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Anti ha hrp

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Anti-HA HRP is a horseradish peroxidase (HRP)-conjugated antibody that specifically recognizes the influenza hemagglutinin (HA) epitope tag. It is designed for the detection of HA-tagged proteins in various immunoassays.

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Lab products found in correlation

Anti v5 hrp, by Thermo Fisher Scientific (2 mentions) Anti flag hrp, by Merck Group (2 mentions) Ab27671, by Abcam (1 mentions) Streptavidin hrp, by Cell Signaling Technology (1 mentions) Anti ha antibody, by Cell Signaling Technology (1 mentions) Anti pmp70, by Merck Group (1 mentions) Sab4200181, by Merck Group (1 mentions) Anti myc hrp, by Cell Signaling Technology (1 mentions) Anti gfp hrp, by Cell Signaling Technology (1 mentions) Anti myc antibody, by Cell Signaling Technology (1 mentions) M20004, by Abmart (1 mentions) Anti β actin, by GenScript (1 mentions) Rabbit anti pias1 antibody, by Abcam (1 mentions) Anti v5 hrp antibody, by Thermo Fisher Scientific (1 mentions) Hrp labeled anti mouse antibody, by Jackson ImmunoResearch (1 mentions) Hrp labeled anti rabbit antibody, by Jackson ImmunoResearch (1 mentions) Anti β actin, by Merck Group (1 mentions) Protease inhibitor, by Roche (1 mentions) Bradford assay, by Bio-Rad (1 mentions) Ab157464, by Abcam (1 mentions)

5 protocols using anti ha hrp

1

Immunoblotting Antibody Detection Protocol

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Detecting reagents, including streptavidin-HRP (1:2,000 for WB, 3999S), anti-Myc antibody (1:2,000 for WB, 2276S), anti-Myc HRP (1:1,000 for WB, 2040S), anti-HA antibody (1:2,000 for WB, 2367S), anti-HA HRP (1:1,000 for WB, 2999S), V5 (1:1,000 for immunofluorescence, 13202S), and anti-GFP HRP (1:1,000 for WB, 2037S) were purchased from Cell Signaling Technology. Antibodies against V5 (1:3,000 for WB, ab27671) and MARCH5 (1:2,000 for WB, ab174959) were purchased from Abcam. Anti-GDAPH (1:4,000 for WB, A00191) and anti–β-actin (1:4,000 for WB, A00702) were purchased from GenScript. Other antibodies used in this study included anti-FLAG (1:2,000 for WB, GNI14110-FG) and anti-FLAG HRP (GNI4310-FG; GNI), anti-PMP70 (1:3,000 for WB, SAB4200181; Sigma-Aldrich), and anti-GFP (1:3,000 for WB, M20004; Abmart). Anti–V5-HRP (1:5,000 for WB, R961-25) was purchased from Thermo Fisher Scientific. Other primary antibodies used were anti-MARCH5 (19168S; Cell Signaling Technology), anti-PEX19 (14713–1-AP; Proteintech), anti-PEX3 (sc-271477; Santa Cruz Biotechnology), anti-catalase (219010; Millipore), and anti-PEX13 (ab235043; Abcam). Secondary antibodies conjugated to HRP were purchased from GenScript.
Zheng J., Chen X., Liu Q., Zhong G, & Zhuang M. (2021). Ubiquitin ligase MARCH5 localizes to peroxisomes to regulate pexophagy. The Journal of Cell Biology, 221(1), e202103156.
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2

Western Blot Analysis of Protein Targets

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Cell lysates were harvested in lysis buffer including protease inhibitors (Roche) as described previously (Lv et al., 2017 (link)). Protein concentration was determined using the Bradford assay (Biorad), and proteins were separated in SDS 4–20% polyacrylamide gels and then transferred onto a PVDF membrane. Membranes were blocked in TBS containing 5% milk, and 0.1% Tween 20 solution. Membranes were then incubated in the following primary antibodies: mouse anti-RTA (Argene, Cat# 11–008, Discontinued), anti-β-actin (Sigma, Cat # A5441), rabbit anti-PU.1 antibody (Cell Signaling Tech, Cat # 2258), rabbit anti-IRF8 antibody (Cell Signaling Tech, Cat # 5628), rabbit anti-PIAS1 antibody (Abcam, Cat # 109388), anti-HA-HRP (Cell Signaling Tech, Cat# 14031), anti-V5-HRP antibody (Thermo Fisher, Cat # R961–25) and anti-Flag-HRP antibody (Cell Signaling Tech, Cat# 86861 and Cat # 2044). The secondary antibodies (1:5,000 dilutions) used were horseradish peroxidase (HRP)-labeled anti-rabbit antibody (Jackson ImmunoResearch, Cat # 111–035-144), HRP-labeled anti-mouse antibody (Jackson ImmunoResearch, Cat # 111–035-166), HRP-labeled anti-rabbit antibody, light chain specific (Jackson ImmunoResearch, Cat # 211–032-171).
Zhang K., Lv D.W, & Li R. (2019). Protein inhibitor of activated STAT1 (PIAS1) inhibits IRF8 activation of Epstein-Barr virus lytic gene expression. Virology, 540, 75-87.
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3

