Anti-Calreticulin antibody

• Supplier: Abcam
• Catalog: ab4
• Prices: $400.00
• Sizes: 100 µl
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: IgG
• Applications: ICC/IF ICC/IF IHC-P WB IP ICC/IF
• Species Reactivities: Mouse, Rat, Chicken, Hamster, Bovine, Human, Zebrafish, Monkey
• Antigen: Fusion protein, containing amino acids 154-347 of Mouse Calreticulin
• Description: Rabbit Polyclonal
• Gene: CALR
• Conjugate: Unconjugated

Product images

All lanes : Anti-Calreticulin antibody (ab4) at 1/1000 dilutionLane 1 : Chicken lymphoid cell line lysateLane 2 : Chicken lymphoid cell line lysateLysates/proteins at 1000000 cells per lane.SecondaryPolyclonal HRP-conjugate Goat anti-rabbit IgG at 1/2000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.

IF staining was performed on HCT15 colon cancer cells using a 1:50 dilution of the primary antibody followed by hybridisation with a 1:100 dilution of the secondary anti-rabbit antibody Alexa Fluor 488 (molecular Probes) with green fluorescence. Picture from review by Karin Birkenkamp-Demtroeder submitted 20 July 2004

ICC/IF image of ab4 stained HepG2 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab4, 1/1000 dilution) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

ab4 staining calreticulin in Human placenta.Left panel: with primary antibody diluted 1:4000. Right panel: isotype control.Sections were stained using an automated system (DAKO Autostainer Plus), at room temperature: sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffers EDTA pH 9.0. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.

Product References

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