PA5-29780
antibody from Invitrogen Antibodies
Targeting: TNFRSF14
ATAR, CD270, HVEA, HVEM, LIGHTR, TR2
Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [6]
- Immunocytochemistry [1]
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Validation data
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- Product number
- PA5-29780 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- TNFRSF14 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant protein fragment
- Description
- Recommended positive controls: A431, Molt-4, mouse spleen. Predicted reactivity: Rhesus Monkey (82%). Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 0.78 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 membrane extracts (30 µg lysate) of KARPAS 299 (Lane 1), Ramos (Lane 2), Jurkat (Lane 3), MOLT-4 (Lane 4), A-431 (Lane 5), tissue extracts of Mouse Spleen (Lane 6) and Rat Spleen (Lane 7). The blot was probed with Anti-TNFRSF14 Rabbit Polyclonal Antibody (Product # PA5-29780, 1µg/mL) 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 30 kDa band corresponding to TNFRSF14 was detected across the cell lines and tissues tested.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using TNFRSF14 Polyclonal Antibody (Product # PA5-29780). Mouse tissue extract (30 µg) was separated by 10% SDS-PAGE, and the membrane was blotted with TNFRSF14 Polyclonal Antibody (Product # PA5-29780) diluted at 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 TNFRSF14 was performed by separating 30 µg of non-transfected (–) and transfected (+) 293T whole cell extracts by 10% SDS-PAGE. Proteins were transferred to a membrane and probed with a TNFRSF14 Polyclonal Antibody (Product # PA5-29780) at a dilution of 1:5000. 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 TNFRSF14 was performed by separating 30 µg of whole cell extract by 10% SDS-PAGE. Proteins were transferred to a membrane and probed with a TNFRSF14 Polyclonal Antibody (Product # PA5-29780) at a dilution of 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 TNFRSF14 was performed by separating 30 µg of whole cell extract by 10% SDS-PAGE. Proteins were transferred to a membrane and probed with a TNFRSF14 Polyclonal Antibody (Product # PA5-29780) at a dilution of 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
- Knockdown of TNFRSF14 was achieved by transfecting A431 with TNFRSF14 specific siRNAs (Silencer® select Product # s16700, s16699). Western blot analysis (Fig. a) was performed using whole cell extracts from the TNFRSF14 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with TNFRSF14 Polyclonal Antibody (Product # PA5-29780, 1:250 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). Reduction of signal upon siRNA mediated knock down confirms that antibody is specific to TNFRSF14.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of TNFRSF14 was performed using log phase K-562 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with TNFRSF14 Rabbit Polyclonal Antibody (Product # PA5-29780) at 5 µg/mL in 0.1% BSA and incubated overnight at 4 degree 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 SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing membrane localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.