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- Antibody Data
- Antigen structure
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- Validations
- Western blot [2]
- Immunocytochemistry [2]
- Immunohistochemistry [1]
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- Product number
- PA5-53890 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- AHNAK Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant full-length protein
- Description
- Immunogen sequence: LGEGHLSVKG SGGEWKGPQV SSALNLDTSK FAGGLHFSGP KVEGGVKGGQ IGLQAPGLSV SGPQGHLESG SGKVTFPKMK IPKFTFSGRE LVGREMGVDV HFPKAEASIQ AGAGDGEWEE SEVKLKKSKI KMPK
- Concentration
- 0.1 mg/mL
Submitted references Discovery of Spatial Peptide Signatures for Neuroblastoma Risk Assessment by MALDI Mass Spectrometry Imaging.
Wu Z, Hundsdoerfer P, Schulte JH, Astrahantseff K, Boral S, Schmelz K, Eggert A, Klein O
Cancers 2021 Jun 25;13(13)
Cancers 2021 Jun 25;13(13)
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of AHNAK was achieved by transfecting HaCaT with AHNAK specific siRNAs (Silencer® select Product # S41427, S41428). Western blot analysis (Fig. a) was performed using whole cell extracts from the AHNAK knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with AHNAK Polyclonal Antibody (Product # PA5-53890, 1:2000) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:20,000) and detected by chemiluminescence using the iBright™ FL1500 Imaging System (Product # A44115). 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 AHNAK.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using AHNAK Polyclonal Antibody (Product # PA5-53890) and a 680 kDa band corresponding to AHNAK was observed across cell lines tested. Whole cell extracts (60 µg lysate) prepared with RIPA buffer, of U-87 MG (Lane 1), PANC-1 (Lane 2), 769-P (Lane 3), NIH:OVCAR-3 (Lane 4), HaCaT (Lane 5), SW480 (Lane 6), Jurkat (Lane 7), A-673 (Lane 8), HEL 92.1.7 (Lane 9) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX), 10 well. Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23002) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:2000) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:20,000) using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Atto Ultimate Sensitivity Substrate (Product # A38556).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent staining of AHNAK in human cell line U-2 OS shows positivity in plasma membrane & cytoplasm. Samples were probed using an AHNAK Polyclonal Antibody (Product # PA5-53890).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of AHNAK was performed using 70% confluent log phase HaCaT cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with AHNAK Polyclonal Antibody (Product # PA5-53890, 0.25 µg/mL) in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor™ Plus 488 (Product # A32790, 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, 1:300). Panel d represents the merged image showing cell membrane and cytosolic 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
- Immunohistochemical staining of AHNAK in human oral mucosa shows cytoplasmic positivity in squamous epithelial cells. Samples were probed using an AHNAK Polyclonal Antibody (Product # PA5-53890).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure 3 Validation of two discriminative protein markers for neuroblastoma risk in tissue sections. Shown are representative tissue sections from neuroblastoma designated high-risk (HR) and as other risk groups (nHR). MALDI-MSI ion maps for one peptide (m/z 1832.79 Da) assigned to AHNAK and one peptide (m/z 922.50 Da) assigned to CRMP1 are shown next to the corresponding sections stained with hematoxylin and eosin (H&E) for orientation. Black lines border areas with >80% tumor cell content. Immunohistochemical (IHC) detection of AHNAK and CRMP1 is shown for the regions surrounded by the yellow squares in the expanded image (400x magnification).
