Invitrogen

IBA1 Polyclonal Antibody

Advanced Verification

This Antibody was verified by Knockout to ensure that the antibody binds to the antigen stated.

12 Published Figures

9 References

View all (34) IBA1 antibodies

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Protocols

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IBA1 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of IBA1 was performed using 70% confluent log phase THP-1 cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with IBA1 Polyclonal Antibody (Product # PA5-21274) at 1:200 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32766, 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 membrane and cytoplasmic localization. Panel e represents HeLa cells having no expression of IBA1. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

IBA1 Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry (Paraffin) analysis of IBA1 was performed in paraffin-embedded mouse brain tissue using IBA1 Polyclonal Antibody (Product # PA5-21274) at a dilution of 1:500. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.

IBA1 Antibody in Immunohistochemistry (Frozen) (IHC (F)) — Immunohistochemistry (Frozen) analysis of IBA1 was performed in frozen sectioned E13.5 Rat brain tissue using IBA1 Polyclonal Antibody (Product # PA5-21274) at a dilution of 1:250 (Green). Red: beta Tubulin 3/ TUJ1, a mature neuron marker, stained by beta Tubulin 3/ TUJ1 antibody diluted at 1:500. Blue: Fluoroshield with DAPI.

IBA1 Antibody in Western Blot (WB) — Western blot was performed using Anti-IBA1 Polyclonal Antibody (Product # PA5-21274) and a 17 kDa band corresponding to IBA1 was observed across U-937, U-937 treated with Retinoic Acid and THP-1. Whole Cell Extract-WCL (30 µg lysate) of U-937 (Lane 1), U-937 treated with Retinoic Acid (10 µM for 24 hours) (Lane 2), THP-1 (Lane 3), A-431 (Lane 4), HeLa (Lane 5) and MCF7 (Lane 6) were electrophoresed using NuPAGE™ 12% Bis-Tris Protein Gel (Product # NP0342BOX). 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 (1:2000 dilution) 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).

IBA1 Antibody in Flow Cytometry (Flow) — Flow Cytometry analysis of S100A7 was performed in THP-1 cells using S100A7 Monoclonal Antibody (47C1068) (Product # MA1-91555) (red) at a dilution of 1:50. Black: Unlabelled sample was used as a control. Acquisition of 20,000 events were collected using a Dylight 488-conjugated secondary antibody for FACS analysis.

IBA1 Antibody — Antibody specificity was demonstrated by CRISPR-Cas9 mediated knockout of target protein. A loss of signal was observed for target protein in IBA1 KO cell line compared to control cell line using Anti-IBA1 Polyclonal Antibody (Product # PA5-21274). {KO}

IBA1 Antibody in Immunocytochemistry, Immunohistochemistry (ICC/IF, IHC) — Fig. 3 Decreased neuronal METTL3 and m 6 A in the hippocampus by AAV-shMettl3. (A) Highly conserved gene sequences of shRNA targeting region in mouse and rat Mettl3 mRNA. (B) Knockdown effect of shMettl3 was confirmed in mouse N2a cells 3 days after transient transfection (Data are means+-SEM from 3 independent experiments). (C) AAV-mediated gene delivery in the hippocampus was confirmed by immunostaining for GFP. WT C57BL/6 mice receiving bilaterally stereotaxic injections of AAV encoding for eGFP plus anti-Mettl3 shRNA (shMettl3) or scrambled control shRNA (shCtrl) into the hippocampus at 2 months of age were analyzed 1 month later. (D-F) Representative images of the AAV-infected areas (GFP-positive) in hippocampus for different cell markers by immunocytochemistry after fixation: MAP 2 for neurons (D), GFAP for astrocytes (E), and Iba1 for microglia (F). (G, H) Representative image of immunoreactivity of METTL3 or m 6 A (G) in AAV-shRNA-injected mice and quantification analysis (H) confirmed that METTL3 protein and m 6 A modification level were reduced in mice injected with shMettl3 virus (n = 5-7 mice per group). GFP staining has been performed in adjacent brain slices at the same time and immunoreactivities of m 6 A and METTL3 were measured in GFP-positive regions. (Data are means+-SEM, * p < 0.05, **p < 0.01, (B, H) unpaired student's t-test)

