Explore
Validate
Learn
Western blotAntibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [5]
- Immunocytochemistry [2]
- Immunohistochemistry [5]
Submit
Validation data
Reference
Comment
Report error
- Product number
- PA5-22281 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- HPRT1 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant protein fragment
- Description
- Recommended positive controls: 293T, A431, HeLa, HepG2, Molt-4, Raji, mouse liver.
- Concentration
- 1.06 mg/mL
No comments: Submit comment
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of HPRT using 30 µg of A) MOLT4 and B) Raji lysate. Samples were loaded onto a 12% SDS-PAGE gel and probed with a HPRT polyclonal antibody (Product # PA5-22281) at a dilution of 1:1000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot analysis of HPRT1 was performed by separating 30 µg of various whole cell extracts by 12% SDS-PAGE. Proteins were transferred to a membrane and probed with a HPRT1 Polyclonal Antibody (Product # PA5-22281) at a dilution of 1:1000 and a HRP-conjugated anti-rabbit IgG secondary antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- HPRT1 Polyclonal Antibody detects HPRT1 protein by western blot analysis. A. 50 µg mouse liver lysate/extract.12% SDS-PAGE. HPRT1 Polyclonal Antibody (Product # PA5-22281) dilution: 1:500. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of HPRT1 was performed by loading 30 µg of ME-180 CAS9 control (Lane 1) and ME-180 - HPRT1 knockout (Lane 2) membrane extracts. The blot was probed with HPRT1 Polyclonal Antibody (Product # PA5-22281, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL 1:4000 dilution). Loss of signal upon CRISPR mediated knockout (KO) confirms that antibody is specific to HPRT1.
- 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), HT-29 (Lane 2), SW480 (Lane 3), A549 (Lane 4), PC-3 (Lane 5), DU 145 (Lane 6), MCF7 (Lane 7), Hep G2 (Lane 8) and A-431 (Lane 9). The blot was probed with Anti- HPRT1 Polyclonal Antibody (Product # PA5-22281, 1:1000 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 24 kDa band corresponding to HPRT1 was observed across the cell lines tested.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of HPRT in paraformaldehyde-fixed HeLa cells using a HPRT polyclonal antibody (Product # PA5-22281) at a 1:200 dilution.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of HPRT1 was performed using 70% confluent log phase HeLa cells. 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 HPRT1 Rabbit Polyclonal Antibody (Product # PA5-22281) at 5 microgram/mL 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® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415). Panel d represents the merged image showing Cytoplasm and Membrane localization. Panel e 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 (Paraffin) analysis of HPRT1 was performed in paraffin-embedded human colon cancer tissue using HPRT1 Polyclonal Antibody (Product # PA5-22281) at a dilution of 1:500.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry (Paraffin) analysis of HPRT1 was performed in paraffin-embedded human endometrial cancer tissue using HPRT1 Polyclonal Antibody (Product # PA5-22281) at a dilution of 1:500.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry (Paraffin) analysis of HPRT1 was performed in paraffin-embedded rat brain tissue using HPRT1 Polyclonal Antibody (Product # PA5-22281) at a dilution of 1:500.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry (Paraffin) analysis of HPRT1 was performed in paraffin-embedded mouse colon tissue using HPRT1 Polyclonal Antibody (Product # PA5-22281) at a dilution of 1:500.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry (Paraffin) analysis of HPRT1 was performed in paraffin-embedded rat colon tissue using HPRT1 Polyclonal Antibody (Product # PA5-22281) at a dilution of 1:500.
