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Western blotAntibody data
- Antibody Data
- Antigen structure
- References [0]
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- Validations
- Western blot [3]
- Immunocytochemistry [1]
- Immunohistochemistry [8]
- Chromatin Immunoprecipitation [1]
- Other assay [1]
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- Product number
- PA5-88090 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- POLR2A Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- Positive Samples: HeLa, BT-474, H460, Jurkat, Mouse testis, Mouse brain; Cellular Location: Nucleus Immunogen sequence: MHGGGPPSGD SACPLRTIKR VQFGVLSPDE LKRMSVTEGG IKYPETTEGG RPKLGGLMDP RQGVIERTGR CQTCAGNMTE CPGHFGHIEL AKPVFHVGFL VKTMKVLRCV CFFCSKLLVD SNNPKIKDIL AKSKGQPKKR LTHVYDLCKG KNICEGGEEM DNKFGVEQPE GDEDLTKEKG HGGCGRYQPR IRRSGLELYA EWKHVNEDSQ EKKILLSPER VHEIFKRISD EECFVLGMEP RYARPEWMIV TVLPVPPLSV
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 0.47 mg/mL
- Storage
- -20° C, Avoid Freeze/Thaw Cycles
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of extracts of various cell lines, using POLR2A Polyclonal antibody (Product # PA5-88090) at 1:1000 dilution. Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) at 1:10000 dilution. Lysates/proteins: 25ug per lane. Blocking buffer: 3% nonfat dry milk in TBST. Exposure time: 30s.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot analysis of POLR2A in extracts of various cell lines using POLR2A Polyclonal Antibody (Product # PA5-88090) at a dilution of 1:1000. A HRP Goat Anti-Rabbit IgG (H+L) secondary antibody was used at a dilution of 1:10,000. Lysates/proteins: 25 µg per lane. Blocking buffer: 3% nonfat dry milk in TBST.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-POLR2A Polyclonal Antibody (Product # PA5-88090) and a 250 kDa band along with uncharacterized band (*) at ~35 kDa corresponding to POLR2A was observed across cell lines tested. Whole cell extracts (40 µg lysate) of HeLa (Lane 1), MCF7 (Lane 2), Jurkat (Lane 3), HCT 116 (Lane 4), DU 145 (Lane 5), LNCaP (Lane 6), H-1975 (Lane 7) and Raji (Lane 8) were electrophoresed using NuPAGE™ 3-8% Tris-Acetate Protein Gel (Product # EA0378BOX). Resolved proteins were equilibrated with 20% ethanol and then transferred onto a nitrocellulose membrane (Product # IB23002) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:20000 dilution using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Pico PLUS Chemiluminescent Substrate (Product # 34580).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of POLR2A Polyclonal Antibody was performed using 70% confluent log phase HCT 116 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 45 minutes at room temperature. The cells were labeled with POLR2A Polyclonal Antibody (Product # PA5-88090) at 1:100 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 dilution). Panel d represents the merged image showing Nuclear 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 analysis of POLR2A in paraffin-embedded rat brain using POLR2A Polyclonal Antibody (Product # PA5-88090) at a dilution of 1:100.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of POLR2A in paraffin-embedded mouse brain using POLR2A Polyclonal Antibody (Product # PA5-88090) at a dilution of 1:100.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of POLR2A in paraffin-embedded rat testis using POLR2A Polyclonal Antibody (Product # PA5-88090) at a dilution of 1:100.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of POLR2A in paraffin-embedded rat brain using POLR2A Polyclonal Antibody (Product # PA5-88090) at a dilution of 1:100.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of POLR2A in paraffin-embedded rat pancreas using POLR2A Polyclonal Antibody (Product # PA5-88090) at a dilution of 1:100.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of POLR2A in paraffin-embedded mouse testis using POLR2A Polyclonal Antibody (Product # PA5-88090) at a dilution of 1:100.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of POLR2A in paraffin-embedded mouse brain using POLR2A Polyclonal Antibody (Product # PA5-88090) at a dilution of 1:100.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of POLR2A in paraffin-embedded mouse pancreas using POLR2A Polyclonal Antibody (Product # PA5-88090) at a dilution of 1:200.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Chromatin Immunoprecipitation (ChIP) was performed using POLR2A Polyclonal Antibody (Product # PA5-88090, 5 µg) on sheared chromatin from a million HCT 116 cells using the MAGnify ChIP System (Product # 49-2024). Normal Rabbit IgG was used as a negative IP control. The purified DNA was analyzed by qPCR with PCR primer pairs over Myc and GAPDH genes (active), and MYOD1 and Myc (+11382) (inactive). Antibody specificity was demonstrated by detection of enrichment of the target protein at specific gene loci. Data is presented as fold enrichment of the antibody signal versus the Rabbit Isotype using the comparative CT method.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunoprecipitation analysis of POLR2A was performed in 200 µg extracts of HeLa cells using POLR2A Polyclonal Antibody (Product # PA5-88090). Western blot was performed from the immunoprecipitate using POLR2A Polyclonal Antibody at a dilution of 1:1000.