Characterizing GATAD2A-CHD Protein Interactions

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Immunoprecipitation assays were performed as previously described.16 (link) In brief, HA-GATAD2AWT and HA-GATAD2AC420Y proteins were independently co-expressed with each of the three FLAG-CHD-CTD fusion proteins in rabbit reticulocyte lysates for in vitro translation (IVT). Expressed FLAG-CHD-CTD proteins together with HA-GATAD2A proteins were immobilized on anti-FLAG resin, washed, and eluted with 3X-FLAG peptide. In parallel, IVT lysates expressing only an HA-GATAD2A or FLAG-CHD-CTD protein, as well as lysates with no expressed protein, were run as negative controls. Immunoprecipitation inputs and eluates were loaded and run with sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by transfer to a polyvinylidene fluoride membrane. Immunoblots were probed with anti-HA-HRP (1:20,000, Cell Signaling Technology, no.2999S) followed by probing with anti-FLAG-HRP (1:80,000, Sigma Aldrich, no. A8592).
Werren E.A., Guxholli A., Jones N., Wagner M., Hannibal I., Granadillo J.L., Tyndall A.V., Moccia A., Kuehl R., Levandoski K.M., Day-Salvatore D.L., Wheeler M., Chong J.X., Bamshad M.J., Innes A.M., Pierson T.M., Mackay J.P., Bielas S.L, & Martin D.M. (2023). De novo variants in GATAD2A in individuals with a neurodevelopmental disorder: GATAD2A-related neurodevelopmental disorder. Human Genetics and Genomics Advances, 4(3), 100198.
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4

Chromatin Immunoprecipitation Profiling Antibodies

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The following commercially available antibodies were used:
Anti-H3 (1:5.000, #61475), Anti-H3K36me3 (1:4.000, #61102), Anti-H3K36me2 (1:1.000, #39255), Anti-H3K36me1 (1:1.000, #61351), Anti-H3K4me3 (1:500, #61379) and anti-H2A (1:1.000, #39209) were from Active Motif. Anti-H3K27me3 (1:1.000, C15410069), Anti-H3K79me3 (1:1.000, C15310068), anti-H2A.Zac (2 µg/IP, C15410202-050) and Anti-H3K9me3 (1:1.000, C15410056) were from Diagenode. Anti-H4K20me3 (1:1.000, ab9053), Anti-H3 (1:30.000, ab1791), anti-MTA1 (1:1.000, ab71153 or 3 µg/IP, ab50209), anti-CHD4 (1:1.000 or 2 µg/IP, ab70469), anti-HDAC2 (1:5.000, ab124974) anti-RBBP4 (1:1.000, ab488), anti-RBBP7 (1:1.000, ab3535), Anti-H3K27ac (2 µg/IP, ab4729), anti-H2A.Z (1:3.000, ab4174), and anti-MBD3 (1:5.000, ab157464) were from Abcam. Anti-PWWP2A (1:1.000, NBP2-13833) from Novus (Acris). Anti-HA-HRP (1:40.000, 2999S) and anti-HDAC1 (1:20.000, 5356S) from Cell Signaling Technology. Anti-FLAG M2 (1:80.000 or 3 µg/IP, F1804) and anti-mouse IgG (3 µg/IP, I8765) from Sigma-Aldrich.
The following secondary antibodies were used:
Anti-rabbit IRDye 800CW (1:10.000, 926-32211) and anti-mouse IRDye 680RD (1:10.000 926‐68070) from LI-COR Biosciences. Anti-mouse-HRP (1:10.000, M114) from Leinco Technologies. Anti-mouse and anti-rabbit HRP-linked antibodies (1:10.000, NA931 and NA934) from VWR.
Link S., Spitzer R.M., Sana M., Torrado M., Völker-Albert M.C., Keilhauer E.C., Burgold T., Pünzeler S., Low J.K., Lindström I., Nist A., Regnard C., Stiewe T., Hendrich B., Imhof A., Mann M., Mackay J.P., Bartkuhn M, & Hake S.B. (2018). PWWP2A binds distinct chromatin moieties and interacts with an MTA1-specific core NuRD complex. Nature Communications, 9, 4300.
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5

Immunoaffinity Purification and Detection

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Whole cells were lysed in ice-cold RIPA buffer containing protease inhibitor cocktails (Sigma-Aldrich, 11836145001) at 4 °C and centrifuged at max speed (15000 rpm) for 10 min. The lysates were incubated with the indicated beads (HA-antibody-conjugated beads (Thermo Scientific, #88836), Anti-V5-tag mAb-Magnetic Beads (VWR, #M167-11), or ANTI-FLAG® M2 Affinity Gel (Sigma-Aldrich, # A2220)) overnight at 4 °C. Beads were washed four times with RIPA buffer with protease inhibitor cocktails and boiled in an appropriate amount of 2× Laemmli sample buffer (Sigma-Aldrich, #S3401-10VL). Western blotting was performed as described above (n = 3). Membranes were probed with anti-V5 HRP (Thermo Fisher Scientific, #R961-25, 1:1000), anti-HA HRP (Cell Signaling Technology, #14031, 1:1000), or anti-Flag HRP (Sigma-Aldrich, #A8592, 1:3000) antibodies.
Wu X.J., Zhang Z., Wong J.P., Rivera-Soto R., White M.C., Rai A.A, & Damania B. (2023). Kaposi’s sarcoma-associated herpesvirus viral protein kinase augments cell survival. Cell Death & Disease, 14(10), 688.
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