IBA1 Antibody in Immunohistochemistry (IHC) — Additional file 3: Supplementary Fig. 3. Validation of intracranial injection into hippocampus by needle track and induced neuroinflammation by METTL3 depletion in mouse hippocampus. (A) A representative image of needle track (arrow) of AAV-injected mice was shown. GFP immunoreactivity was detected in area adjacent to the needle tack. NeuN staining revealed severe neuronal loss around injected areas only in AAV-shMettl3 injected mice but not AAV-shCtrl injected mice. (B-E) Representative images of immunohistochemistry for Iba1 (B) and GFAP (Thermofisher, D) in hippocampal CA1/2 or CA3 areas in shRNA-injected mice and their quantification (C for astrocyte and E for microglia) analysis showed that METTL3 knockdown caused neuroinflammation in mouse hippocampus. (n = 4-7, *p < 0.5, **p < 0.01; C, E, unpaired student's t-test).

Product Details

PA5-21274

Applications	Tested Dilution	Publications
Western Blot (WB)	1:500-1:10,000	View 2 publications 2 publications
Immunohistochemistry (IHC)	Assay-dependent	View 6 publications 6 publications
Immunohistochemistry (Paraffin) (IHC (P))	1:100-1:1,000	-
Immunohistochemistry (Frozen) (IHC (F))	1:100-1:1,000	-
Immunocytochemistry (ICC/IF)	1:100-1:1,000	View 1 publication 1 publication
Flow Cytometry (Flow)	1:50-1:200	-

Product Specifications
Species Reactivity	Human, Mouse, Rat
Published species	Mouse
Host/Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	Synthetic peptide corresponding to a region within amino acids 85 and 147 of Iba1 (Uniprot ID#P55008) View antigen
Conjugate	Unconjugated
Form	Liquid
Concentration	0.06 mg/mL
Purification	Antigen affinity chromatography
Storage buffer	PBS, pH 7, with 20% glycerol, 1% BSA
Contains	0.025% Proclin 300
Storage conditions	Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
RRID	AB_11156585

Product Specific Information

Recommended positive controls: K562, THP-1, HL-60, mouse brain, mouse liver, rat liver, Mouse brain.

Predicted reactivity: Mouse (100%), Rat (100%), Pig (100%), Rhesus Monkey (100%).

Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.

Target Information

AIF1 is induced by cytokines and interferon. It is thought to be involved in the negative regulation of growth of vascular smooth muscle cells, which contributes to the anti-inflammatory response to vessel wall trauma.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

METTL3-dependent RNA m⁶A dysregulation contributes to neurodegeneration in Alzheimer's disease through aberrant cell cycle events.

Molecular neurodegeneration

Zhao F,Xu Y,Gao S,Qin L,Austria Q,Siedlak SL,Pajdzik K,Dai Q,He C,Wang W,O'Donnell JM,Tang B,Zhu X

Published figure using IBA1 polyclonal antibody (Product # PA5-21274) in Immunocytochemistry

Thu Sep 30 00:00:00 EDT 2021

METTL3-dependent RNA m⁶A dysregulation contributes to neurodegeneration in Alzheimer's disease through aberrant cell cycle events.

Molecular neurodegeneration

Zhao F,Xu Y,Gao S,Qin L,Austria Q,Siedlak SL,Pajdzik K,Dai Q,He C,Wang W,O'Donnell JM,Tang B,Zhu X

Published figure using IBA1 polyclonal antibody (Product # PA5-21274) in Immunocytochemistry

Thu Sep 30 00:00:00 EDT 2021

VPS35 D620N knockin mice recapitulate cardinal features of Parkinson's disease.

Aging cell

Niu M,Zhao F,Bondelid K,Siedlak SL,Torres S,Fujioka H,Wang W,Liu J,Zhu X

Published figure using IBA1 polyclonal antibody (Product # PA5-21274) in Immunohistochemistry

Sat May 01 00:00:00 EDT 2021

Profound downregulation of neural transcription factor Npas4 and Nr4a family in fetal mice neurons infected with Zika virus.

