MA5-45557

antibody from Invitrogen Antibodies

Targeting: CLCN3 ClC-3, CLC3

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Recommended by provider)

Antibody Data

Product number

MA5-45557

Provider

Invitrogen Antibodies

Product name

CLCN3 Monoclonal Antibody (N258/5), PE

Antibody type

Monoclonal

Antigen

Synthetic peptide

Description

1 µg/mL of MA5-45557 was sufficient for detection of Clcn3 in 20 µg of rat brain membrane lysate and assayed by colorimetric immunoblot analysis using goat anti-mouse IgG:HRP as the secondary antibody.|Detects approximately 90kDa. Does not cross-react with Clcn4 or Clcn5 (based on KO validation results). This antibody was formerly sold as clone S258-5.

Reactivity

Human, Mouse, Rat

Host

Mouse

Conjugate

Yellow dye

Isotype

IgG

Antibody clone number

N258/5

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 showing detection of ~90 kDa CIcn3 protein. Lane 1: Molecular Weight Ladder. Lane 2: Rat Brain Membrane. Load: 15 µg. Blocking: 2% BSA and 2% Skim Milk in 1X TBST. Samples were incubated with CIcn3 monoclonal antibody (Product # MA5-45557) at 1:200 for 16 hours at 4°C, followed by Goat Anti-Mouse IgG: HRP at 1:1,000 for 1 hour RT. Color Development: ECL solution for 6 min in RT. Predicted/Observed Size: ~90 kDa.

Immunocytochemistry

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 Clcn3 monoclonal antibody (Product # MA5-45557) 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) Clcn3 antibody (D) Composite.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunocytochemistry/Immunofluorescence analysis using human neuroblastoma cells. Fixation involved 4% Formaldehyde for 15 min at RT. Samples were incubated with Clcn3 monoclonal antibody (Product # MA5-45557) at 1:100 for 60 min at RT, followed by Goat Anti-Mouse ATTO 488 at 1:100 for 60 min at RT. Counterstain used was Phalloidin Texas Red F-Actin stain; DAPI (blue) nuclear stain at 1:1,000, 1:5,000 for 60 min RT, 5 min RT. Localization: Membrane, Endosome, Endosome membrane, Cytoplasmic Vesicle, Secretory Vesicle Membrane. Magnification: 60X. (A) DAPI (blue) nuclear stain. (B) Phalloidin Texas Red F-Actin stain. (C) Clcn3 antibody. (D) Composite.

Conjugate

Yellow dye

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunocytochemistry/Immunofluorescence analysis using human neuroblastoma cells. Fixation involved 4% Formaldehyde for 15 min at RT. Samples were incubated with Clcn3 monoclonal antibody (Product # MA5-45557) at 1:100 for 60 min at RT, followed by Goat Anti-Mouse ATTO 488 at 1:100 for 60 min at RT. Counterstain used was Phalloidin Texas Red F-Actin stain; DAPI (blue) nuclear stain at 1:1,000, 1:5,000 for 60 min RT, 5 min RT. Localization: Membrane, Endosome, Endosome membrane, Cytoplasmic Vesicle, Secretory Vesicle Membrane. Magnification: 60X. (A) DAPI (blue) nuclear stain. (B) Phalloidin Texas Red F-Actin stain. (C) Clcn3 antibody. (D) Composite.