Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker

• Supplier: Abcam
• Catalog: ab133615
• Prices: $385.00
• Sizes: 100 µl
• Host: Rabbit
• Clonality: Monoclonal
• Isotype: IgG
• Clone: EPR3633(2)
• Applications: ICC/IF ICC/IF WB IHC-P FC
• Species Reactivities: Rat, Human
• Antigen: Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Calnexin aa 550-650 (C terminal)
• Description: Rabbit Monoclonal
• Gene: CANX
• Conjugate: Unconjugated

Product images

Overlay histogram showing HeLa cells fixed in 80% methanol and stained with purified ab133615 at a dilution of 1 in 360 (red line). The secondary antibody used was FITC goat anti-rabbit at a dilution of 1 in 150. Rabbit monoclonal IgG was used as an isotype control (black line) and cells without incubation with antibody were used as a negative control (blue line).

Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (ab133615) at 1/1000 dilution (purified) + HepG2 cell lysate at 20 µgSecondaryHRP goat anti-rabbit (H+L) at 1/1000 dilution

Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (ab133615) at 1/5000 dilution (purified) + HeLa cell lysate at 20 µgSecondaryHRP goat anti-rabbit (H+L) at 1/1000 dilution

Immunohistochemical staining of paraffin embedded rat cardiac muscle with purified ab133615 at a working dilution of 1 in 4000. The secondary antibody used is ab97051, a HRP goat anti-rabbit (H+L), at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

Immunohistochemical staining of paraffin embedded human pancreas with purified ab133615 at a working dilution of 1 in 4000. The secondary antibody used is ab97051, a HRP goat anti-rabbit (H+L), at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

Immunofluorescence staining of HeLa cells with purified ab133615 at a working dilution of 1 in 200, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti rabbit, used at a dilution of 1 in 500. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative control is shown in bottom right hand panel - for the negative control, purified ab133615 was used at a dilution of 1/200 followed by an Alexa Fluor® 488 goat anti-mouse antibody at a dilution of 1/500.

Overlay histogram showing HeLa cells stained with unpurified ab133615 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (unpurified ab133615, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

All lanes : Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (ab133615) at 1/1000 dilution (unpurified)Lane 1 : HepG2 cell lysateLane 2 : A431 cell lysateLane 3 : SH-SY5Y cell lysateLane 4 : HeLa cell lysateLysates/proteins at 10 µg per lane.SecondaryHRP labelled goat anti-rabbit at 1/2000 dilution

Immunohistochemical analysis of paraffin-embedded human kidney tissue labelled with unpurified ab133615 at 1/100 dilution.