PLoS neglected tropical diseases

Alpuche-Lazcano SP,Saliba J,Costa VV,Campolina-Silva GH,Marim FM,Ribeiro LS,Blank V,Mouland AJ,Teixeira MM,Gatignol A

PA5-21274 was used in Western Blotting to identify a profound dysregulation of neural transcription factors with an overexpression of miR-7013-5p that results in decreased Nr4a3 expression, likely a main contributor to ZIKV-induced neuronal dysfunction.

Sat May 01 00:00:00 EDT 2021

Complement mediates neuroinflammation and cognitive decline at extended chronic time points after traumatic brain injury.

Acta neuropathologica communications

Mallah K,Couch C,Alshareef M,Borucki D,Yang X,Alawieh A,Tomlinson S

Published figure using IBA1 polyclonal antibody (Product # PA5-21274) in Immunohistochemistry

Tue Apr 20 00:00:00 EDT 2021

Sensory Ganglia-Specific TNF Expression Is Associated With Persistent Nociception After Resolution of Inflammation.

Frontiers in immunology

Gonçalves WA,Rezende BM,de Oliveira MPE,Ribeiro LS,Fattori V,da Silva WN,Prazeres PHDM,Queiroz-Junior CM,Santana KTO,Costa WC,Beltrami VA,Costa VV,Birbrair A,Verri WA,Lopes F,Cunha TM,Teixeira MM,Amaral FA,Pinho V

Published figure using IBA1 polyclonal antibody (Product # PA5-21274) in Immunofluorescence

Mon Nov 16 00:00:00 EST 2020

Bilateral Implantation of Shear Stress Modifier in ApoE Knockout Mouse Induces Cognitive Impairment and Tau Abnormalities.

Frontiers in aging neuroscience

Nie S,Tan Y,Zhang Z,Chen G,Xiong J,Hu D,Ye K,Zhang Y,Cao X,Chen L,Zhang Z

PA5-21274 was used in Immunohistochemistry to investigate the mechanism underlying vascular cognitive impairment in an Apolipoprotein E knockout mouse model fed with western style food with implantation of bilateral shear stress modifiers.

Tue Sep 29 00:00:00 EDT 2020

AAV cis-regulatory sequences are correlated with ocular toxicity.

Proceedings of the National Academy of Sciences of the United States of America

Xiong W,Wu DM,Xue Y,Wang SK,Chung MJ,Ji X,Rana P,Zhao SR,Mai S,Cepko CL

PA5-21274 was used in Immunohistochemistry to suggest that ocular AAV toxicity is associated with certain AAV cis-regulatory sequences and/or their activity and that retinal damage occurs due to responses by the RPE and/or microglia.

Tue Mar 19 00:00:00 EDT 2019

Crocin Inhibits Oxidative Stress and Pro-inflammatory Response of Microglial Cells Associated with Diabetic Retinopathy Through the Activation of PI3K/Akt Signaling Pathway.

Journal of molecular neuroscience : MN

Yang X,Huo F,Liu B,Liu J,Chen T,Li J,Zhu Z,Lv B

PA521274 was used in western blot to assess the role of microglial cells in diabetic retinopathy

Sat Apr 01 00:00:00 EDT 2017

Bioinformatics

Protein Aliases: AIF-1; Allograft inflammatory factor 1; balloon angioplasty responsive transcript; balloon angioplasty-responsive transcript 1; DADB-70P7.8; DASS-82G15.2; Em:AF129756.17; Iba; interferon gamma responsive transcript; ionized calcium binding adapter molecule 1; Ionized calcium-binding adapter molecule 1; Microglia response factor; MRF-1; protein BART-1; Protein G1; testis specific

Gene Aliases: AI607846; AIF-1; AIF1; BART-1; Bart1; D17H6S50E; G1; IBA1; IRT-1; IRT1; mrf-1; Mrf1

UniProt ID: (Human) P55008, (Mouse) O70200, (Rat) P55009

Entrez Gene ID: (Human) 199, (Mouse) 11629, (Rat) 29427

Molecular Function: calmodulin-related