702260

antibody from Invitrogen Antibodies

Targeting: FKBP5 FKBP51, FKBP54, P54, PPIase, Ptg-10

Antibody data

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Validation data

Product number: 702260 - Provider product page
Provider: Invitrogen Antibodies
Product name: FKBP5 Recombinant Rabbit Monoclonal Antibody (6H8L4)
Antibody type: Monoclonal
Antigen: Other
Reactivity: Human, Mouse, Rat
Host: Rabbit
Isotype: IgG
Antibody clone number: 6H8L4
Vial size: 100 µg
Concentration: 0.5 mg/mL
Storage: Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

FKBP5 protein structure - 702260 shown in red.

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), HT-29 (Lane 2), HCT 116 (Lane 3), HEL 92.1.72. (Lane 4), HEK-293 (Lane 5), Jurkat (Lane 6) and K-562 (Lane 7). The blots were probed with Anti-FKBP5 Recombinant Rabbit Monoclonal Antibody (Product # 702260, 1-2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 51 kDa band corresponding to EAAT2 was observed across cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of FKBP5 was achieved by transfecting HeLa with FKBP5 specific siRNAs (Silencer® select Product # S5214, S5216). Western blot analysis (Fig. A) was performed using Whole cell extracts from the FKBP5 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with FKBP5 Antibody (6H8L4), ABfinity Rabbit Monoclonal (Product # 702260, 1 µg/mL) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000). 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 FKBP5.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-FKBP5 Antibody (6H8L4), ABfinity Rabbit Monoclonal (Product # 702260) and a 51kDa band corresponding to FKBP5 was observed across all the tested cell lines. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), HT-29 (Lane 2), HCT 116 (Lane 3), PC-3 (Lane 4), HEK-293 (Lane 5), Jurkat (Lane 6), Neuro-2a (Lane 7), PC-12 (Lane 8) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0302BOX). 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 µg/mL) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis was performed on fixed and permeabilized PC-3 cells for detection of endogenous FKBP5 using Anti-FKBP5 Recombinant Rabbit Monoclonal Antibody (Product # 702260, 2 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Panel a) shows representative cells that were stained for detection and localization of FKBP5 protein (green), Panel b) is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). Panel c) represents cytoskeletal F-actin staining using Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d) is a composite image of Panels a, b and c clearly demonstrating localization of FKBP5 in the cytoplasm. 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: Immunofluorescence analysis was performed on fixed and permeabilized HeLa cells for detection of endogenous FKBP5 using Anti-FKBP5 Recombinant Rabbit Monoclonal Antibody (Product # 702260, 2 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Panel a) shows representative cells that were stained for detection and localization of FKBP5 protein (green), Panel b) is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). Panel c) represents cytoskeletal F-actin staining using Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d) is a composite image of Panels a, b and c clearly demonstrating predominantly nuclear localization of FKBP5. Panel e) represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.