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Western blot
ImmunoprecipitationAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [3]
- Immunocytochemistry [1]
- Immunohistochemistry [1]
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- Product number
- PA1-30197 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Calnexin Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- Positive control of heat shocked HeLa Cell lysate suggested.
- Reactivity
- Human, Mouse, Rat, Bovine, Chicken/Avian, Guinea Pig, Hamster, Porcine, Rabbit, Xenopus
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 1 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on whole cell extracts (30 µg lysate) of HeLa (Lane 1), HCT116 (Lane 2), A431 (Lane 3), HepG2 (Lane 4), Raji (Lane 5), MOLT-4 (Lane 6), Jurkat (Lane 7) and RAW 264.7 (Lane 8). The blot was probed with Anti- Calnexin Polyclonal Antibody (Product # PA1-30197, 1:5000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/ml, 1:4000 dilution). A 72 kDa band corresponding to Calnexin was observed across all the cell lines tested.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis using Calnexin Polyclonal Antibody (Product # PA1-30197): Lane 1: MWM, Lane 2: Vero, Lane 3: 3T3, Lane 4: PC-12, Lane 5: HeLa.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of Calnexin was achieved by transfecting HeLa cells with Calnexin specific siRNAs (Silencer® select Product # s2376). Western blot analysis (Fig. a) was performed using whole cell extracts from the Calnexin knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with Calnexin Polyclonal Antibody (Product # PA1-30197, 1:5000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25µg/ml, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to Calnexin.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of Calnexin was performed using 70% confluent log phase HeLa cells treated with Thapsigargin (1uM for 24hrs). The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Calnexin Polyclonal Antibody (Product # PA1-30197) at 1:200 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 594 conjugate (Product # A-11062) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: red). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: green) was stained with Alexa Fluor™ 488 Phalloidin (Product # A12379, 1:300). Panel d represents the merged image showing Calnexin in the ER and cytoplasm. Panel e represents the untreated cells showing lower expression levels. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of human placenta tissue stained with Calnexin Polyclonal Antibody (Product # PA1-30197) at 5 µg/mL.
