * p < 0.01. (C) Stathmin-2 protein levels are depleted after siRNA-mediated silencing of Stmn2. C (control); KD (gene silenced). Beta-actin was used as a loading control. (D) Proglucagon mRNA levels are not affected by siRNA-mediated depletion of stathmin-2. (E) Glucagon secretion is inhibited by overexpression of Stmn2. alphaTC1-6 cells were transfected with pcDNA3.1 (+) MAr-stmn2 or empty vector (negative control). Both basal and K + -stimulated glucagon secretion were inhibited by overexpression of Stmn2. Values are means +- SEM ( n = 4). * p < 0.05; ** p < 0.001 compared to unstimulated control.