Antibody data
- Antibody Data
- Antigen structure
- References [1]
- Comments [0]
- Validations
- Western blot [4]
- Immunocytochemistry [4]
- Immunohistochemistry [2]
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Validation data
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- Product number
- MA5-32421 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- HSPB8 Recombinant Rabbit Monoclonal Antibody (JJ08-53)
- Antibody type
- Monoclonal
- Antigen
- Synthetic peptide
- Description
- Recombinant rabbit monoclonal antibodies are produced using in vitro expression systems. The expression systems are developed by cloning in the specific antibody DNA sequences from immunoreactive rabbits. Then, individual clones are screened to select the best candidates for production. The advantages of using recombinant rabbit monoclonal antibodies include: better specificity and sensitivity, lot-to-lot consistency, animal origin-free formulations, and broader immunoreactivity to diverse targets due to larger rabbit immune repertoire.
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Antibody clone number
- JJ08-53
- 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.
Submitted references HSPB8 overexpression prevents disruption of blood-brain barrier after intracerebral hemorrhage in rats through Akt/GSK3β/β-catenin signaling pathway.
Hou Y, Hu Z, Gong X, Yang B
Aging 2020 Sep 4;12(17):17568-17581
Aging 2020 Sep 4;12(17):17568-17581
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of HSPB8 in different lysates using a Monoclonal antibody (Product #MA5-32421) at a dilution of 1:1,000. Positive control: Lane 1: HepG2, Lane 2: Mouse skeletal muscle.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of HSPB8 in different lysates using a Monoclonal antibody (Product #MA5-32421) at a dilution of 1:1,000. Positive control: Lane 1: HepG2, Lane 2: Mouse skeletal muscle.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of HSPB8 was achieved by transfecting HeLa with HSPB8 specific siRNAs (Silencer® select Product # s25418, s25420). Western blot analysis (Fig. a) was performed using whole cell extracts from the HSPB8 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with HSPB8 Recombinant Rabbit Monoclonal Antibody (JJ08-53) (Product # MA5-32421, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:10000 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 Heat shock protein beta-8.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-HSPB8 Recombinant Rabbit Monoclonal Antibody (JJ08-53) (Product # MA5-32421) and a 22kDa band corresponding to HSPB8 was observed in U-2 OS and HeLa in comparison to PC-3, Hep G2 and LNCaP and showed upregulation upon beta-estradiol treatment in MCF7 cells. Whole cell extracts (30 µg lysate) of U-2 OS (Lane 1), HeLa (Lane 2), PC-3 (Lane 3), Hep G2 (Lane 4), LNCaP (Lane 5), untreated MCF7 (Lane 6), beta-estradiol treated MCF7 (Lane 7) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1500 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:8000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Reagent Kit (Product # WP20005).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemical analysis of HSPB8 in A431 cells using a HSPB8 Monoclonal antibody (Product # MA5-32421) as seen in green. The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemical analysis of HSPB8 in SH-SY-5Y cells using a HSPB8 Monoclonal antibody (Product # MA5-32421) as seen in green. The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemical analysis of HSPB8 in Hela cells using a HSPB8 Monoclonal antibody (Product # MA5-32421) as seen in green. The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of HSPB8 was performed using 80% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with HSPB8 Recombinant Rabbit Monoclonal Antibody (JJ08-53) (Product # MA5-32421) at 1:250 dilution 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 dilution), 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 cytoplasmic localization of HSPB8 in HeLa cells. 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 analysis of HSPB8 of paraffin-embedded Mouse brain tissue using a HSPB8 Monoclonal antibody (Product #MA5-32421). Counter stained with hematoxylin.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of HSPB8 of paraffin-embedded rat skeletal muscle tissue using a HSPB8 Monoclonal antibody (Product #MA5-32421). Counter stained with hematoxylin.