MA5-45403

antibody from Invitrogen Antibodies

Targeting: HCN2 BCNG-2, BCNG2, HAC-1

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunoprecipitation (Recommended by provider)

Immunohistochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-45403

Provider

Invitrogen Antibodies

Product name

HCN2 Monoclonal Antibody (S71), APC

Antibody type

Monoclonal

Antigen

Other

Description

1 µg/mL of MA5-45403 was sufficient for detection of HCN2 in 10 µg of rat brain lysate by colorimetric immunoblot analysis using Goat anti-mouse IgG:HRP as the secondary antibody.|Detects approximately 95kDa. No cross-reactivity against HCN1.

Reactivity

Human, Mouse, Rat

Host

Mouse

Isotype

IgG

Antibody clone number

S71

Vial size

100 µg

Concentration

1 mg/mL

Storage

4° C

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot analysis of Rat brain membrane lysate showing detection of HCN2 protein. Load: 15 µg. Blocking: 1.5% BSA for 30 minutes at RT. Samples were incubated with HCN2 monoclonal antibody (Product # MA5-45403) at 1:1,000 for 2 hours at RT, followed by Sheep Anti-Mouse IgG: HRP for 1 hour at RT.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot analysis of Mouse Brain showing detection of ~95 kDa HCN2 protein. Lane 1: MW Ladder. Lane 2: Mouse Brain (15 µg). Load: 15 µg. Blocking: 5% Skim Milk powder in TBST. Samples were incubated with HCN2 monoclonal antibody (Product # MA5-45403) at 1:1,000 for 2 hours at RT with shaking, followed by Goat anti-mouse IgG:HRP at 1:4,000 for 1 hour at RT with shaking. Color Development: Chemiluminescent for HRP (Moss) for 5 min in RT. Predicted/Observed Size: ~95 kDa.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunocytochemistry/Immunofluorescence analysis using differentiated SH-SY5Y cells. Samples were incubated with HCN2 monoclonal antibody (Product # MA5-45403) at 1:100, followed by AlexaFluor 488. Counterstain used was phalloidin (Alexa 647, red), beta tubulin (Anti-beta III Tubulin Ab, Alexa 555, magenta) Hoechst (blue). (A) Phalloidin (B) Anti-beta III Tubulin Ab. (C) HCN2 antibody. (D) Hoechst (E) Composite.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunocytochemistry/Immunofluorescence analysis using human neuroblastoma cells. Fixation involved 4% PFA for 15 min. Samples were incubated with HCN2 monoclonal antibody (Product # MA5-45403) at 1:50 for overnight at 4°C with slow rocking, followed by AlexaFluor 488 at 1:1,000 for 1 hour at RT. Counterstain used was Phalloidin-iFluor 647 (red) F-Actin stain; Hoechst (blue) nuclear stain at 1:800, 1.6mM for 20 min at RT. (A) Hoechst (blue) nuclear stain. (B) Phalloidin-iFluor 647 (red) F-Actin stain. (C) HCN2 antibody (D) Composite.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis using hippocampus. Fixation involved Bouins Fixative and paraffin-embedded. Samples were incubated with HCN2 monoclonal antibody (Product # MA5-45403) at 1:100 for 1 hour at RT, followed by FITC Goat Anti-Mouse (green) at 1:50 for 1 hour at RT.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis using frozen brain section. Fixation involved 10% formalin solution for 12-24 hours at RT. Samples were incubated with HCN2 monoclonal antibody (Product # MA5-45403) at 1:1,000 for 1 hour at RT, followed by HRP/DAB Detection System: Biotinylated Goat Anti-Mouse, Streptavidin Peroxidase, DAB Chromogen (brown) for 30 minutes at RT. Counterstain used was Mayer Hematoxylin (purple/blue) nuclear stain at 250-500 µL for 5 minutes at RT.